绵羊肺腺瘤病相关基因突变的检测及绵羊、山羊内源性反转录病毒的扩增
|
摘要
研究背景：绵羊肺腺瘤病(ovine pulmonary adenomatosis, OPA)是一种由绵羊肺腺瘤病病毒(jaagsiekte sheep retrovirus, JSRV)引起的肺脏肿瘤性疾病。病原JSRV为β反转录病毒。自然感染绵羊潜伏期为2～24个月,人工感染绵羊潜伏期为3-7个月,患病绵羊病死率为100%。世界动物卫生组织将该病列为B类传染病。JSRV的Env蛋白是在体内和体外起主要作用的癌蛋白,有研究表明PI3K/AKT和RAS-MEK-MAPK是JSRV Env蛋白转化细胞的信号通路。建立特异、快速诊断方法是本病防控的基础。病毒致病机制的研究是深入了解本病的重点。
研究目的方法：建立特异性诊断OPA的方法和研究OPA与癌症相关基因突变的关系。本实验在JSRV结构基因研究的基础上参照GenBank中登录的JSRV基因序列,针对编码Env蛋白YXXM基序列设计了特异性PCR引物,建立了PCR检测方法,并对保存的病料进行了检测。参照SANGER、UCSC、GenBank数据库对nras、kras、pik3ca、egfr,p53基因发生突变较高的位点的外显子进行扩增。应用SSCP-PCR技术对扩增产物进行分析,对疑似突变的扩增产物进行测序检测。参照GenBank中登录的enJSRV基因序列,设计三对特异性引物扩增出一株绵羊内源性肺腺瘤反转录病毒和一株山羊内源性肺腺瘤反转录病毒基因片段。以山羊基因组DNA为模板,应用PCR技术扩增山羊内源性病毒基因片段,分别连接PMD-18T载体并测序。利用Methyl Primer Express v1.0查找绵羊和山羊enJSRV启动子区CpG岛,各设计一对甲基化引物和一对非甲基化引物。以处理的绵羊和山羊基因组DNA为模板,检测绵羊胎儿不同组织基因组DNA的甲基化情况,和不同山羊组织基因组DNA甲基化情况。构建含enJSRV的pro、env融合基因的真核表达载体EGFP-enpro、pcDNA3.1-enenv并在小鼠肺上皮细胞系TC-1中表达。
实验结果：(1)建立的特异性PCR检测方法,具有高的特异性、重复性,可检测出最低10～20拷贝的JSRV前病毒DNA。检测出JSRV阳性样品10例。(2)应用PCR-SSCP检测法,对10例JSRV阳性样品和10例阴性样品进行检测,结果在2例阳性标本中发现kras基因第二外显子第12氨基酸杂合型突变,CDS突变356)T,氨基酸突变G12V。1例阳性样品检测出p53基因第7外显子杂合型突变,CDS突变825T>C,氨基酸未发生突变C275C。(3)完成了一株绵羊内源性肺腺瘤病毒和一株山羊内源性肺腺瘤病毒基因序列的测定和序列分析,结果显示测得的绵羊enJSRVN序列与enJSRV-20(EF680302.1)病毒株的核苷酸同源性为99.4%,与外源性绵羊肺腺瘤病毒JS7(AF357971.1)病毒株的核苷酸同源性为90.9%。测得的山羊GERV序列与内源性绵羊肺腺瘤病毒株enJSRV-5(EF680305.1)的核苷酸同源性为94.2%,与外源性绵羊肺腺瘤病毒株JS-7(AF357971.1)的核苷酸同源性为88.1.%。扩增的绵羊和山羊内源性反转录病毒的同源性为93.1%。对enJSRV基因启动子甲基化检测,结果显示绵羊和山羊不同组织中均存在甲基化与非甲基化修饰的病毒启动子。(4)经过酶切鉴定pro.eNV基因成功连接入真核表达载体pEGFP-C1和pcDNA3.1(+),蛋白免疫荧光和Western BlOt方法检测显示重组质粒、EGFP-enpro、 pcDNA3.1-enenv能够在真核细胞中瞬时表达.
结论：(1)本研究建立了JSRV的特异性检测方法,对OPA的早期诊断和防控具有实用价值。(2)检测到JSRV感染绵羊肿瘤组织基因组中kras.p53基因突变,对研究OPA的致病机制具有重要参考价值。(3)成功扩增出一株绵羊和一株山羊enJSRV基因序列,检测了绵羊和山羊enJSRV启动子区的甲基化修饰情况。(4)分别构建了包含enJSRV的pro、env基因的真核表达质粒,为研究enJSRV的功能和抑制JSRV感染的作用奠定了基础。
BACKGROUND:Jaagsiekte sheep retrovirus (JSRV) is a type βretrovirus specifically associated with a contagious lung tumor of sheep, ovine pulmonary adenomatosis (OPA).Incubation period of OPA cause in nature is2～24months, incubation period of OPA cause by experiment is3～7months. OPA is the B-type infectious disease according to the disese classification from OIE.The JSRV envelope glycoprotein (Env) functions as a dominant oncoprotein in vitro and in vivo. Several reports suggested that the phosphatidylinositol3-kinase/AKT pathway and RAS-MEK-MAPK pathway are important for transformation of cells. Establish specific diagnosis method is the foundation to control and prevention the disease. mechanism of JSRV is key point to understand the disease.
OBJECTIVE AND METHOD:Jaagsiekte sheep retrovirus should be detected by using specific PCR and invested the relationship between OPA and gene mutation. The specific primers were designed according to sequences of JSRV in GenBank. And the peptide sequence of env, YXXM was take into count in designing of primers. The spefic PCR method could used in detection of JSRV,and the clinical sambles were detection by the method. High mutation rate site in gene nras、kras、pik3ca、egfr、p53was confirmed by search SANGER data bank.Exon of gene nras、kras、pik3ca、egfr、p53was confirmed by search data bank of UCSC and GenBank. Gene mutation site was detection by using technique of PCR-SSCP and the samples were sequencing. Three pairs of primers were designed according to the sequence of strain enJSRV-20. Sequences of sheep's and goat's enJSRV were amplified by PCR technology and these PCR products were cloned into pMD18-T vector and then sequenced.
RESULTS:(1)The method of spefic PCR showed high sensitivity, With high specificity and repeatability. The lowest level of10～20copies of the virus.DNA or cDNA can be detected by the PCR method.(2)10clinic samples were positive detected by the method.10positive samples and10negative samples were detection by the method of PCR-SSCP,The mutation was detection in the second exon of kras in2positive sample. The mutation site was in the12th genetic code, Which nucleotide G was mution to T,amino Glycine was mutivon to valine.The mutation of7th exon of gene p53was detectied in one sample. Which nucleotide T mution to C, amino was not mutivon.(3)The complete genome of goat's and sheep's enJSRV were obtained successfuly. Analysis results showed that the sequence of sheep's enJSRV was99.4%similarity with enJSRV-20and90.9%similarity with strain JS7. Analysis results show that the sequences of goat's enJSRV was94.2%similarity with enJSRV-5and88.1%similarity with strain JS7. The sequence of sheep'senJSRV was93.1%similarity with goat's enJSRV. enJSRV promoter methylation analysis of sheep and goat tissue showed that:methylation and unmethylation enJSRV promoter was detection in all the organization DNA.(4)The gene pro、env were link to eukaryotic expression vector confirm by the enzyme cut reaction, Protein immune fluorescence and Western Blot test showed that constructuring plasmid EGFP-enpro%pcDNA3.1-enenv can the instantaneous expression in eukaryotic cells.
CONCLUSIONS:(1)The specific RCR detection method of JSRV was established in this study, and it is important for studying OPA control measures.(2)Gene mutations of kras and p53was detection in JSRV infection tumor tissue will be useful in the molecular pathogenesis and further study of OPA.(3) The complete genome of goat'sand sheep's enJSRV were amplication and sequence analysis, methylation and unmethylation enJSRV promoter were detection in all the organization DNA.(4) The construction of the enJSRV pro and env eukaryotic express plasmid will provides an important experimental platform to study of enJSRV function and inhibiting virus infection.
研究目的方法：建立特异性诊断OPA的方法和研究OPA与癌症相关基因突变的关系。本实验在JSRV结构基因研究的基础上参照GenBank中登录的JSRV基因序列,针对编码Env蛋白YXXM基序列设计了特异性PCR引物,建立了PCR检测方法,并对保存的病料进行了检测。参照SANGER、UCSC、GenBank数据库对nras、kras、pik3ca、egfr,p53基因发生突变较高的位点的外显子进行扩增。应用SSCP-PCR技术对扩增产物进行分析,对疑似突变的扩增产物进行测序检测。参照GenBank中登录的enJSRV基因序列,设计三对特异性引物扩增出一株绵羊内源性肺腺瘤反转录病毒和一株山羊内源性肺腺瘤反转录病毒基因片段。以山羊基因组DNA为模板,应用PCR技术扩增山羊内源性病毒基因片段,分别连接PMD-18T载体并测序。利用Methyl Primer Express v1.0查找绵羊和山羊enJSRV启动子区CpG岛,各设计一对甲基化引物和一对非甲基化引物。以处理的绵羊和山羊基因组DNA为模板,检测绵羊胎儿不同组织基因组DNA的甲基化情况,和不同山羊组织基因组DNA甲基化情况。构建含enJSRV的pro、env融合基因的真核表达载体EGFP-enpro、pcDNA3.1-enenv并在小鼠肺上皮细胞系TC-1中表达。
实验结果：(1)建立的特异性PCR检测方法,具有高的特异性、重复性,可检测出最低10～20拷贝的JSRV前病毒DNA。检测出JSRV阳性样品10例。(2)应用PCR-SSCP检测法,对10例JSRV阳性样品和10例阴性样品进行检测,结果在2例阳性标本中发现kras基因第二外显子第12氨基酸杂合型突变,CDS突变356)T,氨基酸突变G12V。1例阳性样品检测出p53基因第7外显子杂合型突变,CDS突变825T>C,氨基酸未发生突变C275C。(3)完成了一株绵羊内源性肺腺瘤病毒和一株山羊内源性肺腺瘤病毒基因序列的测定和序列分析,结果显示测得的绵羊enJSRVN序列与enJSRV-20(EF680302.1)病毒株的核苷酸同源性为99.4%,与外源性绵羊肺腺瘤病毒JS7(AF357971.1)病毒株的核苷酸同源性为90.9%。测得的山羊GERV序列与内源性绵羊肺腺瘤病毒株enJSRV-5(EF680305.1)的核苷酸同源性为94.2%,与外源性绵羊肺腺瘤病毒株JS-7(AF357971.1)的核苷酸同源性为88.1.%。扩增的绵羊和山羊内源性反转录病毒的同源性为93.1%。对enJSRV基因启动子甲基化检测,结果显示绵羊和山羊不同组织中均存在甲基化与非甲基化修饰的病毒启动子。(4)经过酶切鉴定pro.eNV基因成功连接入真核表达载体pEGFP-C1和pcDNA3.1(+),蛋白免疫荧光和Western BlOt方法检测显示重组质粒、EGFP-enpro、 pcDNA3.1-enenv能够在真核细胞中瞬时表达.
结论：(1)本研究建立了JSRV的特异性检测方法,对OPA的早期诊断和防控具有实用价值。(2)检测到JSRV感染绵羊肿瘤组织基因组中kras.p53基因突变,对研究OPA的致病机制具有重要参考价值。(3)成功扩增出一株绵羊和一株山羊enJSRV基因序列,检测了绵羊和山羊enJSRV启动子区的甲基化修饰情况。(4)分别构建了包含enJSRV的pro、env基因的真核表达质粒,为研究enJSRV的功能和抑制JSRV感染的作用奠定了基础。
BACKGROUND:Jaagsiekte sheep retrovirus (JSRV) is a type βretrovirus specifically associated with a contagious lung tumor of sheep, ovine pulmonary adenomatosis (OPA).Incubation period of OPA cause in nature is2～24months, incubation period of OPA cause by experiment is3～7months. OPA is the B-type infectious disease according to the disese classification from OIE.The JSRV envelope glycoprotein (Env) functions as a dominant oncoprotein in vitro and in vivo. Several reports suggested that the phosphatidylinositol3-kinase/AKT pathway and RAS-MEK-MAPK pathway are important for transformation of cells. Establish specific diagnosis method is the foundation to control and prevention the disease. mechanism of JSRV is key point to understand the disease.
OBJECTIVE AND METHOD:Jaagsiekte sheep retrovirus should be detected by using specific PCR and invested the relationship between OPA and gene mutation. The specific primers were designed according to sequences of JSRV in GenBank. And the peptide sequence of env, YXXM was take into count in designing of primers. The spefic PCR method could used in detection of JSRV,and the clinical sambles were detection by the method. High mutation rate site in gene nras、kras、pik3ca、egfr、p53was confirmed by search SANGER data bank.Exon of gene nras、kras、pik3ca、egfr、p53was confirmed by search data bank of UCSC and GenBank. Gene mutation site was detection by using technique of PCR-SSCP and the samples were sequencing. Three pairs of primers were designed according to the sequence of strain enJSRV-20. Sequences of sheep's and goat's enJSRV were amplified by PCR technology and these PCR products were cloned into pMD18-T vector and then sequenced.
RESULTS:(1)The method of spefic PCR showed high sensitivity, With high specificity and repeatability. The lowest level of10～20copies of the virus.DNA or cDNA can be detected by the PCR method.(2)10clinic samples were positive detected by the method.10positive samples and10negative samples were detection by the method of PCR-SSCP,The mutation was detection in the second exon of kras in2positive sample. The mutation site was in the12th genetic code, Which nucleotide G was mution to T,amino Glycine was mutivon to valine.The mutation of7th exon of gene p53was detectied in one sample. Which nucleotide T mution to C, amino was not mutivon.(3)The complete genome of goat's and sheep's enJSRV were obtained successfuly. Analysis results showed that the sequence of sheep's enJSRV was99.4%similarity with enJSRV-20and90.9%similarity with strain JS7. Analysis results show that the sequences of goat's enJSRV was94.2%similarity with enJSRV-5and88.1%similarity with strain JS7. The sequence of sheep'senJSRV was93.1%similarity with goat's enJSRV. enJSRV promoter methylation analysis of sheep and goat tissue showed that:methylation and unmethylation enJSRV promoter was detection in all the organization DNA.(4)The gene pro、env were link to eukaryotic expression vector confirm by the enzyme cut reaction, Protein immune fluorescence and Western Blot test showed that constructuring plasmid EGFP-enpro%pcDNA3.1-enenv can the instantaneous expression in eukaryotic cells.
CONCLUSIONS:(1)The specific RCR detection method of JSRV was established in this study, and it is important for studying OPA control measures.(2)Gene mutations of kras and p53was detection in JSRV infection tumor tissue will be useful in the molecular pathogenesis and further study of OPA.(3) The complete genome of goat'sand sheep's enJSRV were amplication and sequence analysis, methylation and unmethylation enJSRV promoter were detection in all the organization DNA.(4) The construction of the enJSRV pro and env eukaryotic express plasmid will provides an important experimental platform to study of enJSRV function and inhibiting virus infection.
引文
1 马学恩.家畜病理学[M].北京：中国农业出版社,2007,385-386
2 殷震,刘景华.动物病毒学.[M]第二版,北京：科学出版社,1997,836-894
3 邓普辉.绵羊肺腺瘤病一人细支气管-肺泡细胞癌的类似物.中国兽医科技,1988(2)：60-61
4 邓普辉.新疆绵羊肺腺瘤病调查[J].中国兽医杂志,1981(11)：4-5
5 幺宏强,其木格,马学恩.绵羊肺腺瘤病自然病例肺组织的超微结构观察[J].电子显微学报,2006：240
6 Grego E, et al. Colostrum and milk can transmit jaagsiekte retrovirus to lambs [J]. Vet Microbiol,2008,130 (3-4):247-257
7 Voigt K,et al. Eradication of ovine pulmonary adenocarcinoma by motherless rearing of lambs[J]. Vet Rec,2007,161 (4):129-132
8 Salvatori D,et al. Successful induction of ovine pulmonary adenocarcinoma in lamb of different ages and detection of viraemia during the preclinical period[J]. Gen Virol,2004.85(11):3319-3324
9 么宏强.绵羊肺腺瘤病毒人工感染羔羊淋巴细胞动态变化的研究[J].黑龙江畜牧兽医,2007(12)：69-71
10 Cousens C, et al. Evaluation of the efficacy of clinical diagnosis of ovine pulmonary adenocarcinoma[J]. Vet Rec,2008,162(3):88-90
11 Oda SS,SA Youssef. Immunohistochemical and histopathological findings of ovine pulmonary adenocarcinoma(Jaagsiekte) in Egyptian sheep[J].Trop Anim Health Prod, 2011,43(8):1611-1615
12 De Las HM, et al.A PCR technique for the detection of Jaagsiekte sheep retrovirus in the blood suitable for the screening of ovine pulmonary adenocarcinoma in field conditions [J].Res Vet Sci,2005,79 (3):259-264
13 Caporale M, et al.Expression of the jaagsiekte sheep retrovirus envelopegly sheep coprotein is sufficient to induce lung tumors in [J]. Virol,2006,80(16):8030-8037
14 Chitra E, et al. Functional interaction between env oncogene from Jagsiekte sheep retrovirus and tumor suppressor Sprouty2[J]. Retrovirology,2010(7):62
15 Palmarini M, et al. Jaagsiekte sheep retrovirus is necessary and sufficient toinduce a contagious lung cancer in sheep[J]. Virol,1999,73(8):6964-6972
16 Wootton SK,CL Halbert, AD Miller. Sheep retrovirus structural protein induceslung tumours[J]. Nature,2005,434 (7035):904-907
17 Miyazawa T.Receptors for animal retroviruses[J]. Uirusu,2009,59 (2):223-242
18 Vigdorovich V, RK Strong,AD Miller. Expression and characterization of a soluble, active form of the jaagsiekte sheep retrovirus receptor, Hya12[J]. Virol,2005, 79(1):79-86
19 Miller AD. Hyaluronidase 2 and its intriguing role as a cell-entry receptor for oncogenic sheep retroviruses [J].Semin Cancer Biol,2008,18 (4):296-301
20 Garcia-GotiM, et al.Sheep pulmonary adenomatosis:characterization of two patholo-gical forms associated with aagsiekte retrovirus [J]. Comp pthol,2000,122 (1):55-65
21 Maeda N, et al. Roles of the RAS-MEK-mi togen-act ivated protein kinaseand phosphate-idylinositol 3-kinase-AKT-mTOR pathways in Jaagsiekte sheep retrovirus-induced transformation of rodent fibroblast and epithelial cell lines[J]. Virol,2005, 79(7):4440-4450
22 Palmarini M, et al.A phosphat idylinositol 3-kinase docking site in the cytoplasmic tail of the Jaagsiekte sheep retrovirus transmembrane protein is essential for envelope induced transformation of NIH 3T3 cells[J]. Virol,2001,75 (22):11002-11009
23 De Las HM, et al. In-situ demonstration of mitogen-activated protein kinase Erk 1/2 signalling pathway in contagious respiratory tumours of sheep and goats[J]. Comp Pathol,2006,135(1):1-10
24 Liu SL, MI Lerman,AD Miller, Putative phosphatidylinositol 3-kinase (PI3K) binding motifs in ovine betaret rovirus env proteins are not essential for rodent fibroblast transformation and PI3K-AKT activation[J]. Virol,2003,77(14):7924-7935
25 Herring AJ,et al. Further evidence for a retrovirus as the aetiological agent of sheep pulmonary adenomatosis (jaagsiekte) [J]. Vet Microbiol,1983,8 (3):237-249
26 邓普辉,简子健,白惠敏.新疆绵羊肺腺瘤病回顾性研究[J].中国兽医杂志,1996(1)：3-5
27 林曦,郝先谱,赵振华.内蒙古动物肿瘤的调查[J].中国兽医病理学研究会肿瘤病理学专题讨论论文集,1985
28 Wander a JG. Pneumonia in sheep in Kenia. Part I Bacterial and parasitic pneumonia [J]. Bulletin of epizootic disease of Africa.1967.15:245-248
29 ustin RC.Ovine Jaagsiekte.Journal of the South African[J]. Veterinary Medical Association.1969,40:3-23
30 Sharp JM,Angus KW,Gray EW Scott. FMM. Rapid transmission of sheep pulmonary Adenomatosis(jaasiekte) in young lambs[J].Archives of Virology.1983,78:89-95
31 Niels Dungal.Experiments with Jaagsiekte[J]. Am J Pathol.1946,22 (4):737-759
32 Smith W,Mackay JMK. Morphological observations on a virus associated With sheep pulmonary adenomatosis(jaagsiekte) [J]. Journal of Comparative Pathology.1969,79: 421-425
33 Wandera JG. Sheep pulmonary adenomatosis (Jaagsiekte) [J].Advances in Veterinary cience and ComparativeMedicine.1971,15:251-283
34 Perk K, Hod I, Presentey B, et al.Lung carcinoma of sheep(jaagsiekte)[J].Journal of the National Cancer Institute.1971,47:197-205
35 Martin WB, Scott FMM,Sharp JM,Angus KW. Experimental production of sheep pulmonary adenomatosis (Jaagsiekte) [J].Nature.1976,264:183-184
36 Sharp JM, Herr ing AJ. Sheep pulmonary adenomatosis:demonstration of a protein which cross-reacts with the major core proteins of Mason-Pfizer monkey virus and mouse mammary tumour virus[J]. Gen Virol.1983,64:2323-2327
37 Myer MS, York DF,et al.The possible involvementof immunosuppression caused by a lentivirusin the aetiology of jaagsiekte and pasteurellosis in sheep [J]. Onderstepoort J Vet Res.1988,55:127-133
38 Verwoerd DW. Jaagsiekte (ovine pulmonary adenomatosis)virus. In:Dinta Z and Morein B(eds)[J]. Virus infections of ruminants.Elsevier,453-462
39 Afonso pv,et al. Centrosome and retroviruses:the dangerous liaisons [J]. Retrovirology,2007,4:27
40 Hecht SJ.et al.Distribution of endogenous type B and type D sheep retrovirus sequences in ungulates and other mammals[J]. Proc Natl Acad SciUSA,1996,93(8): 3297-3302
41 York F,Vigne R,Verwoerd DW, Querat G.Nucleotide sequence of the jaagsiekte retrovirus, an exogenous and endogenous type D and B retrovirus of sheep and goats [J]. Virol.1992,66 (8):4930-4939
42 Demartini JC, York D.Retrovirus-associated neoplasms of the respiratory system of sheep and goatsOvine pulmonary carcinoma and enzoot ic nasal tumor [J]. Vet Clin North Am Food Anim Pract.1997,13:55-70
43 中华人民共和国农业部公告动物疫病病种名录[M].《农业科技通讯》2000.10
44 Coil DA, et al. Jaagsiekte sheep retrovirus env protein stabilizes retrovirus vectors against inactivation by lung surfactant, centrifugation,and freeze-thaw cycling[J]. Virol,2001,75 (18):8864-8868
45 Cousens C, Thonur L, ImLach S. lung fluid produced by ovine pulmonary adenocarcinoma affected sheep and can survive for several weeks at ambient temperatures [J]. Res Vet Sci,2009,87(1):154-156
46 Summers C, et al. Jaagsiekte sheep retrovirus-specific immune responses induced by vaccination:a comparison of immunisation strategies [J]. Vaccine,2006,24(11): 1821-1829
47 Ortin A, et al.Lack of a specific immune response against a recombinant capsid protein of Jaagsiekte sheep retrovirus in sheep and goats naturally affected by enzootic nasal tumour or sheep pulmonary adenomatosis [J]. Vet Immunol Immunopathol, 1998,61(2-4):229-237
48 Hudachek SF,et al.Lung tumor development and spontaneous regression in lambs coinfected with Jaagsiekte sheep retrovirus and ovine lentivirus[J]. Vet Pathol, 2010,47(1):148-162
49 Cutlip RC,Young S, et al. Sheep pulmonary adenomatosis (jaagsiekte)in the United States [J]. American Journal of Veterinary Research.1982,43:2108-2113
50 Scott FMM, Sharp JM, Angus KW. Infection of specific-pathogen free lambs with a herpesvirus isolated from pulmonary adenomatosis [J].Archivo Virology.1984,80: 147-162
51 Holland MJ,et al.Jaagsiekte retrovirus is widely distributed both in T and B lymphocytes and in mononuclear phagocytes of sheep with naturally and experimentally acquired pulmonary adenomatosis [J].Virol,1999,73(5):4004-4008
52 Beytut EM,et al.Sozmen S Erginsoy. Immunohistochemical detection of pulmonary surfactant proteins and retroviral antigens in the lungs of sheep with pulmonary adenomatosis [J].Comp Pathol,2009,140(1):43-53
53 刘霄卉.绵羊肺腺瘤病毒囊膜基因表面蛋白的表达及其单克隆抗体的制备[J].中国预防兽医学报,2008(6)：469-472
54 Lewis FI, et al. Diagnostic accuracy of PCR for Jaagsiekte sheep retrovirus using field data from 125 Scottish sheep flocks[J].Vet J,2011,187 (1):104-108
55 Luo JR, et al. A specific PCR assay and a nested PCR assay for the screening of ovine pulmonary adenomatosis [J]. Bing Du Xue Bao,2009,25 (2):125-130
56 Voigt K, et al.PCR examination of bronchoalveolar lavage samples is a useful tool in pre-clinical diagnosis of ovine pulmonary adenocarcinoma (Jaagsiekte)[J].Res. Vet Sci,2007,83(3):419-427
57 Cousens C, et al.PCR-based detection and partial characterization of a retrovirus associated with contagious intranasal tumors of sheep and goats[J]. Virol,1996, 70(11):7580-7583
58 Gonzalez L, et al. Jaagsiekte sheep retrovirus can be detected in the peripheral blood during the pre-clinical period of sheep pulmonary adenomatosis [J]. Gen Virol, 2001,82(6):1355-1358
59 Sanna MP, et al. Association of jaagsiekte sheep retrovirus with pulmonary carcinoma in Sardinian moufflon (Ovis musimon) [J]. Comp Pathol,2001,125(2-3):145-152
60 Morozov VA,et al.Detection and characterization of betaretroviral sequences, related to sheep Jaagsiekte virus, in Africans from Nigeria and Cameroon[J]. Virology,2004,327 (2):162-168
61 Fox KA,et al.Paranasal sinus masses of Rocky Mountain bighorn sheep (Ovis canadensis Canadensis)[J]. Vet Pathol,2011,48(3):706-712
62 Maeda N, et al. Surveillance of Jaagsiekte sheep retrovirus in sheep in Hokkaido, the northern island of Japan[J].Vet Med Sci,2011,73(11):1493-1495
63 Suau F, et al. TelomeRASe activation in a model of lung adenocarcinoma. Eur Respir [J]. 2006,27(6):1175-1182
64 Palmarini M, H Fan, JM Sharp. Sheep pulmonary adenomatosis:a unique model of retrovirus associated lung cancer [J].Trends Microbiol,1997,5(12):478-483
65 Palmarini M, H Fan. Retrovirus-induced ovine pulmonary adenocarcinoma,an animal model for lung cancer[J].Natl Cancer Inst,2001,93 (21):1603-1614
66 Occa S, et al. Presence of Jaagsiekte sheep retrovirus in tissue sections from human bronchioloalveolar carcinoma depends on patients'geographical origin[J].Hum Pathol,2008,39 (2):303-304
67 Mornex JF,et al.Pathology of human bronchioloalveolar carcinoma and its relationship to the ovine disease[J].Curr Top Microbiol Immunol,2003,275:225-248
68 Hurnann-Ziehank E, et al. Ovine pulmonary adenocarcinoma as an animal model of progressive lung cancer and the impact of nutritional selenium supply [J]. Trace Elem Med Biol,2011,25 (11):30-34
69 Murgia C, et al.Lung adenocarcinoma originates from retrovirus infection of proferating type 3 pneumocytes during pulmonary post-natal development or tissue repair[J].PLoS Pathog,2011,7(3):100-114
70 Hiatt KM, WE Highsmith. Lack of DNA evidence for jaagsiekte sheep retrovirus in human bronchioloalveolar carcinoma[J]. Hum Pathol,2002,33 (6):680
71 Yousem SA,et al.Absence of jaagsiekte sheep retrovirus DNA and RNA in bronchioloalveolar and conventional human pulmonary adenocarcinoma by PCR and RT-PCR analysis[J]. Hum Pathol,2001,32 (10):1039-1042
72 Hopwood P, et al. Absence of markers of betaretrovirus infection in human pulmonary adenocarcinoma [J]. Hum Pathol,2010,41 (11):1631-1640
73 DeMartini JC,et al.Jaagsiekte sheep retrovirus proviral clone JSRV(JS7), derived from the JS7 lung tumor cell line, induces ovine pulmonary carcinoma and is integrated into the surfactant protein A gene[J]. Virol,2001,75(9):4239-4246
74 Ni tta T, et al. Identification and mutational analysis of a Rej response element in jaagsiekte sheep retrovorus RNA[J]. Virol,2009,83(23):12499-12511
75 Chitra E, Yu SL, Hsiao KN,Shao HY. Generation and characterization of JSRV envelope transgenic mice in FVB background[J]. Virology.2009,393(1):120-126
76 Ortin A, et al. Characterization of enzootic nasal tumour virus of goats:complete sequence and tissue distribution[J].Gen Virol,2003.84:2245-2252
77 Cousens C, et al. Complete sequence of enzootic nasal tumor virus,a retrovirus associated with transmissible intranasal tumors of sheep[J]. Virol,1999,73(5): 3986-3993
78 Kaufmann C, et al. Enzootic nasal adenocarcinoma in a dairy sheep flock [J]. Schweiz Arch TierheilkD,2008,150(6):297-302
79 De Las, et al. Enzootic nasal adenocarcinoma of sheep and goats [J]. Curr Top Microbiol Immunol,2003,275,201-223
80 Wootton SK.CLHalbert, AD Miller. envelope proteins of Jaagsiekte sheep retrovirus and enzootic nasal tumor virus induce similar bronchioalveolar tumors in lungs of mice[J]. Virol,2006,80(18):9322-9325
81 Dunlap KA,M Palmarini, TE Spencer, Ovine endogenous betaretroviruses (enJSRV) and placental morphogenesis. Placenta,2006,27:135-140
82 Carlson J, et al. Chromosomal distribution of endogenous Jaagsiekte sheep retrovirus proviral sequences in the sheep genome [J]. Virol,2003.77 (17):9662-9668
83 Hecht SJ,JO.Carlson,JC DeMartin. Analysis of a type D retroviral capsid gene expressed in ovine pulmonary carcinoma and present in both affected and unaffected sheep genomes[J]. Virology,1994,202(1):480-484
84 Mortuza GB,et al. Structure of the capsid amino-terminal domain from the betaretrovirus, Jaagsiekte sheep retrovirus[J].J Mol Biol,2009,386 (4):1179-1192
85 Johnson C, et al. Enhanced proliferation of primary rat type Ⅱ pneumocytes by Jaagsiekte sheep retrovirus envelope protein[J].Virology,2011,412 (2):349-355
86 Golovkina TV,Jaffe AB,Ross SR. Coexpression of exogenous and endogenous mouse mammary tumour virus RNA in vivo results in viral recombination and broadens the virus host range [J].J Virol,1994,68:5019-5026
87 Golovkina TV, Chervonsky A.Dudley JP, Ross SR. Transgenic mouse mammary tumor virus superantigen expression prevents viral infection[J]. Cell,1992,69:637-645
88 Berkowitz R, Fisher J.Goff SP. RNA packaging [J]. Curr Top Microbiol Immunol 1996,214: 177-218
89 Rosati S,et al.An accessory open reading frame (orf-x) of jaagsiekte sheep retrovirus is conserved between different virus isolates[J]. Virus Res,2000,66(1): 109-116
90 McGee-Estrada K, H Fan, Comparison of LTR enhancer elements in sheep beta retroviruses:insights into the basis for tissue-specific expression[J]. Virus Genes,2007,35(2):303-312
91 McGee-Estrada K,et al.A Moloney murine leukemia virus driven by the Jaagsiekte sheep retrovirus enhancers shows enhanced specificity for infectivity in lung epithelial cells[J]. Virus Genes,2005,31 (3):257-263
92 McGee-Estrada KM, Palmarini H Fan. HNF-3beta is a critical factor for the expression of the Jaagsiekte sheep retrovirus long terminal repeat in type II pneumocytes but not in Clara cells[J]. Virology 2002,292(1):87-97
93 Sinn PL, et al. Inclusion of Jaagsiekte sheep retrovirus proviral elements markedly increases lentivirus vector pseudotyping efficiency [J]. Mol Ther,2005,11 (3):460-469
94 McGee-Estrada K, H Fan. In vivo and in vitro analysis of factor binding sites in Jaagsiekte sheep retrovirus long terminal repeat enhancer sequences:roles of HNF-3, NF-I, and C/EBP for activity in lung epithelial cells[J]. Virol,2006,80(1):332-341
95 Dakessian RM, H Fan. Specific in vivo expression in type II pneumocytes of the Jaagsiekte sheep retrovirus long terminal repeat in transgenic mice[J]. Virology, 2008,372(2):398-408
96 Palmarini M,et al.The long terminal repeat of Jaagsiekte sheep retrovirus is preferentially active in differentiated epithelial cells of the lungs [J]. Virol, 2000,74(13):5776-5787
97 Leroux C,et al.Jaagsiekte Sheep Retrovirus (JSRV):from virus to lung cancer in sheep [J]. Vet Res,2007,38 (2):211-228
98 Philbey AW,et al. Multiclonal pattern of Jaagsiekte sheep retrovirus integration sites in ovine pulmonary adenocarcinoma [J]. Virus Res,2006,117 (2):254-263
99 Cousens C,et al. Analysis of integration sites of Jaagsiekte sheep retrovirus in ovine pulmonary adenocarcinoma[J]. J Virol,2004.78 (16):8506-8512
100 Rai SK,Miller AD. Retrovirus vector bearing jaagsiekte sheep retrovirus env transducer human cells by using a new receptor localized to chromosome 3p21.3 [J]. Virol 2000,74:4698-4704
101 Vigdorovich V, AD Miller. Ability of hyaluronidase2 to degrade extracellular hyaluronan is not required for its function as a receptor for jaagsiekte sheep retrovirus [J].Virol,2007,81(7):3124-4129
102 Duh FM, et al. Amino acid residues that are important for Hya12 function as a receptor for jaagsiekte sheep retrovirus [J].Retrovirology,2005,2:59
103 Miller AD.Identification of Hya12 as the cell-surface receptor for jaagsiekte sheep retrovirus and ovine nasal adenocarcinoma virus [J]. CurrTop Microbiol Immunol, 2003,275:179-199
104 Van Hoeven NS, AD Miller. Improved enzootic nasal tumor virus pseudotype packaging cell lines reveal virus entry requirements in addition to the primary receptor Hya12[J]. Virol,2005,79(1):87-94
105 Rai SK, JC DeMartini.AD Miller. Retrovirus vectors bearing jaagsiekte sheep retrovirus env transduce human cells by using a new receptor localized to chromosome 3p21.3[J].Virol,2000,74(10):4698-4704
106 Liu SL, et al.Role of virus receptor Hya12 in oncogenic transformation of rodent fibroblasts by sheep betaretrovirus env proteins[J], Virol,2003,77(5):2850-2858
107 Sharath K Rai,Fuh-Mei Duh, Vladimir Vigdorovich,et al. Candidate tumor suppressor HYAL2 is a glycosylphosphatidylinositol (GPI)-anchored cell-surface receptor for jaagsiekte sheep retrovirus, the envelope protein of which mediates oncogenic transformation[J].2001,98 (8):4443-4448
108 Palmarini M.M Mura.TE Spencer. Endogenous betaretroviruses of sheep:teaching new lessons in retroviral interference and adaptation[J].J Gen Virol,2004,85 (Ptl): 1-13
109 Sanna E, et al. Endogenous jaagsiekte sheep retrovirus RNA is expressed by different cell types in ovine foetus and placenta[J]. Eur J Histochem,2002,46(3):273-280
110 Dunlap KA,et al.Endogenous retroviruses regulate periimplantation placental growth and differentiation[J]. Proc Natl Acad Sci USA,2006.103(39):14390-14395
111 DeMartini JC. Endogenous retroviruses related to jaagsiekte sheep retrovirus [J]. Curr Top Microbiol Immunol,2003,275:117-137
112 Spencer TE,M Palmarini. Endogenous retroviruses of sheep:a model system for understanding physiological adaptation to an evolving ruminant genome [J]. J Reprod Dev,2012.58(1):33-37
113 Palmarini M,et al.Molecular cloning and functional analysis of three type D endogenous retroviruses of sheep reveal a different cell tropism from that of the highly related exogenous jaagsiekte sheep retrovirus [J]. Virol,2000,74 (17):8065-8076
114 Black SG.et al. Endogenous retroviruses in trophoblast differentiation and placental development [J]. Am J Reprod Immunol 2010,64(4):255-264
115 Arnaud F,et al. Coevolution of endogenous betaretroviruses of sheep and their host[J].Cell Mol Life Sci,2008,65(21):3422-3432
116 Arnaud F,et al.A paradigm for virus-host coevolution:sequential counter-adaptations between endogenous and exogenous retroviruses [J].PLoS Pathog,2007, 3(11):e170
117 Frederick Arnaud.Marco Caporale,Mariana Varela. A Paradigm for Virus Host Coevolution:Sequential Counter-Adaptations between Endogenous and Exogenous Retroviruses [J]. PLoS Pathog,2007,3(11):e170
118 Kathrin A, Dunlap, Massimo Palmarini. Endogenous retroviruses regulate periimp-lantation placental growth and differentiation[J]. Proc Natl Acad Sci USA.2006, 103(39):165-176
119 Mura M, et al. Late viral interference induced by transdominant Gag of an endogenous retrovirus[J].Proc Natl Acad Sci USA,2004.101 (30):11117-11122
120 Arnaud F, PR Murcia,M Palmarini,Mechanisms of late restriction induced by an endogenous retrovirus [J]. Virol,2007,81 (20):11441-11451
121 Spencer TE, Mariana Varela, et al.Receptor usage and fetal expression of ovine endogenous betaret roviruses:implications for coevolution of endogenous and exogenous retroviruses[J]. Virol,2003,77 (1):749-753
122 Armezzani A, et al.The signal peptide of a recently integrated endogenous sheep betaretrovirus envelope plays a major role in eluding gag-mediated late restriction[J].Virol,2011,85 (14):7118-7128
123 Murcia PR,F Arnaud, M Palmarini, The transdominant endogenous retrovirus enJS56Al associates with and blocks intracellular trafficking of Jaagsiekte sheep retrovirus Gag[J]. Virol,2007,81 (4):1762-1772
124 Massimo Palmarini, Naoyoshi Maeda, Claudio Murgia, et al.A Phosphatidy linositol 3-Kinase Docking Site in the Cytoplasmic Tail of the Jaagsiekte Sheep Retrovirus Transmembrane Protein Is Essential for envelope-Induced Transformation of NIH 3T3 Cells[J]. Virol.2001,75 (22):11002-11009
125 Marceline C, Yi-Min Zheng, Lorraine M. Albritton, Fusogenicity of Jaagsiekte Sheep Retrovirus envelope Protein Is Dependent on Low pH and Is Enhanced by Cytoplasmic Tail Truncations [J].Virol.2008,82(5):2543-2554
126 Sharath K, Rai,Fuh-Mei Duh,Vladimir Vigdorovich,et al. Candidate tumor suppressor HYAL2 is a glycosylphosphatidylinositol (GPI)-anchored cell-surface receptor for jaagsiekte sheep retrovirus, the envelope protein of which mediates oncogenic transformation [J].Proc Natl Acad Sci USA,2001,98 (8):4443-4448
127 Miller AD,NS Van Hoeven,SL Liu. Transformation and scattering activities of the receptor tyrosine kinase RON/Stk in rodent fibroblasts and lack of regulation by the jaagsiekte sheep retrovirus receptor [J], BMC Cancer,2004,4:64
128 Danilkovitch-Miagkova A, et al. Hyaluronidase 2 negatively regulates RON receptor tyros ine kinase and mediates transformat ion of epithelial cells by jaagsiekte sheep retrovirus[J].Proc Natl Acad Sci USA,2003,100(8):4580-4585
129 Hull, S, H Fan.Mutational analysis of the cytoplasmic tail of jaagsiekte sheep retrovirus envelope protein[J]. Virol,2006,80(16):8069-8080
130 Massimo Palmarini,J Michael Sharp, Christine Lee. In Vitro Infection of Ovine Cell Lines by Jaagsiekte Sheep Retrovirus[J]. Virol,1999,73 (12):10070-10078
131 Hofacre A, H Fan. Multiple domains of the Jaagsiekte sheep retrovirus envelope protein are required for transformation of rodent fibroblasts[J]. Virol,2004,78: 10479-10489
132 Hallwirth C, et al. Variable regions 1 and 2 (VR1 and VR2) in JSRV gag are not responsible for the endogenous JSRV particle release defect[J]. Virus Genes,2005, 30(1):59-68
133 Naoyoshi Maeda,Wuxia Fu, Aurora Ortin. Roles of the RAS-MEK-Mitogen-Activated Protein Kinase and Phosphatidylinositol 3-Kinase-AKT-mTOR Pathways in Jaagsiekte Sheep Retrovirus-Induced Transformation of Rodent Fibroblast and Epithelial Cell Lines[J]. Virol,2005,79 (7):4440-4450
134 Mariana Varela, Matthew Golder, et al.A Large Animal Model to Evaluate the Effects of Hsp90 Inhibitors for the Treatment of Lung Adenocarcinoma [J]. Virology,2008, 371(1):206-215
135 Johnson C,K Sanders,H Fan. Jaagsiekte sheep retrovirus transformation in Madin Darby canine kidney epithelial cell three-dimensional culture[J].J Virol,2010, 84(10):5379-5390
136 Dani lkovi tch-Miagkova, AFM Duh, I Kuzmin, D Angeloni,SL Liu, AD Miller, MI Lerman. Hyaluronidase2 negatively regulates RON receptor tyrosine kinase and mediates transformation of epithelial cells by jaagsiekte sheep retrovirus[J]. Natl Acad Sci.USA 2003,100:4580-4585
137 Vaughan A E, et al.Lung cancer in mice induced by the jaagsiekte sheep retrovirus envelope protein is not maintained by rare cancer stem cells, but tumorigenicity does correlate with Wnt pathway activation[J]. Mol Cancer Res,2012,10 (1):86-95
138 Maeda N,et al.Transformation of mouse fibroblasts by Jaagsiekte sheep retrovirus envelope does not require phosphatidylinositol 3-kinase[J]. Virol,2003,77 (18): 9951-9959
139 Maeda N,et al Direct transformation of rodent fibroblasts by jaagsiekte sheep retrovirus DNA[J].Proc Natl Acad Sci USA,2001,98 (8):4449-4454
140 Shan-Lu Liu A, Dusty Miller. Transformation of Madin-Darby Canine Kidney Epithelial Cells by Sheep Retrovirus envelope Proteins [J].J Virol,2005,79 (2):927-933
141 Chetaille MC, S Olschwang, KRAS Mutation Spectrum Notably Diverges between Non-small Cell Lung and Colorectal Carcinomas[J].Thorac Oncol,2012.7 (4):773-774
142刘丽琴,张海燕,王捷KRAS基因突变检测与结直肠癌分子靶向治疗的研究进展[J].现代肿瘤医学,2011(4)：802-804
143 Jing Gao, Tingting Wang, Jing wei Yu, Yan-yan Li. Wild-Type KRAS and BRAF Could Predict Response to Cetuximab in Chinese Colorectal Cancer Patients[J]. Chinese Journal of Cancer Research,2011 (4):271-275
144 Ellis PM. KRAS Mutation Spectrum Notably Diverges between Non-small Cell Lung and Colorectal Carcinomas [J].Thorac Oncol,2012.7 (4):774
145隆艳艳,戴广海.胰腺癌中EGFR、KRAS及PI3K/AKT通路对EGFR-TKIs疗效预测的研究[J].军医进修学院学报,2011(11)：1185-1187
146 Buttitta F,et al.PIK3CA mutation and histological type in breast carcinoma:high frequency of mutations in lobular carcinoma [J]. Pathol,2006,208 (3):350-355
147 Kalinsky K,et al.PIK3CA mutation associates with improved outcome in breast cancer[J].Clin Cancer Res,2009,15(16):5049-5059
148 Cizkova M, et al. PIK3CA mutation impact on survival in breast cancer patients and in ERalpha, PR and ERBB2-based subgroups[J]. Breast Cancer Res,2012,14 (1):R28
149 Nosho K, et al. PIK3CA mutation in colorectal cancer:relationship with genetic and epigenetic alterations [J]Neoplasia,2008,10(6):534-541
150 Yamamoto S. PIK3CA mutation is an early event in the development of endometrios is associated ovarian clear cell adenocarcinoma [J]. Pathol,2011,225 (2):189-194
151 Kozaki K, et al.PIK3CA mutation is an oncogenic aberration at advanced stages of oral squamous cell carcinoma [J].Cancer Sci,2006,97 (12):1351-1358
152 Ogino S, et al. PIK3CA mutation is associated with poor prognosis among patients with curatively resected colon cancer[J]. Clin Oncol,2009.27 (9):1477-1484
153 Kato S,et al.PIK3CA mutation is predictive of poor survival in patients with colorectal cancer [J].Int J Cancer,2007,121(8):1771-1778
154 Thunnisse E, et al. EGFR-mutation in non-small cell lung carcinoma. TreaTment with tyrosine kinase inhibitors possible[J]. Ned Tijdschr GeneeskD,2011,155:A2554
155 Li F, et al.Role of EGFR mutation status in patients with stage Ⅲ non-squamous non-small cell lung cancer treated with chemoradiotherapy [J].Zhongguo Fei Ai Za Zhi,2011,14(9):715-718
156 Saitoh M,et al.An early event of EGFR mutation in pleomorphic carcinoma of the lung[J].Int J Clin Oncol,2011,16 (6):770-773
157 Penzel R, et al.EGFR mutation detection in NSCLC-assessment of diagnostic application and recommendations of the German Panel for Mutation Testing in NSCLC[J]. Virchows Arch,2011,458(1):95-98
158 Han HS. et al. EGFR mutation status in primary lung adenocarcinomas and corresponding metastatic lesions:discordance in pleural metastases [J]. Clin Lung Cancer,2011, 12(6):380-386
159 Gil-Bazo, IE. Castanon, JP Fusco, EGFR mutation testing in nonsmall cell lung cancer patients by using cytology specimens:when the tissue is no longer the issue[J]. Cancer Cytopathol,2011,119(5):354-355
160 Pfaff E, et al.TP53 mutation is frequently associated with CTNNB1 mutation or MYCN amplification and is compatible with long-term survival in medulloblastoma [J]. Clin Oncol,2010,28(35):5188-5196
161 Da SE, et al. TP53 mutation p. R337H in gastric cancer tissues of a 12 year old male child:evidence for chimerism involving a common mutant founder haplotype:case report[J].BMC Cancer,2011,11:449-450
162 Zenz T, et al.TP53 mutation profile in chronic lymphocytic leukemia:evidence for a disease specific profile from a comprehensive analysis of 268 mutations [J]. Leukemia,2010,24(12):2072-2079
163 Rossini A,et al.TP53 mutation profile of esophageal squamous cell carcinomas of patients from Southeastern Brazil[J].Mutat Res,2010,696 (1):10-15
164 Ruijs MW, et al.TP53 germLine mutation testing in 180 families suspected of Li-Fraumeni syndrome:mutation detection rate and relative frequency of cancers in different familial phenotypes [J].J Med Genet,2010,47(6):421-428
165 Amaravadi L,MJ Klemsz. DNA methylation and chromatin structure regulate PU.1 expression[J].DNA Cell Biol,1999,18 (12):875-884
166 Lu Q, et al. DNA methylation and chromatin structure regulate T cell per-forin gene expression[J]. Immunol,2003,170(10):5124-5132
167 Miller CA,SL Campbell,J D Sweatt, DNA methylation and his tone acetylation work in concert to regulate memory formation and synaptic plasticit [J]. Neurobiol Learn Mem,2008,89 (4):599-603
168 Tai KY,et al.DNA methylation and histone modification regulate silencing of epithelial cell adhesion molecule for tumor invasion and progress ion [J]. Oncogene, 2007,26(27):3989-3897
169 Lu TY,et al.DNA methylation and histone modification regulate silencing of OPG during tumor progression [J].Cell Biochem,2009,108 (1):315-325
170 Li W, et al.DNA methylation and histone modifications regulate shoot regeneration in Arabidopsis by modulating WUSCHEL expression and auxin signaling[J]. PLoS Genet,2011.7(8):e1002243
2 殷震,刘景华.动物病毒学.[M]第二版,北京：科学出版社,1997,836-894
3 邓普辉.绵羊肺腺瘤病一人细支气管-肺泡细胞癌的类似物.中国兽医科技,1988(2)：60-61
4 邓普辉.新疆绵羊肺腺瘤病调查[J].中国兽医杂志,1981(11)：4-5
5 幺宏强,其木格,马学恩.绵羊肺腺瘤病自然病例肺组织的超微结构观察[J].电子显微学报,2006：240
6 Grego E, et al. Colostrum and milk can transmit jaagsiekte retrovirus to lambs [J]. Vet Microbiol,2008,130 (3-4):247-257
7 Voigt K,et al. Eradication of ovine pulmonary adenocarcinoma by motherless rearing of lambs[J]. Vet Rec,2007,161 (4):129-132
8 Salvatori D,et al. Successful induction of ovine pulmonary adenocarcinoma in lamb of different ages and detection of viraemia during the preclinical period[J]. Gen Virol,2004.85(11):3319-3324
9 么宏强.绵羊肺腺瘤病毒人工感染羔羊淋巴细胞动态变化的研究[J].黑龙江畜牧兽医,2007(12)：69-71
10 Cousens C, et al. Evaluation of the efficacy of clinical diagnosis of ovine pulmonary adenocarcinoma[J]. Vet Rec,2008,162(3):88-90
11 Oda SS,SA Youssef. Immunohistochemical and histopathological findings of ovine pulmonary adenocarcinoma(Jaagsiekte) in Egyptian sheep[J].Trop Anim Health Prod, 2011,43(8):1611-1615
12 De Las HM, et al.A PCR technique for the detection of Jaagsiekte sheep retrovirus in the blood suitable for the screening of ovine pulmonary adenocarcinoma in field conditions [J].Res Vet Sci,2005,79 (3):259-264
13 Caporale M, et al.Expression of the jaagsiekte sheep retrovirus envelopegly sheep coprotein is sufficient to induce lung tumors in [J]. Virol,2006,80(16):8030-8037
14 Chitra E, et al. Functional interaction between env oncogene from Jagsiekte sheep retrovirus and tumor suppressor Sprouty2[J]. Retrovirology,2010(7):62
15 Palmarini M, et al. Jaagsiekte sheep retrovirus is necessary and sufficient toinduce a contagious lung cancer in sheep[J]. Virol,1999,73(8):6964-6972
16 Wootton SK,CL Halbert, AD Miller. Sheep retrovirus structural protein induceslung tumours[J]. Nature,2005,434 (7035):904-907
17 Miyazawa T.Receptors for animal retroviruses[J]. Uirusu,2009,59 (2):223-242
18 Vigdorovich V, RK Strong,AD Miller. Expression and characterization of a soluble, active form of the jaagsiekte sheep retrovirus receptor, Hya12[J]. Virol,2005, 79(1):79-86
19 Miller AD. Hyaluronidase 2 and its intriguing role as a cell-entry receptor for oncogenic sheep retroviruses [J].Semin Cancer Biol,2008,18 (4):296-301
20 Garcia-GotiM, et al.Sheep pulmonary adenomatosis:characterization of two patholo-gical forms associated with aagsiekte retrovirus [J]. Comp pthol,2000,122 (1):55-65
21 Maeda N, et al. Roles of the RAS-MEK-mi togen-act ivated protein kinaseand phosphate-idylinositol 3-kinase-AKT-mTOR pathways in Jaagsiekte sheep retrovirus-induced transformation of rodent fibroblast and epithelial cell lines[J]. Virol,2005, 79(7):4440-4450
22 Palmarini M, et al.A phosphat idylinositol 3-kinase docking site in the cytoplasmic tail of the Jaagsiekte sheep retrovirus transmembrane protein is essential for envelope induced transformation of NIH 3T3 cells[J]. Virol,2001,75 (22):11002-11009
23 De Las HM, et al. In-situ demonstration of mitogen-activated protein kinase Erk 1/2 signalling pathway in contagious respiratory tumours of sheep and goats[J]. Comp Pathol,2006,135(1):1-10
24 Liu SL, MI Lerman,AD Miller, Putative phosphatidylinositol 3-kinase (PI3K) binding motifs in ovine betaret rovirus env proteins are not essential for rodent fibroblast transformation and PI3K-AKT activation[J]. Virol,2003,77(14):7924-7935
25 Herring AJ,et al. Further evidence for a retrovirus as the aetiological agent of sheep pulmonary adenomatosis (jaagsiekte) [J]. Vet Microbiol,1983,8 (3):237-249
26 邓普辉,简子健,白惠敏.新疆绵羊肺腺瘤病回顾性研究[J].中国兽医杂志,1996(1)：3-5
27 林曦,郝先谱,赵振华.内蒙古动物肿瘤的调查[J].中国兽医病理学研究会肿瘤病理学专题讨论论文集,1985
28 Wander a JG. Pneumonia in sheep in Kenia. Part I Bacterial and parasitic pneumonia [J]. Bulletin of epizootic disease of Africa.1967.15:245-248
29 ustin RC.Ovine Jaagsiekte.Journal of the South African[J]. Veterinary Medical Association.1969,40:3-23
30 Sharp JM,Angus KW,Gray EW Scott. FMM. Rapid transmission of sheep pulmonary Adenomatosis(jaasiekte) in young lambs[J].Archives of Virology.1983,78:89-95
31 Niels Dungal.Experiments with Jaagsiekte[J]. Am J Pathol.1946,22 (4):737-759
32 Smith W,Mackay JMK. Morphological observations on a virus associated With sheep pulmonary adenomatosis(jaagsiekte) [J]. Journal of Comparative Pathology.1969,79: 421-425
33 Wandera JG. Sheep pulmonary adenomatosis (Jaagsiekte) [J].Advances in Veterinary cience and ComparativeMedicine.1971,15:251-283
34 Perk K, Hod I, Presentey B, et al.Lung carcinoma of sheep(jaagsiekte)[J].Journal of the National Cancer Institute.1971,47:197-205
35 Martin WB, Scott FMM,Sharp JM,Angus KW. Experimental production of sheep pulmonary adenomatosis (Jaagsiekte) [J].Nature.1976,264:183-184
36 Sharp JM, Herr ing AJ. Sheep pulmonary adenomatosis:demonstration of a protein which cross-reacts with the major core proteins of Mason-Pfizer monkey virus and mouse mammary tumour virus[J]. Gen Virol.1983,64:2323-2327
37 Myer MS, York DF,et al.The possible involvementof immunosuppression caused by a lentivirusin the aetiology of jaagsiekte and pasteurellosis in sheep [J]. Onderstepoort J Vet Res.1988,55:127-133
38 Verwoerd DW. Jaagsiekte (ovine pulmonary adenomatosis)virus. In:Dinta Z and Morein B(eds)[J]. Virus infections of ruminants.Elsevier,453-462
39 Afonso pv,et al. Centrosome and retroviruses:the dangerous liaisons [J]. Retrovirology,2007,4:27
40 Hecht SJ.et al.Distribution of endogenous type B and type D sheep retrovirus sequences in ungulates and other mammals[J]. Proc Natl Acad SciUSA,1996,93(8): 3297-3302
41 York F,Vigne R,Verwoerd DW, Querat G.Nucleotide sequence of the jaagsiekte retrovirus, an exogenous and endogenous type D and B retrovirus of sheep and goats [J]. Virol.1992,66 (8):4930-4939
42 Demartini JC, York D.Retrovirus-associated neoplasms of the respiratory system of sheep and goatsOvine pulmonary carcinoma and enzoot ic nasal tumor [J]. Vet Clin North Am Food Anim Pract.1997,13:55-70
43 中华人民共和国农业部公告动物疫病病种名录[M].《农业科技通讯》2000.10
44 Coil DA, et al. Jaagsiekte sheep retrovirus env protein stabilizes retrovirus vectors against inactivation by lung surfactant, centrifugation,and freeze-thaw cycling[J]. Virol,2001,75 (18):8864-8868
45 Cousens C, Thonur L, ImLach S. lung fluid produced by ovine pulmonary adenocarcinoma affected sheep and can survive for several weeks at ambient temperatures [J]. Res Vet Sci,2009,87(1):154-156
46 Summers C, et al. Jaagsiekte sheep retrovirus-specific immune responses induced by vaccination:a comparison of immunisation strategies [J]. Vaccine,2006,24(11): 1821-1829
47 Ortin A, et al.Lack of a specific immune response against a recombinant capsid protein of Jaagsiekte sheep retrovirus in sheep and goats naturally affected by enzootic nasal tumour or sheep pulmonary adenomatosis [J]. Vet Immunol Immunopathol, 1998,61(2-4):229-237
48 Hudachek SF,et al.Lung tumor development and spontaneous regression in lambs coinfected with Jaagsiekte sheep retrovirus and ovine lentivirus[J]. Vet Pathol, 2010,47(1):148-162
49 Cutlip RC,Young S, et al. Sheep pulmonary adenomatosis (jaagsiekte)in the United States [J]. American Journal of Veterinary Research.1982,43:2108-2113
50 Scott FMM, Sharp JM, Angus KW. Infection of specific-pathogen free lambs with a herpesvirus isolated from pulmonary adenomatosis [J].Archivo Virology.1984,80: 147-162
51 Holland MJ,et al.Jaagsiekte retrovirus is widely distributed both in T and B lymphocytes and in mononuclear phagocytes of sheep with naturally and experimentally acquired pulmonary adenomatosis [J].Virol,1999,73(5):4004-4008
52 Beytut EM,et al.Sozmen S Erginsoy. Immunohistochemical detection of pulmonary surfactant proteins and retroviral antigens in the lungs of sheep with pulmonary adenomatosis [J].Comp Pathol,2009,140(1):43-53
53 刘霄卉.绵羊肺腺瘤病毒囊膜基因表面蛋白的表达及其单克隆抗体的制备[J].中国预防兽医学报,2008(6)：469-472
54 Lewis FI, et al. Diagnostic accuracy of PCR for Jaagsiekte sheep retrovirus using field data from 125 Scottish sheep flocks[J].Vet J,2011,187 (1):104-108
55 Luo JR, et al. A specific PCR assay and a nested PCR assay for the screening of ovine pulmonary adenomatosis [J]. Bing Du Xue Bao,2009,25 (2):125-130
56 Voigt K, et al.PCR examination of bronchoalveolar lavage samples is a useful tool in pre-clinical diagnosis of ovine pulmonary adenocarcinoma (Jaagsiekte)[J].Res. Vet Sci,2007,83(3):419-427
57 Cousens C, et al.PCR-based detection and partial characterization of a retrovirus associated with contagious intranasal tumors of sheep and goats[J]. Virol,1996, 70(11):7580-7583
58 Gonzalez L, et al. Jaagsiekte sheep retrovirus can be detected in the peripheral blood during the pre-clinical period of sheep pulmonary adenomatosis [J]. Gen Virol, 2001,82(6):1355-1358
59 Sanna MP, et al. Association of jaagsiekte sheep retrovirus with pulmonary carcinoma in Sardinian moufflon (Ovis musimon) [J]. Comp Pathol,2001,125(2-3):145-152
60 Morozov VA,et al.Detection and characterization of betaretroviral sequences, related to sheep Jaagsiekte virus, in Africans from Nigeria and Cameroon[J]. Virology,2004,327 (2):162-168
61 Fox KA,et al.Paranasal sinus masses of Rocky Mountain bighorn sheep (Ovis canadensis Canadensis)[J]. Vet Pathol,2011,48(3):706-712
62 Maeda N, et al. Surveillance of Jaagsiekte sheep retrovirus in sheep in Hokkaido, the northern island of Japan[J].Vet Med Sci,2011,73(11):1493-1495
63 Suau F, et al. TelomeRASe activation in a model of lung adenocarcinoma. Eur Respir [J]. 2006,27(6):1175-1182
64 Palmarini M, H Fan, JM Sharp. Sheep pulmonary adenomatosis:a unique model of retrovirus associated lung cancer [J].Trends Microbiol,1997,5(12):478-483
65 Palmarini M, H Fan. Retrovirus-induced ovine pulmonary adenocarcinoma,an animal model for lung cancer[J].Natl Cancer Inst,2001,93 (21):1603-1614
66 Occa S, et al. Presence of Jaagsiekte sheep retrovirus in tissue sections from human bronchioloalveolar carcinoma depends on patients'geographical origin[J].Hum Pathol,2008,39 (2):303-304
67 Mornex JF,et al.Pathology of human bronchioloalveolar carcinoma and its relationship to the ovine disease[J].Curr Top Microbiol Immunol,2003,275:225-248
68 Hurnann-Ziehank E, et al. Ovine pulmonary adenocarcinoma as an animal model of progressive lung cancer and the impact of nutritional selenium supply [J]. Trace Elem Med Biol,2011,25 (11):30-34
69 Murgia C, et al.Lung adenocarcinoma originates from retrovirus infection of proferating type 3 pneumocytes during pulmonary post-natal development or tissue repair[J].PLoS Pathog,2011,7(3):100-114
70 Hiatt KM, WE Highsmith. Lack of DNA evidence for jaagsiekte sheep retrovirus in human bronchioloalveolar carcinoma[J]. Hum Pathol,2002,33 (6):680
71 Yousem SA,et al.Absence of jaagsiekte sheep retrovirus DNA and RNA in bronchioloalveolar and conventional human pulmonary adenocarcinoma by PCR and RT-PCR analysis[J]. Hum Pathol,2001,32 (10):1039-1042
72 Hopwood P, et al. Absence of markers of betaretrovirus infection in human pulmonary adenocarcinoma [J]. Hum Pathol,2010,41 (11):1631-1640
73 DeMartini JC,et al.Jaagsiekte sheep retrovirus proviral clone JSRV(JS7), derived from the JS7 lung tumor cell line, induces ovine pulmonary carcinoma and is integrated into the surfactant protein A gene[J]. Virol,2001,75(9):4239-4246
74 Ni tta T, et al. Identification and mutational analysis of a Rej response element in jaagsiekte sheep retrovorus RNA[J]. Virol,2009,83(23):12499-12511
75 Chitra E, Yu SL, Hsiao KN,Shao HY. Generation and characterization of JSRV envelope transgenic mice in FVB background[J]. Virology.2009,393(1):120-126
76 Ortin A, et al. Characterization of enzootic nasal tumour virus of goats:complete sequence and tissue distribution[J].Gen Virol,2003.84:2245-2252
77 Cousens C, et al. Complete sequence of enzootic nasal tumor virus,a retrovirus associated with transmissible intranasal tumors of sheep[J]. Virol,1999,73(5): 3986-3993
78 Kaufmann C, et al. Enzootic nasal adenocarcinoma in a dairy sheep flock [J]. Schweiz Arch TierheilkD,2008,150(6):297-302
79 De Las, et al. Enzootic nasal adenocarcinoma of sheep and goats [J]. Curr Top Microbiol Immunol,2003,275,201-223
80 Wootton SK.CLHalbert, AD Miller. envelope proteins of Jaagsiekte sheep retrovirus and enzootic nasal tumor virus induce similar bronchioalveolar tumors in lungs of mice[J]. Virol,2006,80(18):9322-9325
81 Dunlap KA,M Palmarini, TE Spencer, Ovine endogenous betaretroviruses (enJSRV) and placental morphogenesis. Placenta,2006,27:135-140
82 Carlson J, et al. Chromosomal distribution of endogenous Jaagsiekte sheep retrovirus proviral sequences in the sheep genome [J]. Virol,2003.77 (17):9662-9668
83 Hecht SJ,JO.Carlson,JC DeMartin. Analysis of a type D retroviral capsid gene expressed in ovine pulmonary carcinoma and present in both affected and unaffected sheep genomes[J]. Virology,1994,202(1):480-484
84 Mortuza GB,et al. Structure of the capsid amino-terminal domain from the betaretrovirus, Jaagsiekte sheep retrovirus[J].J Mol Biol,2009,386 (4):1179-1192
85 Johnson C, et al. Enhanced proliferation of primary rat type Ⅱ pneumocytes by Jaagsiekte sheep retrovirus envelope protein[J].Virology,2011,412 (2):349-355
86 Golovkina TV,Jaffe AB,Ross SR. Coexpression of exogenous and endogenous mouse mammary tumour virus RNA in vivo results in viral recombination and broadens the virus host range [J].J Virol,1994,68:5019-5026
87 Golovkina TV, Chervonsky A.Dudley JP, Ross SR. Transgenic mouse mammary tumor virus superantigen expression prevents viral infection[J]. Cell,1992,69:637-645
88 Berkowitz R, Fisher J.Goff SP. RNA packaging [J]. Curr Top Microbiol Immunol 1996,214: 177-218
89 Rosati S,et al.An accessory open reading frame (orf-x) of jaagsiekte sheep retrovirus is conserved between different virus isolates[J]. Virus Res,2000,66(1): 109-116
90 McGee-Estrada K, H Fan, Comparison of LTR enhancer elements in sheep beta retroviruses:insights into the basis for tissue-specific expression[J]. Virus Genes,2007,35(2):303-312
91 McGee-Estrada K,et al.A Moloney murine leukemia virus driven by the Jaagsiekte sheep retrovirus enhancers shows enhanced specificity for infectivity in lung epithelial cells[J]. Virus Genes,2005,31 (3):257-263
92 McGee-Estrada KM, Palmarini H Fan. HNF-3beta is a critical factor for the expression of the Jaagsiekte sheep retrovirus long terminal repeat in type II pneumocytes but not in Clara cells[J]. Virology 2002,292(1):87-97
93 Sinn PL, et al. Inclusion of Jaagsiekte sheep retrovirus proviral elements markedly increases lentivirus vector pseudotyping efficiency [J]. Mol Ther,2005,11 (3):460-469
94 McGee-Estrada K, H Fan. In vivo and in vitro analysis of factor binding sites in Jaagsiekte sheep retrovirus long terminal repeat enhancer sequences:roles of HNF-3, NF-I, and C/EBP for activity in lung epithelial cells[J]. Virol,2006,80(1):332-341
95 Dakessian RM, H Fan. Specific in vivo expression in type II pneumocytes of the Jaagsiekte sheep retrovirus long terminal repeat in transgenic mice[J]. Virology, 2008,372(2):398-408
96 Palmarini M,et al.The long terminal repeat of Jaagsiekte sheep retrovirus is preferentially active in differentiated epithelial cells of the lungs [J]. Virol, 2000,74(13):5776-5787
97 Leroux C,et al.Jaagsiekte Sheep Retrovirus (JSRV):from virus to lung cancer in sheep [J]. Vet Res,2007,38 (2):211-228
98 Philbey AW,et al. Multiclonal pattern of Jaagsiekte sheep retrovirus integration sites in ovine pulmonary adenocarcinoma [J]. Virus Res,2006,117 (2):254-263
99 Cousens C,et al. Analysis of integration sites of Jaagsiekte sheep retrovirus in ovine pulmonary adenocarcinoma[J]. J Virol,2004.78 (16):8506-8512
100 Rai SK,Miller AD. Retrovirus vector bearing jaagsiekte sheep retrovirus env transducer human cells by using a new receptor localized to chromosome 3p21.3 [J]. Virol 2000,74:4698-4704
101 Vigdorovich V, AD Miller. Ability of hyaluronidase2 to degrade extracellular hyaluronan is not required for its function as a receptor for jaagsiekte sheep retrovirus [J].Virol,2007,81(7):3124-4129
102 Duh FM, et al. Amino acid residues that are important for Hya12 function as a receptor for jaagsiekte sheep retrovirus [J].Retrovirology,2005,2:59
103 Miller AD.Identification of Hya12 as the cell-surface receptor for jaagsiekte sheep retrovirus and ovine nasal adenocarcinoma virus [J]. CurrTop Microbiol Immunol, 2003,275:179-199
104 Van Hoeven NS, AD Miller. Improved enzootic nasal tumor virus pseudotype packaging cell lines reveal virus entry requirements in addition to the primary receptor Hya12[J]. Virol,2005,79(1):87-94
105 Rai SK, JC DeMartini.AD Miller. Retrovirus vectors bearing jaagsiekte sheep retrovirus env transduce human cells by using a new receptor localized to chromosome 3p21.3[J].Virol,2000,74(10):4698-4704
106 Liu SL, et al.Role of virus receptor Hya12 in oncogenic transformation of rodent fibroblasts by sheep betaretrovirus env proteins[J], Virol,2003,77(5):2850-2858
107 Sharath K Rai,Fuh-Mei Duh, Vladimir Vigdorovich,et al. Candidate tumor suppressor HYAL2 is a glycosylphosphatidylinositol (GPI)-anchored cell-surface receptor for jaagsiekte sheep retrovirus, the envelope protein of which mediates oncogenic transformation[J].2001,98 (8):4443-4448
108 Palmarini M.M Mura.TE Spencer. Endogenous betaretroviruses of sheep:teaching new lessons in retroviral interference and adaptation[J].J Gen Virol,2004,85 (Ptl): 1-13
109 Sanna E, et al. Endogenous jaagsiekte sheep retrovirus RNA is expressed by different cell types in ovine foetus and placenta[J]. Eur J Histochem,2002,46(3):273-280
110 Dunlap KA,et al.Endogenous retroviruses regulate periimplantation placental growth and differentiation[J]. Proc Natl Acad Sci USA,2006.103(39):14390-14395
111 DeMartini JC. Endogenous retroviruses related to jaagsiekte sheep retrovirus [J]. Curr Top Microbiol Immunol,2003,275:117-137
112 Spencer TE,M Palmarini. Endogenous retroviruses of sheep:a model system for understanding physiological adaptation to an evolving ruminant genome [J]. J Reprod Dev,2012.58(1):33-37
113 Palmarini M,et al.Molecular cloning and functional analysis of three type D endogenous retroviruses of sheep reveal a different cell tropism from that of the highly related exogenous jaagsiekte sheep retrovirus [J]. Virol,2000,74 (17):8065-8076
114 Black SG.et al. Endogenous retroviruses in trophoblast differentiation and placental development [J]. Am J Reprod Immunol 2010,64(4):255-264
115 Arnaud F,et al. Coevolution of endogenous betaretroviruses of sheep and their host[J].Cell Mol Life Sci,2008,65(21):3422-3432
116 Arnaud F,et al.A paradigm for virus-host coevolution:sequential counter-adaptations between endogenous and exogenous retroviruses [J].PLoS Pathog,2007, 3(11):e170
117 Frederick Arnaud.Marco Caporale,Mariana Varela. A Paradigm for Virus Host Coevolution:Sequential Counter-Adaptations between Endogenous and Exogenous Retroviruses [J]. PLoS Pathog,2007,3(11):e170
118 Kathrin A, Dunlap, Massimo Palmarini. Endogenous retroviruses regulate periimp-lantation placental growth and differentiation[J]. Proc Natl Acad Sci USA.2006, 103(39):165-176
119 Mura M, et al. Late viral interference induced by transdominant Gag of an endogenous retrovirus[J].Proc Natl Acad Sci USA,2004.101 (30):11117-11122
120 Arnaud F, PR Murcia,M Palmarini,Mechanisms of late restriction induced by an endogenous retrovirus [J]. Virol,2007,81 (20):11441-11451
121 Spencer TE, Mariana Varela, et al.Receptor usage and fetal expression of ovine endogenous betaret roviruses:implications for coevolution of endogenous and exogenous retroviruses[J]. Virol,2003,77 (1):749-753
122 Armezzani A, et al.The signal peptide of a recently integrated endogenous sheep betaretrovirus envelope plays a major role in eluding gag-mediated late restriction[J].Virol,2011,85 (14):7118-7128
123 Murcia PR,F Arnaud, M Palmarini, The transdominant endogenous retrovirus enJS56Al associates with and blocks intracellular trafficking of Jaagsiekte sheep retrovirus Gag[J]. Virol,2007,81 (4):1762-1772
124 Massimo Palmarini, Naoyoshi Maeda, Claudio Murgia, et al.A Phosphatidy linositol 3-Kinase Docking Site in the Cytoplasmic Tail of the Jaagsiekte Sheep Retrovirus Transmembrane Protein Is Essential for envelope-Induced Transformation of NIH 3T3 Cells[J]. Virol.2001,75 (22):11002-11009
125 Marceline C, Yi-Min Zheng, Lorraine M. Albritton, Fusogenicity of Jaagsiekte Sheep Retrovirus envelope Protein Is Dependent on Low pH and Is Enhanced by Cytoplasmic Tail Truncations [J].Virol.2008,82(5):2543-2554
126 Sharath K, Rai,Fuh-Mei Duh,Vladimir Vigdorovich,et al. Candidate tumor suppressor HYAL2 is a glycosylphosphatidylinositol (GPI)-anchored cell-surface receptor for jaagsiekte sheep retrovirus, the envelope protein of which mediates oncogenic transformation [J].Proc Natl Acad Sci USA,2001,98 (8):4443-4448
127 Miller AD,NS Van Hoeven,SL Liu. Transformation and scattering activities of the receptor tyrosine kinase RON/Stk in rodent fibroblasts and lack of regulation by the jaagsiekte sheep retrovirus receptor [J], BMC Cancer,2004,4:64
128 Danilkovitch-Miagkova A, et al. Hyaluronidase 2 negatively regulates RON receptor tyros ine kinase and mediates transformat ion of epithelial cells by jaagsiekte sheep retrovirus[J].Proc Natl Acad Sci USA,2003,100(8):4580-4585
129 Hull, S, H Fan.Mutational analysis of the cytoplasmic tail of jaagsiekte sheep retrovirus envelope protein[J]. Virol,2006,80(16):8069-8080
130 Massimo Palmarini,J Michael Sharp, Christine Lee. In Vitro Infection of Ovine Cell Lines by Jaagsiekte Sheep Retrovirus[J]. Virol,1999,73 (12):10070-10078
131 Hofacre A, H Fan. Multiple domains of the Jaagsiekte sheep retrovirus envelope protein are required for transformation of rodent fibroblasts[J]. Virol,2004,78: 10479-10489
132 Hallwirth C, et al. Variable regions 1 and 2 (VR1 and VR2) in JSRV gag are not responsible for the endogenous JSRV particle release defect[J]. Virus Genes,2005, 30(1):59-68
133 Naoyoshi Maeda,Wuxia Fu, Aurora Ortin. Roles of the RAS-MEK-Mitogen-Activated Protein Kinase and Phosphatidylinositol 3-Kinase-AKT-mTOR Pathways in Jaagsiekte Sheep Retrovirus-Induced Transformation of Rodent Fibroblast and Epithelial Cell Lines[J]. Virol,2005,79 (7):4440-4450
134 Mariana Varela, Matthew Golder, et al.A Large Animal Model to Evaluate the Effects of Hsp90 Inhibitors for the Treatment of Lung Adenocarcinoma [J]. Virology,2008, 371(1):206-215
135 Johnson C,K Sanders,H Fan. Jaagsiekte sheep retrovirus transformation in Madin Darby canine kidney epithelial cell three-dimensional culture[J].J Virol,2010, 84(10):5379-5390
136 Dani lkovi tch-Miagkova, AFM Duh, I Kuzmin, D Angeloni,SL Liu, AD Miller, MI Lerman. Hyaluronidase2 negatively regulates RON receptor tyrosine kinase and mediates transformation of epithelial cells by jaagsiekte sheep retrovirus[J]. Natl Acad Sci.USA 2003,100:4580-4585
137 Vaughan A E, et al.Lung cancer in mice induced by the jaagsiekte sheep retrovirus envelope protein is not maintained by rare cancer stem cells, but tumorigenicity does correlate with Wnt pathway activation[J]. Mol Cancer Res,2012,10 (1):86-95
138 Maeda N,et al.Transformation of mouse fibroblasts by Jaagsiekte sheep retrovirus envelope does not require phosphatidylinositol 3-kinase[J]. Virol,2003,77 (18): 9951-9959
139 Maeda N,et al Direct transformation of rodent fibroblasts by jaagsiekte sheep retrovirus DNA[J].Proc Natl Acad Sci USA,2001,98 (8):4449-4454
140 Shan-Lu Liu A, Dusty Miller. Transformation of Madin-Darby Canine Kidney Epithelial Cells by Sheep Retrovirus envelope Proteins [J].J Virol,2005,79 (2):927-933
141 Chetaille MC, S Olschwang, KRAS Mutation Spectrum Notably Diverges between Non-small Cell Lung and Colorectal Carcinomas[J].Thorac Oncol,2012.7 (4):773-774
142刘丽琴,张海燕,王捷KRAS基因突变检测与结直肠癌分子靶向治疗的研究进展[J].现代肿瘤医学,2011(4)：802-804
143 Jing Gao, Tingting Wang, Jing wei Yu, Yan-yan Li. Wild-Type KRAS and BRAF Could Predict Response to Cetuximab in Chinese Colorectal Cancer Patients[J]. Chinese Journal of Cancer Research,2011 (4):271-275
144 Ellis PM. KRAS Mutation Spectrum Notably Diverges between Non-small Cell Lung and Colorectal Carcinomas [J].Thorac Oncol,2012.7 (4):774
145隆艳艳,戴广海.胰腺癌中EGFR、KRAS及PI3K/AKT通路对EGFR-TKIs疗效预测的研究[J].军医进修学院学报,2011(11)：1185-1187
146 Buttitta F,et al.PIK3CA mutation and histological type in breast carcinoma:high frequency of mutations in lobular carcinoma [J]. Pathol,2006,208 (3):350-355
147 Kalinsky K,et al.PIK3CA mutation associates with improved outcome in breast cancer[J].Clin Cancer Res,2009,15(16):5049-5059
148 Cizkova M, et al. PIK3CA mutation impact on survival in breast cancer patients and in ERalpha, PR and ERBB2-based subgroups[J]. Breast Cancer Res,2012,14 (1):R28
149 Nosho K, et al. PIK3CA mutation in colorectal cancer:relationship with genetic and epigenetic alterations [J]Neoplasia,2008,10(6):534-541
150 Yamamoto S. PIK3CA mutation is an early event in the development of endometrios is associated ovarian clear cell adenocarcinoma [J]. Pathol,2011,225 (2):189-194
151 Kozaki K, et al.PIK3CA mutation is an oncogenic aberration at advanced stages of oral squamous cell carcinoma [J].Cancer Sci,2006,97 (12):1351-1358
152 Ogino S, et al. PIK3CA mutation is associated with poor prognosis among patients with curatively resected colon cancer[J]. Clin Oncol,2009.27 (9):1477-1484
153 Kato S,et al.PIK3CA mutation is predictive of poor survival in patients with colorectal cancer [J].Int J Cancer,2007,121(8):1771-1778
154 Thunnisse E, et al. EGFR-mutation in non-small cell lung carcinoma. TreaTment with tyrosine kinase inhibitors possible[J]. Ned Tijdschr GeneeskD,2011,155:A2554
155 Li F, et al.Role of EGFR mutation status in patients with stage Ⅲ non-squamous non-small cell lung cancer treated with chemoradiotherapy [J].Zhongguo Fei Ai Za Zhi,2011,14(9):715-718
156 Saitoh M,et al.An early event of EGFR mutation in pleomorphic carcinoma of the lung[J].Int J Clin Oncol,2011,16 (6):770-773
157 Penzel R, et al.EGFR mutation detection in NSCLC-assessment of diagnostic application and recommendations of the German Panel for Mutation Testing in NSCLC[J]. Virchows Arch,2011,458(1):95-98
158 Han HS. et al. EGFR mutation status in primary lung adenocarcinomas and corresponding metastatic lesions:discordance in pleural metastases [J]. Clin Lung Cancer,2011, 12(6):380-386
159 Gil-Bazo, IE. Castanon, JP Fusco, EGFR mutation testing in nonsmall cell lung cancer patients by using cytology specimens:when the tissue is no longer the issue[J]. Cancer Cytopathol,2011,119(5):354-355
160 Pfaff E, et al.TP53 mutation is frequently associated with CTNNB1 mutation or MYCN amplification and is compatible with long-term survival in medulloblastoma [J]. Clin Oncol,2010,28(35):5188-5196
161 Da SE, et al. TP53 mutation p. R337H in gastric cancer tissues of a 12 year old male child:evidence for chimerism involving a common mutant founder haplotype:case report[J].BMC Cancer,2011,11:449-450
162 Zenz T, et al.TP53 mutation profile in chronic lymphocytic leukemia:evidence for a disease specific profile from a comprehensive analysis of 268 mutations [J]. Leukemia,2010,24(12):2072-2079
163 Rossini A,et al.TP53 mutation profile of esophageal squamous cell carcinomas of patients from Southeastern Brazil[J].Mutat Res,2010,696 (1):10-15
164 Ruijs MW, et al.TP53 germLine mutation testing in 180 families suspected of Li-Fraumeni syndrome:mutation detection rate and relative frequency of cancers in different familial phenotypes [J].J Med Genet,2010,47(6):421-428
165 Amaravadi L,MJ Klemsz. DNA methylation and chromatin structure regulate PU.1 expression[J].DNA Cell Biol,1999,18 (12):875-884
166 Lu Q, et al. DNA methylation and chromatin structure regulate T cell per-forin gene expression[J]. Immunol,2003,170(10):5124-5132
167 Miller CA,SL Campbell,J D Sweatt, DNA methylation and his tone acetylation work in concert to regulate memory formation and synaptic plasticit [J]. Neurobiol Learn Mem,2008,89 (4):599-603
168 Tai KY,et al.DNA methylation and histone modification regulate silencing of epithelial cell adhesion molecule for tumor invasion and progress ion [J]. Oncogene, 2007,26(27):3989-3897
169 Lu TY,et al.DNA methylation and histone modification regulate silencing of OPG during tumor progression [J].Cell Biochem,2009,108 (1):315-325
170 Li W, et al.DNA methylation and histone modifications regulate shoot regeneration in Arabidopsis by modulating WUSCHEL expression and auxin signaling[J]. PLoS Genet,2011.7(8):e1002243