鼻咽癌眼部症状及联合转染siRNA对鼻咽癌细胞基因抑制的研究
|
摘要
目的:1、临床研究的目的在于探讨鼻咽癌眼部症状的临床表现和影像学及病理学诊断及相互对应的关系,意义在于鼻咽癌眼部的症状虽不多见,但也占一定比例,在临床工作中容易被忽视造成误诊和漏诊,在鼻咽癌高发区应对眼部症状进行鼻咽癌的排查,为鼻咽癌的早期诊断提供依据。2、实验研究在于探讨单独和联合转染siRNA(hTERTsiRNA,PCNAsiRNA)技术对人鼻咽癌CNE2细胞相关基因表达以及细胞增殖的抑制作用,同时探讨此技术对CNE2细胞的细胞周期和凋亡的影响,以及对相关蛋白表达的抑制作用,旨在为鼻咽癌的治疗提供新思路和方法。
方法:1、临床研究部分:选择2003-2008年6年间562名鼻咽癌患者,对其中82名有眼部症状患者的临床表现,病理诊断和CT表现进行回顾性分析。2、实验研究部分:1)hTERT和PCNAsiRNA的序列设计、合成和筛选。2)分别单独转染hTERTsiRNA、PCNAsiRNA,以及联合转染hTERTsiRNA和PCNAsiRNA:通过MMT法观察细胞增殖情况,流式细胞仪检测细胞周期的变化,实时定量PCR检测细胞相关基因的沉默效应,倒置相差显微镜和免疫组织化学检测细胞生长及相关基因的蛋白表达。
结果:1、临床研究显示眼部症状的NPC患者占全部NPC患者的14.6%(82/562)。男性64人(70眼),女性18人(21眼)。患者平均年龄为50.5岁(26-78岁)。累及双眼者9人。来自广东省65例,湖南省6例,江西省4例,广西省4例,海南省2例,湖北省1例。视朦32例(37眼),复视30例(31眼),眼球运动受限25例(26眼),视力下降12例(14眼),眼睑下垂8例,失明6例,眼眶肿胀3例,眼球突出3例,眼痛1例。累及左眼36例,右眼37例,双眼9例。颅底骨质破坏及颅内侵犯40例(包括卵圆孔、棘孔、岩尖、斜坡、蝶窦基底、蝶骨大翼、翼内外板、枕骨大孔、颈静脉孔、海绵窦);侵犯眼眶6例;眶内转移2例(左眼);累及蝶窦6例,上颌窦3例,筛窦3例。咽旁间隙68例,茎突内侧软组织49例。(同一病例可有多种表现,数据有重叠)
2、实验研究显示(1)siRNA单独和联合转染都能在不同程度有效抑制hTERT、PCNA及RB1相关基因的表达及细胞增殖:(2)转染48h内未观察到细胞凋亡,但细胞静止于G0/G1期,肿瘤细胞的增殖受到抑制;(3)单独和联合转染后CNE2细胞中hTERT、PCNA及RB1蛋白表达明显受到抑制。(4)联合转染在对CNE2细胞的hTERT、PCNA及RB1基因表达和细胞增殖的抑制作用以及对CNE2细胞的周期影响以及相关蛋白表达的抑制作用优于单独转染,差异有统计学意义。
结论:1、鼻咽癌眼部症状具有多样化和复杂性,在临床工作中应引起足够重视。2、siRNA(hTERT、PCNA)单独和联合转染对CNE2细胞的相关基因和蛋白及细胞增殖、细胞周期都有干扰效应,联合转染的siRNA干扰作用优于单独转染。为鼻咽癌的基因治疗提供新的思路和方法。
Objective:1.The purpose of clinical study is to explore the eye complication of nasopharyngeral carcinoma(NPC),to analyses the clinical manifestation,CT characteristics and pathological diagnosis of eye complications of NPC,to provide the base for early diagnosis of NPC.2.The purpose of experiment study is to explore the effect of separate and combining hTERTsiRNA,PCNAsiRNA on NPC CNE2 cell;to explore whether targeting siRNA(hTERT, PCNA separate and combining) could inhibit the expressing of related gene and related protein of NPC CNE2 cell,and tumor cells growth,meantime to explore the effect of targeting siRNA (hTERT,PCNA separate and combining ) to cell cycle and apoptosis of NPC CNE2 cell,and to discuss the future application on siRNA in the gene therapy of NPC.
Methods:1.The part of clinical study:to retrospective study of 82 cases eye complications in 562 cases NPC,to study their clinical manifestation,CT characteristics and pathological diagnosis.2. The part of experiment study:l) to designing,synthesis and screen effective siRNA ofhTERT and PCNA.2) Effective hTERTsiRNA,PCNAsiRNA separate and combining was transfected into NPC CNE2 cell line by using Lip2000TM siRNA transfection reagent.The relate mRNA and protein were detected by Real-time polymerase chain reaction(Real-time PCR) and immunohistochemistry respectively.MTT assay was used to determine the inhibitory effect of siRNA on tumor cells growth and inverted phase contrast microscope and flow cytometry were used to observe the cell growth,cell cycle,and the apoptosis induced by siRNA.
Results:1.The clinical studies showed that eye complication cases were 82cases of 562 NPC cases(14.6%),there were male 64 cases(70eyes) and female 18cases(21 eyes),The mean age at diagnosis was 50.5 years.The bilateral eyes were involved in 9cases.65cases came from Guangdong,6cases from Hunan,4cases from Jiangxi,4cases from Guangxi,2cases from Hainan, 1 cases from Hubei.32cases(37eyes) with blurred vision,30cases(31 eyes) with diplopia, 25cases(26eyes) with ocular movement disorder,12cases(14eyes)with decrease of visual acuity, 8cases with ptosis,6cases with acroisa,3cases with fossa orbitalis swelling,3cases with exophthalmos,1case with ophthalmalgia,36 cases in left and 37 cases in right eyes,9 cases in bilateral eyes.CT appearances:40cases suffered from skull base distracted,6cases with orbit involved,2cases(in left eyes) with orbit metastasis,6cases with sphenoidal sinus involved, 3cases maxillary sinus and 3cases ethmoidal sinus,68case with parapharyngeal space involved, 49cases with soft issue in wall of styloid process involved(there were many kind of shows in th same case,so the data were repeated in these cases).
2.The experimental studies showed that 1) Separate and combining siRNA(hTERT,PCNA) transfected could inhibit the expression of hTERT,PCNA and RB1 gene,and cell growth effectually.2) Apoptosis were not observed in transfected CNE2 cells after 48h,but cell cycle stasis in G0/G1 stage.3) Separate and combining siRNA transfected could inhibit the expressio of hTERT,PCNA and RB1 protein obviously.4) Combining siRNA transfected was better than separate siRNA transfected in inhibiting the expression of related gene,protein and cell cycle growth,the difference was significant.
Conclusion:1.As there were multiplicity and complexity in eye complication of NPC,so clinician should think highly of these cases.2.Separate and combining siRNA transfected (hTERT,PCNA) could inhibit the expression of related gene and protein of CNE2 cell,and interfere cell growth and cell cycle effectually,combining siRNA transfected was better than separate siRNA transfected in interfering effects to NPC CNE2 cell.This study provides new thoughts and methods for NPC gene therapy.
方法:1、临床研究部分:选择2003-2008年6年间562名鼻咽癌患者,对其中82名有眼部症状患者的临床表现,病理诊断和CT表现进行回顾性分析。2、实验研究部分:1)hTERT和PCNAsiRNA的序列设计、合成和筛选。2)分别单独转染hTERTsiRNA、PCNAsiRNA,以及联合转染hTERTsiRNA和PCNAsiRNA:通过MMT法观察细胞增殖情况,流式细胞仪检测细胞周期的变化,实时定量PCR检测细胞相关基因的沉默效应,倒置相差显微镜和免疫组织化学检测细胞生长及相关基因的蛋白表达。
结果:1、临床研究显示眼部症状的NPC患者占全部NPC患者的14.6%(82/562)。男性64人(70眼),女性18人(21眼)。患者平均年龄为50.5岁(26-78岁)。累及双眼者9人。来自广东省65例,湖南省6例,江西省4例,广西省4例,海南省2例,湖北省1例。视朦32例(37眼),复视30例(31眼),眼球运动受限25例(26眼),视力下降12例(14眼),眼睑下垂8例,失明6例,眼眶肿胀3例,眼球突出3例,眼痛1例。累及左眼36例,右眼37例,双眼9例。颅底骨质破坏及颅内侵犯40例(包括卵圆孔、棘孔、岩尖、斜坡、蝶窦基底、蝶骨大翼、翼内外板、枕骨大孔、颈静脉孔、海绵窦);侵犯眼眶6例;眶内转移2例(左眼);累及蝶窦6例,上颌窦3例,筛窦3例。咽旁间隙68例,茎突内侧软组织49例。(同一病例可有多种表现,数据有重叠)
2、实验研究显示(1)siRNA单独和联合转染都能在不同程度有效抑制hTERT、PCNA及RB1相关基因的表达及细胞增殖:(2)转染48h内未观察到细胞凋亡,但细胞静止于G0/G1期,肿瘤细胞的增殖受到抑制;(3)单独和联合转染后CNE2细胞中hTERT、PCNA及RB1蛋白表达明显受到抑制。(4)联合转染在对CNE2细胞的hTERT、PCNA及RB1基因表达和细胞增殖的抑制作用以及对CNE2细胞的周期影响以及相关蛋白表达的抑制作用优于单独转染,差异有统计学意义。
结论:1、鼻咽癌眼部症状具有多样化和复杂性,在临床工作中应引起足够重视。2、siRNA(hTERT、PCNA)单独和联合转染对CNE2细胞的相关基因和蛋白及细胞增殖、细胞周期都有干扰效应,联合转染的siRNA干扰作用优于单独转染。为鼻咽癌的基因治疗提供新的思路和方法。
Objective:1.The purpose of clinical study is to explore the eye complication of nasopharyngeral carcinoma(NPC),to analyses the clinical manifestation,CT characteristics and pathological diagnosis of eye complications of NPC,to provide the base for early diagnosis of NPC.2.The purpose of experiment study is to explore the effect of separate and combining hTERTsiRNA,PCNAsiRNA on NPC CNE2 cell;to explore whether targeting siRNA(hTERT, PCNA separate and combining) could inhibit the expressing of related gene and related protein of NPC CNE2 cell,and tumor cells growth,meantime to explore the effect of targeting siRNA (hTERT,PCNA separate and combining ) to cell cycle and apoptosis of NPC CNE2 cell,and to discuss the future application on siRNA in the gene therapy of NPC.
Methods:1.The part of clinical study:to retrospective study of 82 cases eye complications in 562 cases NPC,to study their clinical manifestation,CT characteristics and pathological diagnosis.2. The part of experiment study:l) to designing,synthesis and screen effective siRNA ofhTERT and PCNA.2) Effective hTERTsiRNA,PCNAsiRNA separate and combining was transfected into NPC CNE2 cell line by using Lip2000TM siRNA transfection reagent.The relate mRNA and protein were detected by Real-time polymerase chain reaction(Real-time PCR) and immunohistochemistry respectively.MTT assay was used to determine the inhibitory effect of siRNA on tumor cells growth and inverted phase contrast microscope and flow cytometry were used to observe the cell growth,cell cycle,and the apoptosis induced by siRNA.
Results:1.The clinical studies showed that eye complication cases were 82cases of 562 NPC cases(14.6%),there were male 64 cases(70eyes) and female 18cases(21 eyes),The mean age at diagnosis was 50.5 years.The bilateral eyes were involved in 9cases.65cases came from Guangdong,6cases from Hunan,4cases from Jiangxi,4cases from Guangxi,2cases from Hainan, 1 cases from Hubei.32cases(37eyes) with blurred vision,30cases(31 eyes) with diplopia, 25cases(26eyes) with ocular movement disorder,12cases(14eyes)with decrease of visual acuity, 8cases with ptosis,6cases with acroisa,3cases with fossa orbitalis swelling,3cases with exophthalmos,1case with ophthalmalgia,36 cases in left and 37 cases in right eyes,9 cases in bilateral eyes.CT appearances:40cases suffered from skull base distracted,6cases with orbit involved,2cases(in left eyes) with orbit metastasis,6cases with sphenoidal sinus involved, 3cases maxillary sinus and 3cases ethmoidal sinus,68case with parapharyngeal space involved, 49cases with soft issue in wall of styloid process involved(there were many kind of shows in th same case,so the data were repeated in these cases).
2.The experimental studies showed that 1) Separate and combining siRNA(hTERT,PCNA) transfected could inhibit the expression of hTERT,PCNA and RB1 gene,and cell growth effectually.2) Apoptosis were not observed in transfected CNE2 cells after 48h,but cell cycle stasis in G0/G1 stage.3) Separate and combining siRNA transfected could inhibit the expressio of hTERT,PCNA and RB1 protein obviously.4) Combining siRNA transfected was better than separate siRNA transfected in inhibiting the expression of related gene,protein and cell cycle growth,the difference was significant.
Conclusion:1.As there were multiplicity and complexity in eye complication of NPC,so clinician should think highly of these cases.2.Separate and combining siRNA transfected (hTERT,PCNA) could inhibit the expression of related gene and protein of CNE2 cell,and interfere cell growth and cell cycle effectually,combining siRNA transfected was better than separate siRNA transfected in interfering effects to NPC CNE2 cell.This study provides new thoughts and methods for NPC gene therapy.
引文
1 闵华庆,洪明晃 郭翔等.鼻咽癌的研究进展.中国肿瘤,1999,8(12):558-560.
2 Zhou H D,Li G Y,Yang Y X,et al.Intracellular co-localization of SPLUNCl protein with nanobacteria in nasopharyngeal carcinoma epithelia HNEl cells depended on the bactericidal permeability increasing protein domain.Mol Immunol,2006,43(11):1864-1871.
3 黄麟波,陈德林,张锦明等.中国南北方鼻咽癌发病危险因素对比研究.癌症,1997,16(5):324-327.
4 Tsao SW,Tramoutanis G,Dawson CW,et all.The significance of LMPl exp- ression in nasopharyngeal carcinomal.Semin Cancer Biol,2002,12:473-487.
5 杨玉成 洪苏玲综述.EB病毒潜伏膜蛋白1与鼻咽癌.国际耳鼻咽喉头颈外科杂志,2007,31(1)19-22.
6 杨成勇,蔡伟民,沈倍奋等.鼻咽癌患者EB病毒潜伏膜蛋白(LMP- 1)的特异性细胞免疫研究.[J]病毒学报,1999,15(3):193-198.
7 Sheen TS,Huang YT,Chang YL,et all.Epstein-Barr virus- encoded latent membrane protein 1 co-expresses with epidermal growth factor receptor in nasopharyngeal carcinomal Jpn J Cancer Res,1999,90:1285-1292.
8 Hu LF,Troyanovasky B,Zhang XN,et al.Differences in the immunogenicity of latent membrane protein 1(LMP1) encoded by Epstein-Barr virus genomes derived from LMPl-positie and-negative nasopharyngeal carcinoma.Cancer Res,2000,60:5111-5116.
9 Vauxm DL,Coxy's Adamns JM.Bcl-2 gene promotes haemopoietic cell survival and cooperates with c-myc to immortalize pre-B cell.[J]Nature,1988,335(6189):440.
10 Cory S,Huang DC,Adams JM.The Bcl-2 family:roles in cell survival and oncogenesis[J].Oncogene,2003,22(53):8590-8607.
11 王晖,彭莉,涂青松等.鼻咽癌组织Bcl-2和Bax蛋白表达及其临床意义[J]. 中国医师杂志,2005,7(4):556-557.
12 胡利富,蒋金基,李晓林等.人鼻咽癌癌基因Ha-ras(J).癌症,1988,7(4):251.
13 蔡俊宏 综述,符生苗,邓立群 审校.鼻咽癌基因研究进展.中国热带医学.2004,4(1),131-134.
14 Sirotkovic- Skerlev M,Kirzanac S,Kapi tanovic S,et al.Expression of c - myc,erbB - 2,p53 and nm23 - H1 gene product in benign and malignant breast lesions:coexpression and correlation with clinicopathologic parameters[J].Exp Mol Pathol,2005,79(1):42 - 50.
15 Sheu F L,Chen A,Tseng H H,et al.Expression of p53 protein in nasopharyngeal carcinoma.J Pathol,1994,172:294-296,
16 Rubio L,Burgos JS,Lopez- Guerrero JA,et al.Expression of p53protein and tumorangiogenesis as prognostic factors in nasopharyngeal carcinoma patients[J].Pathol Res Pract,2002,198(2):97 - 102.
17 姜武忠,卢景琛,文继舫,等.鼻咽癌组织中nm23-H1和survivin蛋白表达及其临床意义[J].中华放射肿瘤学杂志,2005,14(5):383-387.
18 马志超,符生苗,蔡俊宏.RTQ-PCR检测鼻咽癌组织中nm23-H1基因表达研究.现代肿瘤医学,2008,16(1):17-19.
19 李华川综述,陆士新审校.抑癌基因研究进展.国外医学肿瘤学分册,1994,21(1)1-5.
20 黄光琦,宋毅,黄韧敏等.鼻咽癌Rb基因存在的状况研究.中华医学遗传学杂志,1994,11(2):75-77.
21 Sun Y,Hegamyer G,Colburn NH,et al.Nasopharyngeal carcinoma shows no detectable retinoblastoma susceptibility gene alterations.Oncogene,1993,791-793.
22 Lin CT,Chan W Y,Chen W,et al.Nasopharyngeal carcinoma and retinoblastoma gene expression.Lab Invest,1992,67(1) 56-58.
23 区宝祥.八株鼻咽癌细胞株的Rb基因及其表达.中华医学遗传学杂志,1997,14(6):368-370.
24 陈琦,张熔熔,王利军.p16和Rb蛋白在鼻咽癌中的表达及其意义.江苏医药,2002,28(9)706-707.
25 Morin GB.The human telomere terminal transferasa enzyme is a ribonucleoprotein that synthesizes TTAGGG repeat,cell,1989,59:521.
26 Orice CR.relomere structure in Enplotes crassus:characta rization of DNA- protein interactions and isolation of a telomere - binding protein.Mol Biol Cell,1990,10:3421.
27 Raghuraman MK,Cech TR.Effect of monovalent cation - induced telomeric DNA structure on the binding of Dxytricha telomeric protein.Nuleic Acids Res,1990,18:4543.
28 Takakura M,Kyo S,Kanaya T,etal;Expression of human telomerase subunits and correlation with telomerase activity in cervical cancer.(J),Cancer Res.1998,58:1558-1561.
29 王行炜,肖健云,赵素萍,等.鼻咽癌组织端粒酶各组分基因表达的研究[J].癌症,2001,20(3):245-248.
30 李登举综述 张瑶珍 何善述 审阅.端粒酶与细胞周期、细胞凋亡及癌基因的关系.国外医学分子生物学分册,2001,23(2)86-88.
31 Sumida T,Sogawa K,Hamakawa H,et al.Detection of telomerase activity in oral lesions.J Oral Pathol Med,1998,27(3):111-115.
32 Cheng RY,Yuen PW,Nicholls JM,et al.Telomerase activation in nasopharyngeal carcinomas.Br J Cancer,1998,77(3):456-460.
33 冯云,张小伯,李原,等.早期喉癌人乳头状瘤病毒感染和p53、p16、nm23蛋白表达及端粒酶激活相关性分析[J].中日友好医院学报,2004,18(3):158-162.
34 K.Miyachi,M.J.Fritzler and E.M.Tan,Autoantibody to a nuclear antigen in proliferating cells,J Immunol,1978,121(6):2228 - 2234.
35 Michaela Merkerova,Hana Bruchova and Radim Brdicka.Expression analysis of PCNA gene in chronic myelogenous leukemia-ombined application of siRNA silencing and expression arrays.Leukemia Research, 2007,31(5):661-672.
36 伍海军,申良方,肖华平等.增殖细胞核抗原和p16蛋白表达与鼻咽癌放射敏感性相关性研究.中国医师杂志,2007,9(5):619-621.
37 邹继彬,黄广恩,陈中坚等.Survivin蛋白和PCNA在鼻咽癌中的表达及其临床意义.中国医师杂志,2004,6(12):1636-1637.
38 Gorgoulis V,Zoumpourlis V,Rassidakis G,et al.Molecular analysis of p53 gene in laryngeal premalignant and malignant lesions,p53 protein immunohistochemical expression is positively related to proliferating cell nuclear antigen labeling index.Virchows Arch,1995,426:339-344.
39 Niemhom S,Kitazawa S,Murao S,et al.Co-expression of p53 and bcl-2may correlate to the presence of Epstein-barr virus genome and expression of proliferating cell nuclear antigen in nasopharyngeal carcinoma.Cancer Lett,2000,160:199-208.
40 赵恩宏,金小平,肖丽君等.端粒酶与增殖细胞核抗原在妊娠滋养细胞肿瘤中表达的研究.河北医药,2008,30(7):926-928.
41 林怀雪,胡德恩.鼻咽癌眼征171例分析[J].实用眼科杂志,1988,6(1):46-47.
42 谢民强,陶正德,肖健云等.误诊鼻咽癌50例分析,临床误诊误治1994,7(5):200-201.
43 李敏娥,章剑东.早期以眼部症状为主的鼻咽癌12例漏诊分析.浙江临床医学,2007,9(1):10-12.
44 张浩,颜建华,吴中耀等.鼻咽癌眼眶转移临床分析.中华眼科杂志,2006,42(4):318-322.
45 梁少波,刘立志,毛燕萍.鼻咽癌三叉神经侵犯的MRI评价及预后价值.中国肿瘤临床,2008,35(7),361-365.
46 龙晚生,王立,罗学毛.鼻咽癌眼眶浸润的临床及MRI诊断.中华眼科杂志,2001,37(4):295-297.
47 J.P.Spanoa,P.Bussonb,D.Atlanc.Nasopharyngeal carcinomas:an update,Review.European Journal of Cancer.2003,39:2121 - 2135.
48 Yupa Sumitsawan,Saisawat Chaiyasate,Imjai Chitapanarux.Late complications of radiotherapy for nasopharyngeal carcinoma.Auris Nasus Larynx.2009,36:205 -209.
49 孔琳,张有望.鼻咽癌放射治疗后神经系统后遗症.中华放射肿瘤学杂志1999年9月第8卷第3期181-183.
50 Rosenblatt E;Brook OR;Erlich N;et al.Late visual and auditory toxicity of radiotherapy for nasopharyngeal carcinoma.Tumori,2003,89(1):68-74.
51 张庆平,谢成熹,阮林等.鼻咽癌放疗性角膜损伤的跟踪研究.实用癌症杂志2001年11月第16卷第6期644-646.
52 姚照华,白志刚。鼻咽癌放疗后眼部损伤.伤残医学,1997,5(4):37-39.
53 李梅,欧杰雄,罗伟等.鼻咽癌放疗后放射性视网膜病变的临床分析.1999,17(12):731-732.
54 Peter M.L.Teoa,Sing Fai Leunga,Stewart Y.Tung et al.Dose - response relationship of nasopharyngeal carcinoma above conventional tumoricidal level:A study by the Hong Kong nasopharyngeal carcinoma study group (HKNPCSG)*.Radiotherapy and Oncology,2006,79:27- 33.
55 Lee AWM,Law SCK,Chan DKK,et al.Retrospective analysis of nasopharyngeal carcinoma treated during 1976-1985:late complications following megavoltage irradiation.Br J Radiol,1992,65(778):918-928.
56 Zhang QP,Ye CH,Huang GW,Xie CX,Zhou WW,Ruan L.A follow-up study on keratopathy in eyes after radiation therapy for nasopharyngeal carcinoma.Int J Ophthalmol(Guoji Yanke Zazhi),2006;6(4):755-757.
57 周平,王绍丰,苗重会.鼻咽癌放疗后并发视神经视网膜病变的临床研究.中华眼科杂志,2003,39(10):616-620.
58 Khochbin S et al.反义基因治疗文献(英文).Ann N Y Acad Sci,1992:660:77-88.
59 姚红兵,曹前,吴家廉.反义寡核苷酸对鼻咽癌细胞端粒酶活性的抑制.重庆医学,2003,32(4):481-482.
60 闵玲,周克元,梁统等.Bcl-xl反义寡核苷酸对裸鼠人鼻咽癌移植瘤抑制作用 的研究[J].中华肿瘤杂志,2004,26(1):14-17.
61 刘振声,李宝民,刘彦仿等.EB病毒反义LMP1基因对人低分化鼻咽癌CN E-3细胞株生长的抑制作用.中华耳鼻咽喉科杂志,1996,31:92-95.
62.W u CJ,L ue CY,L iu ST.et al.LMP1 as a selective therape uticstrategy for EBV-associated diseases.Gene Therapy,1998,5:905- 912.
63 黄海,周江红,潘星华等.BHRF1反义寡核苷酸对BHRF12CN E2细胞生存能力的影响.第二军医大学学报,1999,20(10):720-722.
64 Li JH,Li P,Klamut H,et al.Cytotoxic effects of Ad p53 expressiin two human nasopharyngeal carcinoma cell lines.Clin2 Cancer Re 1997,3(4):507-514.
65 W u CJ,L ue CY,L iu ST.et al,Transcriptional activation of NF-kB activity by Epstein-Barr virus(EBV)]。
66 Guo S,Kemhues KJ.Par-1,a gene required for establishing polarity in C.elegans embryos,encodes a putative Ser/Thr kinase that is asymmetrically distributed(J).Cell,1995,81(4):611-620.
67 Fire A,Xu S,Montgomery MK,et al.Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans.Nature,1998,391(6669):806-811.
68 DZ1TOYEVA S,DIM1TRUEVICN,MANI Ⅳ H.Intra-abdominal injection Of double-stranded RAN into anesthetized adult drosophila triggers RNA interference in the central nervous system.(J)Mol Psychiatry,2001,6(6):665-670.
69 Lipardi C,Wei Q,Paterson B M.RNAi as random degradtive PCR.siRNA primer convert mRNA into dsRNA that a degraded to generate new siRNAs[J],cell 2001,107:297-307.
70 Fire A.RNA-treggered gene siliencing[J].Trends Genet,1999,15(9):358-363.
71 Michael T.McManus and Phillip A.Sharp REVIEW.Gene silencing in mammals by small interfering RNAs.Nat Rev Genet 2003(10):737-747.
72 白爱民,贾艳敏,张晓菁.siRNA为基础的RNAi在肿瘤基因治疗中的应用进展.中华肿瘤防治杂志,2007,14(12):953-955.
73 张其亮,杨述华,刘红云等.增殖细胞核抗原短发夹状RNA对人骨肉瘤细胞生长的影响.中华病理学杂志,2005,34(3):167-170.
74 Li XP,Li G,Peng Y,et al.Suppression of Epstein-Barr virus encoded latent membrane protein-1 by RNA interference inhibits the metastatic potential of nasopharyngeal carcinoma cells.Biochem Biophys Res Commun,2004,315:212-218.
75 Borkhardt A.Blocking oncogenes in malignant cells by RNA interference-new hope for a highly specific cancer treatment?Cancer Cell,2002,2:167-168.
76 Ying RS,Zhu C,Fan XG,et al.Hepatitis B virus is inhibited by RNA interference in cell culture and in mice.
77 Elbashir,SM;Lendeckel,W;Tuschl T.RNA interference is mediated by 21-and 22-nucleotide RNAs.Journal,GENE DEVELOP JAN,2001,15:188-200.
78 房宝英,何冬梅,张洹.siRNA及其导入体内外方法的研究进展.国际病理科学与临床杂志.2007,27(1):44-47.
79 Li J X,Zhou KY,Cai KR,et al.Knockdown of bcl-xL expression with RNA interference induces nasophar yngeal carcinoma cells apoptosis[J].Chinese Journal of Otorhinolaryngology Head and Neck Surgery,2005,40(5):347-351.
80 Liu LZ,Liang CY,Dou YL,et al.Specific inhibition of Raf-1 gene expression in HNE1 by RNA interference[J].Chinese Journal of Otorhinolaryngology Head and Neck Surgery,2006,41(11):805-808.
81 李刚,李湘平,彭英等.小干扰RNA抑制EB病毒潜伏膜蛋白1表达对鼻咽癌细胞生长的影响.中华耳鼻咽喉头颈外科杂志,2005,40(6):406-410.
82 贺楚峰,赵素萍,肖健云,等.小干扰RNA抑制鼻咽癌细胞hTERT基因表达的实验研究.中国耳鼻咽喉颅底外科杂志2006,12(2):91-95.
83 沈永忠,王燕,陈始明等.短发夹RNA抑制hTERT基因对鼻咽癌细胞端粒酶活 性及PCNA和Caspase23蛋白表达的影响.临床耳鼻咽喉头颈外科杂志,2008,22(9):411-415.
84 SARETZKI G.Telomerase inhibition as cancer therapy[J].Cancer Letters,2003,194:209 - 219.
85 高国风,王沙燕,陈善义等.血管内皮生长因子RNA干扰效应对鼻咽癌生长的影响.广东医学,2006,27(6):785-787.
86 周海波,伍钢,彭纲.siRNA抑制VEGF基因表达对鼻咽癌细胞生物学行为的影响.临床肿瘤学杂志,2008,13(6):485-490.
87 翁德胜,吴正蓉,王爽等.RNAi抑制鼻咽癌细胞表皮生长受体表达对细胞生长的影响[J].南方医科大学报,2006,26(1):71-74.
1 卢泰祥,李家尧,胡永红,等.鼻咽癌颅底骨破坏的临床研究.中华耳鼻喉科杂志,1994,29:295-298.
2 杨立新,廉养德,王玖华.鼻咽癌咽旁间隙侵犯的CT诊断评价.临床放射学杂志,1998,17:23-25.
3 朱兰才,卢泰祥,张恩罴.鼻咽癌咽旁侵犯的预后探讨.癌症,1997,16:283-285.
4 Geara FB,Sanguineti G,Tucker SL,et al.Carcinoma of the nasopharynx treated by radiotherapy alone:determinants of distant metastasis and survival.RadiotherOncol,1997,43:53-61.
5 宋谦,刘国龙,盛信秀等.鼻咽癌远处转移的临床分析.临床肿瘤学杂志,1997.3:23-24.
6 谢民强 陶正德 肖健云等.误诊鼻咽癌50例分析,临床误诊误治1994,7(5),200-201.
7 黄跃.鼻咽癌在眼部表现14例临床分析.广西民族学院学报(自然科学版),2002,8(2):152-153.
8 Lee KY,Seah LL,Tow Set al.Nasopharyngeal carcinoma with orbital involvement.Ophthal Plast Reconstr Surg.2008 24(3),185-189.
9 龙晚生 王立 罗学毛.鼻咽癌眼眶浸润的临床及MRI诊断.中华眼科杂志,2001,37(4),295-298.
10 林怀雪,胡德恩。鼻咽癌眼症171例分析。中国实用眼科杂志,1988,6(1):46-47.
11 陈智聪,吴中耀,杨华胜等.28例眼眶转移癌的临床分析.中国实用眼科杂志,2002.20:837-840.
12 张浩 颜建华 吴中耀等,鼻咽癌眼眶转移病例分析.中华眼科杂志,2006,42(4),318-322.
13 梁少波 刘立志 毛燕萍.鼻咽癌三叉神经侵犯的MRI评价及预后价值.中国肿瘤临床,2008,35(7),361-365.
14 Zhang QP,Ye CH,Huang GW,Xie CX,Zhou WW,Ruan L.A follow-up study on keratopathy in eyes after radiation therapy for nasopharyngeal carcinoma.Int J Ophthalmol(Guoji Yanke Zazhi),2006;6(4):755-757.
15 周平,王绍丰,苗重会.鼻咽癌放疗后并发视神经视网膜病变的临床研究.中华眼科杂志,2003,39(10):616-620.
1 阿依恒·曲库尔汗,刘立中.端粒、端粒酶与头颈肿瘤.国际耳鼻咽喉头颈外科杂志,2007,31(1):1-3.
2 Satyanarayana A,Manns MP,Rudolph KL.Telomeres,telomerase and cancer:an endless search to target the ends,Cell Cycle,2004,3(9),1138-1150.
3 王行炜,肖健云,赵素萍,等.鼻咽癌组织端粒酶各组分基因表达的研究[J].癌症,2001,20(3):245-248.
4 Nimmo ER,Cranston G,Allshire RC.Telomere - associated chromosome breakage in fission yeast results in variegated expression of adjacent genes.EMBO J,1994,13:3801.
5 Gilson E,Laroche T,Gasser SM,et al.Telomeres and the functional architcsture of the nucleus.Trends Cell Biol,1993,3:128-130.
6 Morin GB.The human telomere terminal transferasa enzyme is a ribonucleoprotein that synthesizes TTAGGG repeat,cell,1989,59:521.
7 左连富主编.流式细胞术与生物医学.辽宁科学技术出版社,1996,32-68.
8 周汝麟,沈关心主编.现代免疫学实验技术.湖北科学技术出版社,1998,175-198.
9 Shiratsuchi M.Telomerase activity in myeloma cells is closely related to cell cycle status,but not to apoptotic signals induced by interferon-α,leuklymphoma 1999,34:349-359.
10 Vaziri H,Benchimol S.Alternative pathways for the extension of cellular life span;inactivation of p53/pRb an expression of telomerase.Oncogene.1999,18(53):7676-80.
11 陈意生,史景泉主编.肿瘤分子细胞生物学.第2版.北京人民军医出版社,2004,133-134.
1 程庆,魏晶晶,黄翔.增殖细胞核抗原和p53基因在鼻咽癌中的表达及相关性研究.临床耳鼻咽喉科杂志,2005,19(14):643-645.
2 石钦胜,黄致治,苏少雪.增殖细胞核抗原在鼻咽癌组织中的表达及其意义.癌症,1997,16(增刊):31-32.
3 钱金方.增殖细胞核抗原在常见肿瘤的研究进展.肿瘤基础与临床,2008,21(1):86-89.
4 Maga G,Hubscher U.Proliferating cell nuclear antigen(PCNA):A dancer with many partners.Journal of Cell Science,2003,116(15):3051-3060
5 宋楠萌 桑建利,徐恒 增殖细胞核抗原(PCNA)的分子结构及其生物学功能研究进展。自然科学进展。2006,16(10)1201-1209.
6 Vairapandi M,Azam N,Balliet A G,et al.Characterization of MyD118,Gadd45and proliferating cell nuclear antigen(PCNA) interacting domanins.The Journal of Biological Chemistry.2000,275(22):16810-16819.
7 Haupt S,Berger M,Goldgerg Z,et al.Apoptosis-p53 nerwork.Journal of Cell Science,2003,116(20):4077-4085.
8 Scottl M,Bonnefin P,Vieyra D,et al.UV-induced binding of ING1 to PCNA regulates the induction of apoposis.Journal of Cell Science,2001,114:3455-3462.
9 Fujise K,Zhang D,Liu J L,et al.Regulation of apoptosis and cell cycle progression by MCL1 differential role of proliferating cell nuclear antigen.The Journal of Biological Chemistry.2000,275(50):39458-39465.
10 Niemhom S,Kitazawa S,Murao S,et al.Co-expression of p53 and bcl-2 may correlate to the presence of Ep-stein-barr virus genome and expression of proliferating cell nuclear antigen in nasopharyngeal carcinoma.Cancer Lett,2000,160:199 - 208.
11 钱洪流,宗永生,张昌卿.鼻咽癌中p53蛋白积聚的生物学特征和临床意义.临床与实验病理学杂志,1996,12(1):15-18.
1 胡晓慧,范磊,阮长耿.RNA干扰法对内皮细胞基质金属蛋白酶-2功能的研究.中国实验血液学杂志,2008,16(2):381-386.
2 李桂源,刘华英,周鸣等.鼻咽癌癌变的分子机理.生物化学与生物物理进展.2006,33(10):922-931.
3 李登举综述;张瑶珍,何善述 审阅.端粒酶与细胞周期_细胞凋亡及癌基因的关系.国外医学分子生物学分册,2001,23(2):86-89.
4 宋楠萌 桑建利,徐恒,增殖细胞核抗原(PCNA)的分子结构及其生物学功能研究进展.自然科学进展,2006,16(10):1201-1209.
5 黄光琦,宋毅,黄韧敏等.鼻咽癌Rb基因存在的状况研究.中华医学遗传学杂志,1994,11(2):75-77.
6 区宝祥.八株鼻咽癌细胞株的Rb基因及其表达.中华医学遗传学杂志,1997,14(6):368-370.
7 Kerr JF,Wyllie AH,Currie AR.Apoptosis:a basic biological pheonmenon with wide-ranging implications in tissue kinetics.Br J Cancer.1972 Aug,26(4):239-257.
8 Feng X L,Hara Y,Riabowol K.Different HATS of the ING1 gene family.Trends in Cell Biology,2002,12:532.
9 Nicoletti Ⅰ,Migliorati G,Pagliacci M C.A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry.J Immunol Methods,1991;139:271.
10 赵恩宏 金小平 肖丽君.杨宗伟端粒酶与增殖细胞核抗原在妊娠滋养细胞肿瘤中表达的研究.河北医药,2008,30(7):926-928.
11 沈永忠,王燕,陈始明等.短发夹RNA抑制hTERT基因对鼻咽癌细胞端粒酶活性及PCNA和Caspase23蛋白表达的影响.临床耳鼻咽喉头颈外科杂志,2008,22(9):411-415.
12 周俊旭,周绪红,陶泽璋等.小干扰RNA对喉鳞状细胞癌裸鼠移植瘤增殖细胞核抗原及p53蛋白表达的影响.临床耳鼻咽喉科杂志,2006,20(6):264-267.
13 Ding L;Li L;Yang J,et al.Latent membrane protein 1 encoded by Epstein-Barr virus induces telomerase activity via p16INK4A/Rb/E2F1 and JNK signaling pathways.Journal Of Medical Virology[J Med Virol]2007 Aug;Vol.79(8):1153-63.
14 Michaela Merkerova,Hana Bruchova,Radim Brdicka.Expression analysis of PCNA gene in chronic myelogenous leukemia--Combined application of siRNA silencing and expression arrays.Leukemia Research,2007,31(5):661-672.
15 谢庆祥,林吓聪,韩聪祥.E2F-1在梗阻性肾组织中的表达及其意义.中国现代医学杂志,2007,17(7):871-873.
16 景抗震硕士论文,导师高建平.联合RNA干扰hTR及hTERT基因对膀胱尿路上皮癌BIU-87细胞株端粒酶活性抑制研究.第二军医大学,2008年3月.
2 Zhou H D,Li G Y,Yang Y X,et al.Intracellular co-localization of SPLUNCl protein with nanobacteria in nasopharyngeal carcinoma epithelia HNEl cells depended on the bactericidal permeability increasing protein domain.Mol Immunol,2006,43(11):1864-1871.
3 黄麟波,陈德林,张锦明等.中国南北方鼻咽癌发病危险因素对比研究.癌症,1997,16(5):324-327.
4 Tsao SW,Tramoutanis G,Dawson CW,et all.The significance of LMPl exp- ression in nasopharyngeal carcinomal.Semin Cancer Biol,2002,12:473-487.
5 杨玉成 洪苏玲综述.EB病毒潜伏膜蛋白1与鼻咽癌.国际耳鼻咽喉头颈外科杂志,2007,31(1)19-22.
6 杨成勇,蔡伟民,沈倍奋等.鼻咽癌患者EB病毒潜伏膜蛋白(LMP- 1)的特异性细胞免疫研究.[J]病毒学报,1999,15(3):193-198.
7 Sheen TS,Huang YT,Chang YL,et all.Epstein-Barr virus- encoded latent membrane protein 1 co-expresses with epidermal growth factor receptor in nasopharyngeal carcinomal Jpn J Cancer Res,1999,90:1285-1292.
8 Hu LF,Troyanovasky B,Zhang XN,et al.Differences in the immunogenicity of latent membrane protein 1(LMP1) encoded by Epstein-Barr virus genomes derived from LMPl-positie and-negative nasopharyngeal carcinoma.Cancer Res,2000,60:5111-5116.
9 Vauxm DL,Coxy's Adamns JM.Bcl-2 gene promotes haemopoietic cell survival and cooperates with c-myc to immortalize pre-B cell.[J]Nature,1988,335(6189):440.
10 Cory S,Huang DC,Adams JM.The Bcl-2 family:roles in cell survival and oncogenesis[J].Oncogene,2003,22(53):8590-8607.
11 王晖,彭莉,涂青松等.鼻咽癌组织Bcl-2和Bax蛋白表达及其临床意义[J]. 中国医师杂志,2005,7(4):556-557.
12 胡利富,蒋金基,李晓林等.人鼻咽癌癌基因Ha-ras(J).癌症,1988,7(4):251.
13 蔡俊宏 综述,符生苗,邓立群 审校.鼻咽癌基因研究进展.中国热带医学.2004,4(1),131-134.
14 Sirotkovic- Skerlev M,Kirzanac S,Kapi tanovic S,et al.Expression of c - myc,erbB - 2,p53 and nm23 - H1 gene product in benign and malignant breast lesions:coexpression and correlation with clinicopathologic parameters[J].Exp Mol Pathol,2005,79(1):42 - 50.
15 Sheu F L,Chen A,Tseng H H,et al.Expression of p53 protein in nasopharyngeal carcinoma.J Pathol,1994,172:294-296,
16 Rubio L,Burgos JS,Lopez- Guerrero JA,et al.Expression of p53protein and tumorangiogenesis as prognostic factors in nasopharyngeal carcinoma patients[J].Pathol Res Pract,2002,198(2):97 - 102.
17 姜武忠,卢景琛,文继舫,等.鼻咽癌组织中nm23-H1和survivin蛋白表达及其临床意义[J].中华放射肿瘤学杂志,2005,14(5):383-387.
18 马志超,符生苗,蔡俊宏.RTQ-PCR检测鼻咽癌组织中nm23-H1基因表达研究.现代肿瘤医学,2008,16(1):17-19.
19 李华川综述,陆士新审校.抑癌基因研究进展.国外医学肿瘤学分册,1994,21(1)1-5.
20 黄光琦,宋毅,黄韧敏等.鼻咽癌Rb基因存在的状况研究.中华医学遗传学杂志,1994,11(2):75-77.
21 Sun Y,Hegamyer G,Colburn NH,et al.Nasopharyngeal carcinoma shows no detectable retinoblastoma susceptibility gene alterations.Oncogene,1993,791-793.
22 Lin CT,Chan W Y,Chen W,et al.Nasopharyngeal carcinoma and retinoblastoma gene expression.Lab Invest,1992,67(1) 56-58.
23 区宝祥.八株鼻咽癌细胞株的Rb基因及其表达.中华医学遗传学杂志,1997,14(6):368-370.
24 陈琦,张熔熔,王利军.p16和Rb蛋白在鼻咽癌中的表达及其意义.江苏医药,2002,28(9)706-707.
25 Morin GB.The human telomere terminal transferasa enzyme is a ribonucleoprotein that synthesizes TTAGGG repeat,cell,1989,59:521.
26 Orice CR.relomere structure in Enplotes crassus:characta rization of DNA- protein interactions and isolation of a telomere - binding protein.Mol Biol Cell,1990,10:3421.
27 Raghuraman MK,Cech TR.Effect of monovalent cation - induced telomeric DNA structure on the binding of Dxytricha telomeric protein.Nuleic Acids Res,1990,18:4543.
28 Takakura M,Kyo S,Kanaya T,etal;Expression of human telomerase subunits and correlation with telomerase activity in cervical cancer.(J),Cancer Res.1998,58:1558-1561.
29 王行炜,肖健云,赵素萍,等.鼻咽癌组织端粒酶各组分基因表达的研究[J].癌症,2001,20(3):245-248.
30 李登举综述 张瑶珍 何善述 审阅.端粒酶与细胞周期、细胞凋亡及癌基因的关系.国外医学分子生物学分册,2001,23(2)86-88.
31 Sumida T,Sogawa K,Hamakawa H,et al.Detection of telomerase activity in oral lesions.J Oral Pathol Med,1998,27(3):111-115.
32 Cheng RY,Yuen PW,Nicholls JM,et al.Telomerase activation in nasopharyngeal carcinomas.Br J Cancer,1998,77(3):456-460.
33 冯云,张小伯,李原,等.早期喉癌人乳头状瘤病毒感染和p53、p16、nm23蛋白表达及端粒酶激活相关性分析[J].中日友好医院学报,2004,18(3):158-162.
34 K.Miyachi,M.J.Fritzler and E.M.Tan,Autoantibody to a nuclear antigen in proliferating cells,J Immunol,1978,121(6):2228 - 2234.
35 Michaela Merkerova,Hana Bruchova and Radim Brdicka.Expression analysis of PCNA gene in chronic myelogenous leukemia-ombined application of siRNA silencing and expression arrays.Leukemia Research, 2007,31(5):661-672.
36 伍海军,申良方,肖华平等.增殖细胞核抗原和p16蛋白表达与鼻咽癌放射敏感性相关性研究.中国医师杂志,2007,9(5):619-621.
37 邹继彬,黄广恩,陈中坚等.Survivin蛋白和PCNA在鼻咽癌中的表达及其临床意义.中国医师杂志,2004,6(12):1636-1637.
38 Gorgoulis V,Zoumpourlis V,Rassidakis G,et al.Molecular analysis of p53 gene in laryngeal premalignant and malignant lesions,p53 protein immunohistochemical expression is positively related to proliferating cell nuclear antigen labeling index.Virchows Arch,1995,426:339-344.
39 Niemhom S,Kitazawa S,Murao S,et al.Co-expression of p53 and bcl-2may correlate to the presence of Epstein-barr virus genome and expression of proliferating cell nuclear antigen in nasopharyngeal carcinoma.Cancer Lett,2000,160:199-208.
40 赵恩宏,金小平,肖丽君等.端粒酶与增殖细胞核抗原在妊娠滋养细胞肿瘤中表达的研究.河北医药,2008,30(7):926-928.
41 林怀雪,胡德恩.鼻咽癌眼征171例分析[J].实用眼科杂志,1988,6(1):46-47.
42 谢民强,陶正德,肖健云等.误诊鼻咽癌50例分析,临床误诊误治1994,7(5):200-201.
43 李敏娥,章剑东.早期以眼部症状为主的鼻咽癌12例漏诊分析.浙江临床医学,2007,9(1):10-12.
44 张浩,颜建华,吴中耀等.鼻咽癌眼眶转移临床分析.中华眼科杂志,2006,42(4):318-322.
45 梁少波,刘立志,毛燕萍.鼻咽癌三叉神经侵犯的MRI评价及预后价值.中国肿瘤临床,2008,35(7),361-365.
46 龙晚生,王立,罗学毛.鼻咽癌眼眶浸润的临床及MRI诊断.中华眼科杂志,2001,37(4):295-297.
47 J.P.Spanoa,P.Bussonb,D.Atlanc.Nasopharyngeal carcinomas:an update,Review.European Journal of Cancer.2003,39:2121 - 2135.
48 Yupa Sumitsawan,Saisawat Chaiyasate,Imjai Chitapanarux.Late complications of radiotherapy for nasopharyngeal carcinoma.Auris Nasus Larynx.2009,36:205 -209.
49 孔琳,张有望.鼻咽癌放射治疗后神经系统后遗症.中华放射肿瘤学杂志1999年9月第8卷第3期181-183.
50 Rosenblatt E;Brook OR;Erlich N;et al.Late visual and auditory toxicity of radiotherapy for nasopharyngeal carcinoma.Tumori,2003,89(1):68-74.
51 张庆平,谢成熹,阮林等.鼻咽癌放疗性角膜损伤的跟踪研究.实用癌症杂志2001年11月第16卷第6期644-646.
52 姚照华,白志刚。鼻咽癌放疗后眼部损伤.伤残医学,1997,5(4):37-39.
53 李梅,欧杰雄,罗伟等.鼻咽癌放疗后放射性视网膜病变的临床分析.1999,17(12):731-732.
54 Peter M.L.Teoa,Sing Fai Leunga,Stewart Y.Tung et al.Dose - response relationship of nasopharyngeal carcinoma above conventional tumoricidal level:A study by the Hong Kong nasopharyngeal carcinoma study group (HKNPCSG)*.Radiotherapy and Oncology,2006,79:27- 33.
55 Lee AWM,Law SCK,Chan DKK,et al.Retrospective analysis of nasopharyngeal carcinoma treated during 1976-1985:late complications following megavoltage irradiation.Br J Radiol,1992,65(778):918-928.
56 Zhang QP,Ye CH,Huang GW,Xie CX,Zhou WW,Ruan L.A follow-up study on keratopathy in eyes after radiation therapy for nasopharyngeal carcinoma.Int J Ophthalmol(Guoji Yanke Zazhi),2006;6(4):755-757.
57 周平,王绍丰,苗重会.鼻咽癌放疗后并发视神经视网膜病变的临床研究.中华眼科杂志,2003,39(10):616-620.
58 Khochbin S et al.反义基因治疗文献(英文).Ann N Y Acad Sci,1992:660:77-88.
59 姚红兵,曹前,吴家廉.反义寡核苷酸对鼻咽癌细胞端粒酶活性的抑制.重庆医学,2003,32(4):481-482.
60 闵玲,周克元,梁统等.Bcl-xl反义寡核苷酸对裸鼠人鼻咽癌移植瘤抑制作用 的研究[J].中华肿瘤杂志,2004,26(1):14-17.
61 刘振声,李宝民,刘彦仿等.EB病毒反义LMP1基因对人低分化鼻咽癌CN E-3细胞株生长的抑制作用.中华耳鼻咽喉科杂志,1996,31:92-95.
62.W u CJ,L ue CY,L iu ST.et al.LMP1 as a selective therape uticstrategy for EBV-associated diseases.Gene Therapy,1998,5:905- 912.
63 黄海,周江红,潘星华等.BHRF1反义寡核苷酸对BHRF12CN E2细胞生存能力的影响.第二军医大学学报,1999,20(10):720-722.
64 Li JH,Li P,Klamut H,et al.Cytotoxic effects of Ad p53 expressiin two human nasopharyngeal carcinoma cell lines.Clin2 Cancer Re 1997,3(4):507-514.
65 W u CJ,L ue CY,L iu ST.et al,Transcriptional activation of NF-kB activity by Epstein-Barr virus(EBV)]。
66 Guo S,Kemhues KJ.Par-1,a gene required for establishing polarity in C.elegans embryos,encodes a putative Ser/Thr kinase that is asymmetrically distributed(J).Cell,1995,81(4):611-620.
67 Fire A,Xu S,Montgomery MK,et al.Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans.Nature,1998,391(6669):806-811.
68 DZ1TOYEVA S,DIM1TRUEVICN,MANI Ⅳ H.Intra-abdominal injection Of double-stranded RAN into anesthetized adult drosophila triggers RNA interference in the central nervous system.(J)Mol Psychiatry,2001,6(6):665-670.
69 Lipardi C,Wei Q,Paterson B M.RNAi as random degradtive PCR.siRNA primer convert mRNA into dsRNA that a degraded to generate new siRNAs[J],cell 2001,107:297-307.
70 Fire A.RNA-treggered gene siliencing[J].Trends Genet,1999,15(9):358-363.
71 Michael T.McManus and Phillip A.Sharp REVIEW.Gene silencing in mammals by small interfering RNAs.Nat Rev Genet 2003(10):737-747.
72 白爱民,贾艳敏,张晓菁.siRNA为基础的RNAi在肿瘤基因治疗中的应用进展.中华肿瘤防治杂志,2007,14(12):953-955.
73 张其亮,杨述华,刘红云等.增殖细胞核抗原短发夹状RNA对人骨肉瘤细胞生长的影响.中华病理学杂志,2005,34(3):167-170.
74 Li XP,Li G,Peng Y,et al.Suppression of Epstein-Barr virus encoded latent membrane protein-1 by RNA interference inhibits the metastatic potential of nasopharyngeal carcinoma cells.Biochem Biophys Res Commun,2004,315:212-218.
75 Borkhardt A.Blocking oncogenes in malignant cells by RNA interference-new hope for a highly specific cancer treatment?Cancer Cell,2002,2:167-168.
76 Ying RS,Zhu C,Fan XG,et al.Hepatitis B virus is inhibited by RNA interference in cell culture and in mice.
77 Elbashir,SM;Lendeckel,W;Tuschl T.RNA interference is mediated by 21-and 22-nucleotide RNAs.Journal,GENE DEVELOP JAN,2001,15:188-200.
78 房宝英,何冬梅,张洹.siRNA及其导入体内外方法的研究进展.国际病理科学与临床杂志.2007,27(1):44-47.
79 Li J X,Zhou KY,Cai KR,et al.Knockdown of bcl-xL expression with RNA interference induces nasophar yngeal carcinoma cells apoptosis[J].Chinese Journal of Otorhinolaryngology Head and Neck Surgery,2005,40(5):347-351.
80 Liu LZ,Liang CY,Dou YL,et al.Specific inhibition of Raf-1 gene expression in HNE1 by RNA interference[J].Chinese Journal of Otorhinolaryngology Head and Neck Surgery,2006,41(11):805-808.
81 李刚,李湘平,彭英等.小干扰RNA抑制EB病毒潜伏膜蛋白1表达对鼻咽癌细胞生长的影响.中华耳鼻咽喉头颈外科杂志,2005,40(6):406-410.
82 贺楚峰,赵素萍,肖健云,等.小干扰RNA抑制鼻咽癌细胞hTERT基因表达的实验研究.中国耳鼻咽喉颅底外科杂志2006,12(2):91-95.
83 沈永忠,王燕,陈始明等.短发夹RNA抑制hTERT基因对鼻咽癌细胞端粒酶活 性及PCNA和Caspase23蛋白表达的影响.临床耳鼻咽喉头颈外科杂志,2008,22(9):411-415.
84 SARETZKI G.Telomerase inhibition as cancer therapy[J].Cancer Letters,2003,194:209 - 219.
85 高国风,王沙燕,陈善义等.血管内皮生长因子RNA干扰效应对鼻咽癌生长的影响.广东医学,2006,27(6):785-787.
86 周海波,伍钢,彭纲.siRNA抑制VEGF基因表达对鼻咽癌细胞生物学行为的影响.临床肿瘤学杂志,2008,13(6):485-490.
87 翁德胜,吴正蓉,王爽等.RNAi抑制鼻咽癌细胞表皮生长受体表达对细胞生长的影响[J].南方医科大学报,2006,26(1):71-74.
1 卢泰祥,李家尧,胡永红,等.鼻咽癌颅底骨破坏的临床研究.中华耳鼻喉科杂志,1994,29:295-298.
2 杨立新,廉养德,王玖华.鼻咽癌咽旁间隙侵犯的CT诊断评价.临床放射学杂志,1998,17:23-25.
3 朱兰才,卢泰祥,张恩罴.鼻咽癌咽旁侵犯的预后探讨.癌症,1997,16:283-285.
4 Geara FB,Sanguineti G,Tucker SL,et al.Carcinoma of the nasopharynx treated by radiotherapy alone:determinants of distant metastasis and survival.RadiotherOncol,1997,43:53-61.
5 宋谦,刘国龙,盛信秀等.鼻咽癌远处转移的临床分析.临床肿瘤学杂志,1997.3:23-24.
6 谢民强 陶正德 肖健云等.误诊鼻咽癌50例分析,临床误诊误治1994,7(5),200-201.
7 黄跃.鼻咽癌在眼部表现14例临床分析.广西民族学院学报(自然科学版),2002,8(2):152-153.
8 Lee KY,Seah LL,Tow Set al.Nasopharyngeal carcinoma with orbital involvement.Ophthal Plast Reconstr Surg.2008 24(3),185-189.
9 龙晚生 王立 罗学毛.鼻咽癌眼眶浸润的临床及MRI诊断.中华眼科杂志,2001,37(4),295-298.
10 林怀雪,胡德恩。鼻咽癌眼症171例分析。中国实用眼科杂志,1988,6(1):46-47.
11 陈智聪,吴中耀,杨华胜等.28例眼眶转移癌的临床分析.中国实用眼科杂志,2002.20:837-840.
12 张浩 颜建华 吴中耀等,鼻咽癌眼眶转移病例分析.中华眼科杂志,2006,42(4),318-322.
13 梁少波 刘立志 毛燕萍.鼻咽癌三叉神经侵犯的MRI评价及预后价值.中国肿瘤临床,2008,35(7),361-365.
14 Zhang QP,Ye CH,Huang GW,Xie CX,Zhou WW,Ruan L.A follow-up study on keratopathy in eyes after radiation therapy for nasopharyngeal carcinoma.Int J Ophthalmol(Guoji Yanke Zazhi),2006;6(4):755-757.
15 周平,王绍丰,苗重会.鼻咽癌放疗后并发视神经视网膜病变的临床研究.中华眼科杂志,2003,39(10):616-620.
1 阿依恒·曲库尔汗,刘立中.端粒、端粒酶与头颈肿瘤.国际耳鼻咽喉头颈外科杂志,2007,31(1):1-3.
2 Satyanarayana A,Manns MP,Rudolph KL.Telomeres,telomerase and cancer:an endless search to target the ends,Cell Cycle,2004,3(9),1138-1150.
3 王行炜,肖健云,赵素萍,等.鼻咽癌组织端粒酶各组分基因表达的研究[J].癌症,2001,20(3):245-248.
4 Nimmo ER,Cranston G,Allshire RC.Telomere - associated chromosome breakage in fission yeast results in variegated expression of adjacent genes.EMBO J,1994,13:3801.
5 Gilson E,Laroche T,Gasser SM,et al.Telomeres and the functional architcsture of the nucleus.Trends Cell Biol,1993,3:128-130.
6 Morin GB.The human telomere terminal transferasa enzyme is a ribonucleoprotein that synthesizes TTAGGG repeat,cell,1989,59:521.
7 左连富主编.流式细胞术与生物医学.辽宁科学技术出版社,1996,32-68.
8 周汝麟,沈关心主编.现代免疫学实验技术.湖北科学技术出版社,1998,175-198.
9 Shiratsuchi M.Telomerase activity in myeloma cells is closely related to cell cycle status,but not to apoptotic signals induced by interferon-α,leuklymphoma 1999,34:349-359.
10 Vaziri H,Benchimol S.Alternative pathways for the extension of cellular life span;inactivation of p53/pRb an expression of telomerase.Oncogene.1999,18(53):7676-80.
11 陈意生,史景泉主编.肿瘤分子细胞生物学.第2版.北京人民军医出版社,2004,133-134.
1 程庆,魏晶晶,黄翔.增殖细胞核抗原和p53基因在鼻咽癌中的表达及相关性研究.临床耳鼻咽喉科杂志,2005,19(14):643-645.
2 石钦胜,黄致治,苏少雪.增殖细胞核抗原在鼻咽癌组织中的表达及其意义.癌症,1997,16(增刊):31-32.
3 钱金方.增殖细胞核抗原在常见肿瘤的研究进展.肿瘤基础与临床,2008,21(1):86-89.
4 Maga G,Hubscher U.Proliferating cell nuclear antigen(PCNA):A dancer with many partners.Journal of Cell Science,2003,116(15):3051-3060
5 宋楠萌 桑建利,徐恒 增殖细胞核抗原(PCNA)的分子结构及其生物学功能研究进展。自然科学进展。2006,16(10)1201-1209.
6 Vairapandi M,Azam N,Balliet A G,et al.Characterization of MyD118,Gadd45and proliferating cell nuclear antigen(PCNA) interacting domanins.The Journal of Biological Chemistry.2000,275(22):16810-16819.
7 Haupt S,Berger M,Goldgerg Z,et al.Apoptosis-p53 nerwork.Journal of Cell Science,2003,116(20):4077-4085.
8 Scottl M,Bonnefin P,Vieyra D,et al.UV-induced binding of ING1 to PCNA regulates the induction of apoposis.Journal of Cell Science,2001,114:3455-3462.
9 Fujise K,Zhang D,Liu J L,et al.Regulation of apoptosis and cell cycle progression by MCL1 differential role of proliferating cell nuclear antigen.The Journal of Biological Chemistry.2000,275(50):39458-39465.
10 Niemhom S,Kitazawa S,Murao S,et al.Co-expression of p53 and bcl-2 may correlate to the presence of Ep-stein-barr virus genome and expression of proliferating cell nuclear antigen in nasopharyngeal carcinoma.Cancer Lett,2000,160:199 - 208.
11 钱洪流,宗永生,张昌卿.鼻咽癌中p53蛋白积聚的生物学特征和临床意义.临床与实验病理学杂志,1996,12(1):15-18.
1 胡晓慧,范磊,阮长耿.RNA干扰法对内皮细胞基质金属蛋白酶-2功能的研究.中国实验血液学杂志,2008,16(2):381-386.
2 李桂源,刘华英,周鸣等.鼻咽癌癌变的分子机理.生物化学与生物物理进展.2006,33(10):922-931.
3 李登举综述;张瑶珍,何善述 审阅.端粒酶与细胞周期_细胞凋亡及癌基因的关系.国外医学分子生物学分册,2001,23(2):86-89.
4 宋楠萌 桑建利,徐恒,增殖细胞核抗原(PCNA)的分子结构及其生物学功能研究进展.自然科学进展,2006,16(10):1201-1209.
5 黄光琦,宋毅,黄韧敏等.鼻咽癌Rb基因存在的状况研究.中华医学遗传学杂志,1994,11(2):75-77.
6 区宝祥.八株鼻咽癌细胞株的Rb基因及其表达.中华医学遗传学杂志,1997,14(6):368-370.
7 Kerr JF,Wyllie AH,Currie AR.Apoptosis:a basic biological pheonmenon with wide-ranging implications in tissue kinetics.Br J Cancer.1972 Aug,26(4):239-257.
8 Feng X L,Hara Y,Riabowol K.Different HATS of the ING1 gene family.Trends in Cell Biology,2002,12:532.
9 Nicoletti Ⅰ,Migliorati G,Pagliacci M C.A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry.J Immunol Methods,1991;139:271.
10 赵恩宏 金小平 肖丽君.杨宗伟端粒酶与增殖细胞核抗原在妊娠滋养细胞肿瘤中表达的研究.河北医药,2008,30(7):926-928.
11 沈永忠,王燕,陈始明等.短发夹RNA抑制hTERT基因对鼻咽癌细胞端粒酶活性及PCNA和Caspase23蛋白表达的影响.临床耳鼻咽喉头颈外科杂志,2008,22(9):411-415.
12 周俊旭,周绪红,陶泽璋等.小干扰RNA对喉鳞状细胞癌裸鼠移植瘤增殖细胞核抗原及p53蛋白表达的影响.临床耳鼻咽喉科杂志,2006,20(6):264-267.
13 Ding L;Li L;Yang J,et al.Latent membrane protein 1 encoded by Epstein-Barr virus induces telomerase activity via p16INK4A/Rb/E2F1 and JNK signaling pathways.Journal Of Medical Virology[J Med Virol]2007 Aug;Vol.79(8):1153-63.
14 Michaela Merkerova,Hana Bruchova,Radim Brdicka.Expression analysis of PCNA gene in chronic myelogenous leukemia--Combined application of siRNA silencing and expression arrays.Leukemia Research,2007,31(5):661-672.
15 谢庆祥,林吓聪,韩聪祥.E2F-1在梗阻性肾组织中的表达及其意义.中国现代医学杂志,2007,17(7):871-873.
16 景抗震硕士论文,导师高建平.联合RNA干扰hTR及hTERT基因对膀胱尿路上皮癌BIU-87细胞株端粒酶活性抑制研究.第二军医大学,2008年3月.