肺癌组织中ASPP基因家族启动子甲基化与p53凋亡通路相关性研究
|
摘要
目的：为阐明ASPP基因家族与肺癌(Lung carcinoma)发生发展的关系,本文从基因水平检测肺癌组织中抑癌基因ASPP1、ASPP2启动子甲基化状态；从蛋白水平上检测ASPP1、ASPP2、p53在肺癌组织中的表达情况,并检测肺癌组织的凋亡指数(apoptosis index,AI),进而分析其间的相关性,探讨ASPP家族表达调控机制及其在p53凋亡通路中的作用,为肺癌的早期诊断,治疗及预后判断提供理论依据。
方法：本研究实验组收集肺癌组织90例,对照组25例相应癌旁正常肺组织,构建肺癌组织芯片,免疫组织化学染色Envision法检测ASPP1、ASPP2及p53蛋白在肺癌组织芯片中的表达情况,并同时利用原位末端转移酶标记检测法(Terminal deoxynucleotidyl transferase dUTP nick end labeling,TUNEL法)检测肺癌细胞的凋亡指数AI,应用甲基化特异性PCR(methylation-specific PCR,MSP)对抑癌基因ASPP1、ASPP2启动子CpG岛进行甲基化状态检测,分析ASPP1、ASPP2蛋白表达、启动子甲基化和p53细胞凋亡通路之间的关系。
结果
1.90例肺癌组织ASPP1和ASPP2阳性率分别为27.8％和11.1％；25例癌旁组织中ASPP1和ASPP2阳性率分别为52.0％和16.0％；ASPP1在肺癌组织和癌旁组织中表达阳性率两者比较,差异有统计学意义(χ2=5.188,P<0.05),而ASPP2在肺癌组织和癌旁组织中表达阳性率两者比较,差异无统计学意义(χ2=0.437,P>0.05)。
2.90例肺癌组织中ASPP1蛋白的表达和分化程度、TNM分期及淋巴结转移呈正相关(P<0.05),与患者的性别、年龄、肿瘤大小、组织学类型无关(P>0.05)；而ASPP2蛋白的表达与肺癌的临床病理特征均无关(P>0.05)。
3.39例p53表达阴性的肺癌组织中ASPP1表达阳性16例(41.02％),51例突变型p53肺癌组织中ASPP1表达阳性9例(17.64％),经Spearman's相关分析,说明在肺癌组织中ASPP1蛋白表达与突变型p53蛋白表达呈负相关(r=-0.259,P<0.01),而39例p53表达阴性的肺癌患者中ASPP2蛋白表达阳性6例(15.38％),51例突变型p53中4例ASPP2表达阳性(9.75％),经Spearman's相关分析,可认为在肺癌组织中ASPP2蛋白表达与p53蛋白的表达无相关性(r=-0.119,P>0.05)。
4.比较90肺癌组织ASPP1蛋白阳性表达组与阴性组中AI值,发现ASPP1蛋白阳性组高于阴性组,提示ASPP1蛋白表达与细胞凋亡有关(t=2.337,P<0.05)；而90例肺癌组织中,ASPP2蛋白表达阳性组和阴性组之间AI值无显著性差异(t=1.083,P>0.05),可以为ASPP2蛋白的表达与肺癌细胞凋亡无关。
5.NSCLC中ASPP1基因启动子甲基化率为42.22％(38／90),癌旁组织甲基化率为16.00％(4／25),有显著性差异(χ2=5.830,P>0.05)；ASPP1基因启动子甲基化与患者的年龄、性别、组织类型和分化程度无关(χ2=1.129、1.488、1.126、4.555,P>0.05),但与患者的TNM分期和淋巴结转移有关(χ2=6.919、5.213,P=<0.05)；90例肺癌组织中ASPP1甲基化者其蛋白表达率明显低于未甲基化者(χ2=5.188,P<0.05)；而ASPP2基因启动子在90例NSCLC和25例癌旁组织中均未检测到甲基化状态。
结论
1.ASPP1蛋白在肺癌组织中低表达,与肿瘤分化程度、临床分期和淋巴结转移有关,提示ASPP1表达缺失可能与肺癌发生、进展有关。
2.ASPP2蛋白在肺癌组织和癌旁肺组织中的表达无明显差异,提示ASPP2在肺癌中可能不具有肿瘤抑制基因特性。
3.在肺癌组织中,ASPP1促进肺癌细胞AI的增高,且其表达与突变型p53呈负相关,ASPP1可能是p53凋亡通路中的关键蛋白；而ASPP2可能与p53凋亡通路无关。
4.肺癌组织中ASPP1启动子甲基化可能是ASPP1蛋白表达下调的原因,ASPP1高甲基化预示着NSCLC的高侵袭性,而ASPP2基因甲基化可能不是其蛋白的表达的调控机制。
Objective:In order to research the relation between ASPP gene family and incidence of lung carcinoma, the article examined the promoter methylation statut of the ASPP1 and ASPP2 from lever of DNA in lung carcinoma, and the expression of ASPP1、ASPP2 and p53 from lever of protein, and the apoptosis index of the lung cancer tissues.
Metholds:To construct tissue microarray of lung cancers,we collected 90 cases lung carcinoma tissues as experimental group and 25 normal adjacent lung tissue as control group. The expression state of ASPP1、ASPP2 and p53 protion were determined by immunohistochemical staining combined with tissue microarray of lung cancer, Terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL) was applied to detect the apoptosis in the tissue microarray, The apoptosis index(AI) were defined. The methylations of ASPPl and ASPP2 promoter region CpG island were analysised by methylation-specific PCR (MSP). Furthermore, We analysis the relationship among expression of ASPP1、ASPP2 protein, promoter region methylation,AI,and p53 apoptosis pathway.
Results: 1.The position rate of ASPP1 in 90 cases lung cancer tissues was 27.8%, and the position rate of ASPP2 was 11.1%; the positive rate of ASPP1 in 25 normal adjacent lung tissues was 52.0%, and the ASPP2 positive rate was 16.0%; Significant difference was found between the expression of ASPP1 in cancer tissue and their normal adjacent tissues (x2=5.188,P<0.05); while the expression of ASPP2 has no significant difference (x2=0.437,P>0.05).
2. The expression of ASPP1 in 90 cases lung cancer had positive relation with the differentiation, TNM stage and lymph node metastasis of the disease (x2:=4.680、6.330、6.045, P<0.05),but not related to sex, age,and histological type. Significant difference was not found between the expression of ASPP2 and clinical pathologic characteristic(P>0.05).
3. The positive expression of ASPP1 was 16 cases in 39 cases that negative expression of p53,and 9 cases in 51cases that mutation p53 cancer tissues. The Spearman's method was used, and the result was that there was negative correlation between the expression of ASPP1 and mutation p53 in lung cancer (r=-0.259,P<0.01),but the positive expression of ASPP2 was 6 cases in 39 cases that nagetive expression of p53 cancer tissues, and 4 cases in 51 cases mutation p53 tissues, the expression of ASPP2 was not associated with p53 state by used Spearman's method(r=-0.119,P>0.05).
4. The AI in the group of ASPP1 positive expression was significantly higher than in the group of ASPP 1 negative expression, this showed that there was close relationship between the expression of ASPP1 and cell apoptosis in lung carcinoma (t=2.337,P<0.05); but no significant different was found between the group of ASPP2 positive expression and the group of negative expression, so the expression of ASPP2 may has no relation with cell apoptosis in lung carcinoma(t=1.083,P>0.05)
5. The positive rate of ASPP 1 methylation in lung tumor tissues was significantly higher than that in the adjacent lung tissues{42.22%(38/90) vs 16.00%(4/25), P =0.019}.The methylation state of ASPP 1 had close relation with TNM stage and lymphnode metastasis of the disease (x2=6.919、5.213,P=<0.05),but not related to age,sex, histological type and differentiation(x2=1.l29、1.488、1.126、4.555,P >0.05).The group with promoter methylation showed lower expression of ASPP 1 than that with unmethylation state (x2=5.188,P<0.05); Methylation in the promoter region of ASPP2 gene was not detected in lung cancer (n=90) or side-carcinoma tissues (n=25);
Conclutions: 1. The lower expression of ASPP1 was correlation with TNM stage and lymph node metastasis in lung cancer, that shows the loss expression of ASPP1 may participate in lung carcinogenesis and development.
2. There was no significant difference in expression of ASPP2 between lung cancer tissue and the paracarcinomatous tissues, it indicated that ASPP2 may not be as a tumor suppressor gene in lung carcinomas.
3. ASPP1 can induce AI, and the expression of ASPP1 has a negative correlationship with mutation p53; ASPP1 may be as a key factor of p53 apoptosis pathway in lung cancer tissue;but ASPP2 may be independent of the p53 apoptosis pathway.
4. The hypermethylation of ASPP1 gene might lead to the abnormal expression of ASPP1 protein; and it is respected of the aggressive clinical behavior and advanced tumor stage; while the ASPP2 gene promoter methylation may not be the cause of the abnormal expression of ASPP2 protein in tumorgenesis.
方法：本研究实验组收集肺癌组织90例,对照组25例相应癌旁正常肺组织,构建肺癌组织芯片,免疫组织化学染色Envision法检测ASPP1、ASPP2及p53蛋白在肺癌组织芯片中的表达情况,并同时利用原位末端转移酶标记检测法(Terminal deoxynucleotidyl transferase dUTP nick end labeling,TUNEL法)检测肺癌细胞的凋亡指数AI,应用甲基化特异性PCR(methylation-specific PCR,MSP)对抑癌基因ASPP1、ASPP2启动子CpG岛进行甲基化状态检测,分析ASPP1、ASPP2蛋白表达、启动子甲基化和p53细胞凋亡通路之间的关系。
结果
1.90例肺癌组织ASPP1和ASPP2阳性率分别为27.8％和11.1％；25例癌旁组织中ASPP1和ASPP2阳性率分别为52.0％和16.0％；ASPP1在肺癌组织和癌旁组织中表达阳性率两者比较,差异有统计学意义(χ2=5.188,P<0.05),而ASPP2在肺癌组织和癌旁组织中表达阳性率两者比较,差异无统计学意义(χ2=0.437,P>0.05)。
2.90例肺癌组织中ASPP1蛋白的表达和分化程度、TNM分期及淋巴结转移呈正相关(P<0.05),与患者的性别、年龄、肿瘤大小、组织学类型无关(P>0.05)；而ASPP2蛋白的表达与肺癌的临床病理特征均无关(P>0.05)。
3.39例p53表达阴性的肺癌组织中ASPP1表达阳性16例(41.02％),51例突变型p53肺癌组织中ASPP1表达阳性9例(17.64％),经Spearman's相关分析,说明在肺癌组织中ASPP1蛋白表达与突变型p53蛋白表达呈负相关(r=-0.259,P<0.01),而39例p53表达阴性的肺癌患者中ASPP2蛋白表达阳性6例(15.38％),51例突变型p53中4例ASPP2表达阳性(9.75％),经Spearman's相关分析,可认为在肺癌组织中ASPP2蛋白表达与p53蛋白的表达无相关性(r=-0.119,P>0.05)。
4.比较90肺癌组织ASPP1蛋白阳性表达组与阴性组中AI值,发现ASPP1蛋白阳性组高于阴性组,提示ASPP1蛋白表达与细胞凋亡有关(t=2.337,P<0.05)；而90例肺癌组织中,ASPP2蛋白表达阳性组和阴性组之间AI值无显著性差异(t=1.083,P>0.05),可以为ASPP2蛋白的表达与肺癌细胞凋亡无关。
5.NSCLC中ASPP1基因启动子甲基化率为42.22％(38／90),癌旁组织甲基化率为16.00％(4／25),有显著性差异(χ2=5.830,P>0.05)；ASPP1基因启动子甲基化与患者的年龄、性别、组织类型和分化程度无关(χ2=1.129、1.488、1.126、4.555,P>0.05),但与患者的TNM分期和淋巴结转移有关(χ2=6.919、5.213,P=<0.05)；90例肺癌组织中ASPP1甲基化者其蛋白表达率明显低于未甲基化者(χ2=5.188,P<0.05)；而ASPP2基因启动子在90例NSCLC和25例癌旁组织中均未检测到甲基化状态。
结论
1.ASPP1蛋白在肺癌组织中低表达,与肿瘤分化程度、临床分期和淋巴结转移有关,提示ASPP1表达缺失可能与肺癌发生、进展有关。
2.ASPP2蛋白在肺癌组织和癌旁肺组织中的表达无明显差异,提示ASPP2在肺癌中可能不具有肿瘤抑制基因特性。
3.在肺癌组织中,ASPP1促进肺癌细胞AI的增高,且其表达与突变型p53呈负相关,ASPP1可能是p53凋亡通路中的关键蛋白；而ASPP2可能与p53凋亡通路无关。
4.肺癌组织中ASPP1启动子甲基化可能是ASPP1蛋白表达下调的原因,ASPP1高甲基化预示着NSCLC的高侵袭性,而ASPP2基因甲基化可能不是其蛋白的表达的调控机制。
Objective:In order to research the relation between ASPP gene family and incidence of lung carcinoma, the article examined the promoter methylation statut of the ASPP1 and ASPP2 from lever of DNA in lung carcinoma, and the expression of ASPP1、ASPP2 and p53 from lever of protein, and the apoptosis index of the lung cancer tissues.
Metholds:To construct tissue microarray of lung cancers,we collected 90 cases lung carcinoma tissues as experimental group and 25 normal adjacent lung tissue as control group. The expression state of ASPP1、ASPP2 and p53 protion were determined by immunohistochemical staining combined with tissue microarray of lung cancer, Terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL) was applied to detect the apoptosis in the tissue microarray, The apoptosis index(AI) were defined. The methylations of ASPPl and ASPP2 promoter region CpG island were analysised by methylation-specific PCR (MSP). Furthermore, We analysis the relationship among expression of ASPP1、ASPP2 protein, promoter region methylation,AI,and p53 apoptosis pathway.
Results: 1.The position rate of ASPP1 in 90 cases lung cancer tissues was 27.8%, and the position rate of ASPP2 was 11.1%; the positive rate of ASPP1 in 25 normal adjacent lung tissues was 52.0%, and the ASPP2 positive rate was 16.0%; Significant difference was found between the expression of ASPP1 in cancer tissue and their normal adjacent tissues (x2=5.188,P<0.05); while the expression of ASPP2 has no significant difference (x2=0.437,P>0.05).
2. The expression of ASPP1 in 90 cases lung cancer had positive relation with the differentiation, TNM stage and lymph node metastasis of the disease (x2:=4.680、6.330、6.045, P<0.05),but not related to sex, age,and histological type. Significant difference was not found between the expression of ASPP2 and clinical pathologic characteristic(P>0.05).
3. The positive expression of ASPP1 was 16 cases in 39 cases that negative expression of p53,and 9 cases in 51cases that mutation p53 cancer tissues. The Spearman's method was used, and the result was that there was negative correlation between the expression of ASPP1 and mutation p53 in lung cancer (r=-0.259,P<0.01),but the positive expression of ASPP2 was 6 cases in 39 cases that nagetive expression of p53 cancer tissues, and 4 cases in 51 cases mutation p53 tissues, the expression of ASPP2 was not associated with p53 state by used Spearman's method(r=-0.119,P>0.05).
4. The AI in the group of ASPP1 positive expression was significantly higher than in the group of ASPP 1 negative expression, this showed that there was close relationship between the expression of ASPP1 and cell apoptosis in lung carcinoma (t=2.337,P<0.05); but no significant different was found between the group of ASPP2 positive expression and the group of negative expression, so the expression of ASPP2 may has no relation with cell apoptosis in lung carcinoma(t=1.083,P>0.05)
5. The positive rate of ASPP 1 methylation in lung tumor tissues was significantly higher than that in the adjacent lung tissues{42.22%(38/90) vs 16.00%(4/25), P =0.019}.The methylation state of ASPP 1 had close relation with TNM stage and lymphnode metastasis of the disease (x2=6.919、5.213,P=<0.05),but not related to age,sex, histological type and differentiation(x2=1.l29、1.488、1.126、4.555,P >0.05).The group with promoter methylation showed lower expression of ASPP 1 than that with unmethylation state (x2=5.188,P<0.05); Methylation in the promoter region of ASPP2 gene was not detected in lung cancer (n=90) or side-carcinoma tissues (n=25);
Conclutions: 1. The lower expression of ASPP1 was correlation with TNM stage and lymph node metastasis in lung cancer, that shows the loss expression of ASPP1 may participate in lung carcinogenesis and development.
2. There was no significant difference in expression of ASPP2 between lung cancer tissue and the paracarcinomatous tissues, it indicated that ASPP2 may not be as a tumor suppressor gene in lung carcinomas.
3. ASPP1 can induce AI, and the expression of ASPP1 has a negative correlationship with mutation p53; ASPP1 may be as a key factor of p53 apoptosis pathway in lung cancer tissue;but ASPP2 may be independent of the p53 apoptosis pathway.
4. The hypermethylation of ASPP1 gene might lead to the abnormal expression of ASPP1 protein; and it is respected of the aggressive clinical behavior and advanced tumor stage; while the ASPP2 gene promoter methylation may not be the cause of the abnormal expression of ASPP2 protein in tumorgenesis.
引文
[1]Jonat han C, Daniel W, Roy S. Gene Therapy of Lung Cancer.Semin Surg Oncol,2003,21 (3):1962204
[2]Shivapurkar N,Reddy J,Chaudhary P M, et al.Apoptosis and lung cancer[J].J Cell Biochem,2003,88(5):885-898.
[3]Esteller M,Fraga MF,Guo M,et al.DNA methylation patterns in hereditary human cancers mimic sporadic tumorigenesis.Hum Mol Genet,2001,10(26):3001-3007
[4]Mitsudomi T, Hamajima N, Ogava M, et al. Prognostic significance of p53 alterations in patients with non-small cell lung cancer:a metaanalysis [J]. Clin Cancer Res,2000,6: 4055-4063
[5]Samuels-Lev Y, O'Connor DJ, Bergamaschi D, et al. ASPP proteins specifically atimulate the apoptotic function of p53[J]. Mol Cell,2001,8(4):781-794
[6]Robertson KD,W. A.,DNA methylation in health and disease.Nat Rev Genet,2000,1:11-19.
[7]Robertson KD,J. P.,DNA methylation:past, present, and future diredtions. Carcinogenesis, 2000,21:461-467
[8]Roloff TC,R. H.,Nuber UA.Compartibe study of methyl-CpG-binding domain proteins.BMC Genomics,2003,4:1-7
[9]Palmisano WA,D.K.,Szccomanno G,et al.Predicting lung cancer by delecting aberrant promoter methylation in sputum.Cancer Res,2000,60:5954-5958.
[10]Esteller M.Epigenetic gene sillencing in cancer:the DNA hypermethylome[J].Hum Mol.Genet,2007,16:50-59
[11]Herman JG,Graff JR,Myohanen S,et al.Methylation specific PCR:anovel PCR assay for methylation status of CpG islands[J].Proc Natl Acad Sci USA,1996,93(18):9821-9826
[12]Agirre X, Roman-Gomez J, Jimenez-Velasco A, et al. ASPP1,a common activator of TP53, in inactivated by aberrant methylation of its promoter in acute lymphoblastic leukemia[J]. Oncogene,2006,25(13):1862-1870
[13]Konanen J,Bubendorf L,Kallioniemi A,et al.Tissue microarrays for high-throughput molecular profiling of tumor specimens[J]. Natl Med,1998,4:844-847.
[14]Fontanini QBlodrini L,Calcinal A,et al.Thrombospondins Ⅰand Ⅱmessagenger RNA express in lung cancinoma:relationship with p53 alterations,angiogenic growth factors,and vascular density[J].Clin Cancer Res,1999,5(1):155-161
[15]Bergamaschi D, Samuels Y, O'Neil NJ, et al. iASPP oncoprotain is a key inhibitor of p53 conserved from worm to human[J]. Nat Genet,2003,33(2):162-167
[16]Tidow H, Andreeva A, Rutheeford TJ, et al.Solution structure of ASPP2 N-terminal domain (N-ASPP2) reveals a ubiquitin-like fold[J]. J Mol Biol,2007,371(4):948-958.
[17]Slee EA, Gillotin S, Bergamaschi D, et al. The N-terminus of isoform of human iASPP is required for its cytoplasmic localization[J]. Oncogene,2004,23 (56):9007-9016
[18]Slee EA,Lu X. The ASPP family:deciding between life and death after DNA damage.Toxicol Lett,2003,139(2-3):81-89
[19]Mori S, Ito G, Usanmi N, et al. p53 apoptotic pathway molecules are frequently and simultaneously altered in nonsmall cell lung carcinoma [J]. Cancer,2004,100(8):1673-82
[20]Mori T,Okamoto H, Takahashi N, et al. Aberrant overexpression of 53BP2 mRNA in lung cancer cell lines [J]. FEBS Lett,2000,465(2-3):124-128
[21]Virginie Vives,Elizabeth Slee,Xin Lu,ASPP2 gene that control life and death in vivo[J].Cell Cycle,2006,5(19):2187-2190
[22]Santarosa M,Ashworth A. Haploinsufficiency for tumor supperessor genes:when you don't need to go all the way[J].Biochim Biophys Acta,2004,1654:105-22
[23]Chen X,Ko LJ,Jayaraman L,Prives C,et al.p53 levels,functional domains,and DNA damage determine the extent of the apoptotic response of tumor cells [J]. Genes Dev,1996,10(19):2438-2451
[24]Ludwig RL,Bates S,Vousden,KH,et al.Differential activation of target cellular promoters by p53 mutants with impaired apoptotic function[J].Mol Cell Biol.1996,16 (9):4952-4962
[25]Vojtesek B,Lane DP.Regulation of p53 protein expression in human breast cancer cell lines[J]. J Cell Sci,1993,105:107
[26]Merol GR.p53-dependent and p53-independent activation of apoptosis in mamary epithelial cell reveals a survival function of EPG and insulin[J].J Cell Biel,1995,128(6):1185
[27]Smart P,Lane EB,Lane DP,et al. Effects on normal fibroblasts and neuroblastoma cells of the activation of the p53 response by the nuclear export inhibitor leptomycin B.Oncogene.1999,18(15):7378-7386
[28]Saller E,Tom E,Brunori M,et al.Increased apoptosis induction by 121F mutant p53[J].EMBO J.1999,18(16):4424-4437
[29]Caelles C, Helmberg A, Karin M.p53-dependent apoptosis in the absence of transcriptional activation of p53-target genes[J].Nature.1994,370(6486):220-3.
[30]Iwabuchi K, Li B, Massa H F, et al. Stimulation of p53-mediated transcriptional activation by the p53-binding proteins,53BP1 and 52BP2[J].J Biol Chem,1998,273(40):26061-26068.
[31]苏胜发,卢冰,苏敏,等.非小细胞肺癌中ASPP1、ASPP2蛋白的不同表达状态及其意义[J].肿瘤预防与治疗,2008,21(2)：216-219
[32]Lopez CD,Ao Y,Rohde LH,et al.Proapoptotic p53 interacting protein 53BP2 is induced by UV irradiation but suppressed by p53.Mol Cell Biol 2000,20(21):8018-8025
[33]Staunton MJ,Gaffneye F. Tumor type is a determinant of susceotibility to apoptosis[J]. Am J Clin Pathol,1995,103:300-303
[34]Dworakowska D,Jassem E,Jassem J et al. prognostic value of the apoptotic index analysed jointly with selected cell cycle regulators and proliferation markers in non-small cell lung cancer[J]. Lung cancer,2009 Feb 4,[Epub abead of print]
[35]Johnson J,Lagowski J,Lawson S,et al.p73 expression modulates p63 and Mdm2 protein presence in complex with family specific DNA target sequence in squamous cell carcinogenesis[J].Oncogene,2008,27(19):2780-7
[36]Delgado BM,Piueras JF,Carcia MJ,et al.Hypermethylation of 5'CpG island of P16 is a frequent event in non-Hodgkin's lymphoma. Lekemia,1997,11(3):425-42
[37]Herman JG, Baylin SB.Gene silencing in cancer in association with promoter hypermethylation[J].N Eng J Med,2003,349(21):2042
[38]Liu ZJ, Lu X, Zhang Y, et al. Downregalated mRNA expression of ASPP and the hypermethylation of the 5'-untranslated region in cancer cell lines retaining wild-type p53[J]. FEBS Lett,2005,579(7):1587-1590
[39]Zochbauer-Muller S,Fong KM,VimaniAK,et al.Aberrant ptomoter methylation of multiple genes in non-small cell lung cancer [J]. Cancer Res,2001,61(1)249
[40]Fogal V, Kartasheva NN, Trigiante G, et al. ASPP1 and ASPP2 are new transcriptional targets
of E2F [J]. Cell Death Differ,2005 Apr,12(4):369-376
[41]Hershko T,Chaussepied M,Oren M,et al.Ginsberg D Novel link between E2F and p53: proapoptotic cofactors of p53 are transcriptionally upregulated by E2F.Cell Death Differ.2005,12:377-383
[42]Baylin SB, Herman JG, Graff JR, Vertino PM, Issa JP:Aleration in DNA methylation:a fundamental aspect of neoplasia[J], Adv Cancer Res 1998,72(1):141-196
[43]Wolffe AP,Matzke MA.Epigenetices regulation through repression [J]. Science,1999, 286(5439):481-486
[44]Jones PA. The role of DNA methylation in mammalian epigentics [J].Science, 2001;293(5532):1068-1070.
[45]Gonzalgo ML,Hayashiba T, Bender CM.The role of DNA methylation in exprssion of P19/P16 locus in human bladder cancer cell line.Caner Res 1998;58:1245-1252.
[46]Bachman KE,Park BH,Rhee I,et al.Histone modification and silencing prior to DNA methylation of a tumor suppressor gene[J].Cancer Cell,2003,3:89-95
[47]Roman-Gomez J,Jimenez-velasco A,Agirre X,et al.Lack of CpG island methylator phenotype defines a clinical subtype of T-cell acute lymphoblastic leukemia associated with good prognosis[J].J Clin Oncol,2005,3(28):7043-7049
[48]邬国斌.肝细胞癌中ASPP1、ASPP2基因表达及启动子区CpG岛甲基化的实验研究[D].中国博士学位论文数据库,2008,(10)
[49]Tidow H, Andreeva A, Rutheeford TJ, et al. Solution structure of ASPP2 N-terminal domain (N-ASPP2) reveals a ubiquitin-like fold[J]. J Mol Biol,2007,371(4):948-958
[1]Samuels-Lev Y, O'Connor DJ, Bergamaschi D, et al. ASPP proteins specifically atimulate the apoptotic function of p53[J]. Mol Cell,2001,8(4):781-794
[2]Bergamaschi D, Samuels Y, O'Neil NJ, et al. iASPP oncoprotain is a key inhibitor of p53 conserved from worm to human[J]. Nat Genet,2003,33(2):162-167
[3]Nao Ko, Takahashi, Shinya Kobayashi, et al. Expression of 53BP2 and ASPP2 proteins from Tp53BP2 gene by alternative splicing[J]. Biochem Biophys Res Commun, 2004(2),315:434-438
[4]Tidow H, Andreeva A, Rutheeford TJ, et al. Solution structure of ASPP2 N-terminal domain (N-ASPP2) reveals a ubiquitin-like fold[J]. J Mol Biol,2007,371(4):948-958.
[5]Slee EA, Gillotin S, Bergamaschi D, et al. The N-terminus of isoform of human iASPP is required for its cytoplasmic localization[J]. Oncogene,2004,23(56):9007-9016
[6]赵劲风综述,张阳德审校.肝癌组织中p53基因及其下游基因的研究进展[J].中国现代医学杂志,2003,13(24)：70-73
[7]Gorina S, Pavetich N P. Structure of the p53 tumor suppressor bound to the ankyin and SH3 domains of 53BP2[J]. Science,1996,274(5289):1001-1005
[8]Robinson RA, Lu X, Jones EY, et al. Biochemical and structural studies of ASPP proteins reveal differential binding to p53,p63,p73[J]. Structure,2008,16(2):259-68
[9]Bergamaschi D, Samuels Y, Zhong S, et al. Mdm2 and mdmx prevent ASPP 1 And ASPP2 from stimulating p53 without targeting p53 for degradation [J]. Oncogene,2005,24(23):3836-42
[10]Takahashi N,Kobayashi S,Kajino S,Imai K et al Inhibition of the 53BP2S-mediated apoptosis by nuclear factor kappaB and family proteins[J]. Genes Cells,2005,10:803-811
[11]A Sullivan, X Lu. ASPP:a new family of oncogenes and tumor suppressor genes [J]. Br J Cancer,2007,96(2):196-200
[12]Katz C, Benyamini H, Rotem S, et al. Molecular basis of the interaction between the antiapoptotic Bcl-2 family proteins and the proapoptotic protein ASPP2[J]. Structure,2008 Feb,16(2):259-268
[13]Fogal V, Kartasheva NN, Trigiante G, et al. ASPP1 and ASPP2 are new transcriptional targets of E2F[J]. Cell Death Differ,2005,12(4):369-376
[14]Liu ZJ, Lu X, Zhang Y, et al. Downregalated mRNA expression of ASPP and the hypermethylation of the 5'-untranslated region in cancer cell lines retaining wild-type p53[J]. FEBS Lett,2005,579(7):1587-1590
[15]Agirre X, Roman-Gomez J, Jimenez-Velasco A, et al. ASPP1,a common activator of TP53, in inactivated by aberrant methylation of its promoter in acute lymphoblastic leukemia[J]. Oncogene,2006,25(13):1862-1870
[16]Braithwaite AW, Del Sal G, Lu X. Some p53-binding proteins than can function as arbiters of life and death[J]. Cell Death and Differentiation,2006,13(6):984-993
[17]Zhu Z, Ramos J, Kampa K, et al. Control of ASPP2/(53BP2L) protein levels by proteasomal degradation modulates p53 apoptotic function [J]. J Biol Chem,2005,280(41):34473-34480
[18]Liu ZJ, Zhang Y, Zhang XB, et al. Abnormal mRNA expression of ASPP member in leukemia cell lines[J]. Leukemia,2004,18(4):880
[19]Mori S, Ito G, Usanmi N, et al. p53 apoptotic pathway molecules are frequently and simultaneously altered in nonsmall cell lung carcinoma [J]. Cancer,2004,100(8):1673-82
[20]Mori T, Okamoto H, Takahashi N, et al. Aberrant overexpression of 53BP2 mRNA in lung cancer cell lines [J]. FEBS Lett,2000,465(2-3):124-128
[21]Ju H, Lee KA, Yang M, et al. TP53BP2 locus is associated with gastric cancer susceptibility [J]. Int J Cancer,2005,117(6):957-960
[22]Liu ZJ,Cai Y,Hou L, et al. Effect of RNA interferrence of iASPP on the apoptosis in MCF-7 breast cancer cells[J]. Cancer Invest,2008,26(9):87
[2]Shivapurkar N,Reddy J,Chaudhary P M, et al.Apoptosis and lung cancer[J].J Cell Biochem,2003,88(5):885-898.
[3]Esteller M,Fraga MF,Guo M,et al.DNA methylation patterns in hereditary human cancers mimic sporadic tumorigenesis.Hum Mol Genet,2001,10(26):3001-3007
[4]Mitsudomi T, Hamajima N, Ogava M, et al. Prognostic significance of p53 alterations in patients with non-small cell lung cancer:a metaanalysis [J]. Clin Cancer Res,2000,6: 4055-4063
[5]Samuels-Lev Y, O'Connor DJ, Bergamaschi D, et al. ASPP proteins specifically atimulate the apoptotic function of p53[J]. Mol Cell,2001,8(4):781-794
[6]Robertson KD,W. A.,DNA methylation in health and disease.Nat Rev Genet,2000,1:11-19.
[7]Robertson KD,J. P.,DNA methylation:past, present, and future diredtions. Carcinogenesis, 2000,21:461-467
[8]Roloff TC,R. H.,Nuber UA.Compartibe study of methyl-CpG-binding domain proteins.BMC Genomics,2003,4:1-7
[9]Palmisano WA,D.K.,Szccomanno G,et al.Predicting lung cancer by delecting aberrant promoter methylation in sputum.Cancer Res,2000,60:5954-5958.
[10]Esteller M.Epigenetic gene sillencing in cancer:the DNA hypermethylome[J].Hum Mol.Genet,2007,16:50-59
[11]Herman JG,Graff JR,Myohanen S,et al.Methylation specific PCR:anovel PCR assay for methylation status of CpG islands[J].Proc Natl Acad Sci USA,1996,93(18):9821-9826
[12]Agirre X, Roman-Gomez J, Jimenez-Velasco A, et al. ASPP1,a common activator of TP53, in inactivated by aberrant methylation of its promoter in acute lymphoblastic leukemia[J]. Oncogene,2006,25(13):1862-1870
[13]Konanen J,Bubendorf L,Kallioniemi A,et al.Tissue microarrays for high-throughput molecular profiling of tumor specimens[J]. Natl Med,1998,4:844-847.
[14]Fontanini QBlodrini L,Calcinal A,et al.Thrombospondins Ⅰand Ⅱmessagenger RNA express in lung cancinoma:relationship with p53 alterations,angiogenic growth factors,and vascular density[J].Clin Cancer Res,1999,5(1):155-161
[15]Bergamaschi D, Samuels Y, O'Neil NJ, et al. iASPP oncoprotain is a key inhibitor of p53 conserved from worm to human[J]. Nat Genet,2003,33(2):162-167
[16]Tidow H, Andreeva A, Rutheeford TJ, et al.Solution structure of ASPP2 N-terminal domain (N-ASPP2) reveals a ubiquitin-like fold[J]. J Mol Biol,2007,371(4):948-958.
[17]Slee EA, Gillotin S, Bergamaschi D, et al. The N-terminus of isoform of human iASPP is required for its cytoplasmic localization[J]. Oncogene,2004,23 (56):9007-9016
[18]Slee EA,Lu X. The ASPP family:deciding between life and death after DNA damage.Toxicol Lett,2003,139(2-3):81-89
[19]Mori S, Ito G, Usanmi N, et al. p53 apoptotic pathway molecules are frequently and simultaneously altered in nonsmall cell lung carcinoma [J]. Cancer,2004,100(8):1673-82
[20]Mori T,Okamoto H, Takahashi N, et al. Aberrant overexpression of 53BP2 mRNA in lung cancer cell lines [J]. FEBS Lett,2000,465(2-3):124-128
[21]Virginie Vives,Elizabeth Slee,Xin Lu,ASPP2 gene that control life and death in vivo[J].Cell Cycle,2006,5(19):2187-2190
[22]Santarosa M,Ashworth A. Haploinsufficiency for tumor supperessor genes:when you don't need to go all the way[J].Biochim Biophys Acta,2004,1654:105-22
[23]Chen X,Ko LJ,Jayaraman L,Prives C,et al.p53 levels,functional domains,and DNA damage determine the extent of the apoptotic response of tumor cells [J]. Genes Dev,1996,10(19):2438-2451
[24]Ludwig RL,Bates S,Vousden,KH,et al.Differential activation of target cellular promoters by p53 mutants with impaired apoptotic function[J].Mol Cell Biol.1996,16 (9):4952-4962
[25]Vojtesek B,Lane DP.Regulation of p53 protein expression in human breast cancer cell lines[J]. J Cell Sci,1993,105:107
[26]Merol GR.p53-dependent and p53-independent activation of apoptosis in mamary epithelial cell reveals a survival function of EPG and insulin[J].J Cell Biel,1995,128(6):1185
[27]Smart P,Lane EB,Lane DP,et al. Effects on normal fibroblasts and neuroblastoma cells of the activation of the p53 response by the nuclear export inhibitor leptomycin B.Oncogene.1999,18(15):7378-7386
[28]Saller E,Tom E,Brunori M,et al.Increased apoptosis induction by 121F mutant p53[J].EMBO J.1999,18(16):4424-4437
[29]Caelles C, Helmberg A, Karin M.p53-dependent apoptosis in the absence of transcriptional activation of p53-target genes[J].Nature.1994,370(6486):220-3.
[30]Iwabuchi K, Li B, Massa H F, et al. Stimulation of p53-mediated transcriptional activation by the p53-binding proteins,53BP1 and 52BP2[J].J Biol Chem,1998,273(40):26061-26068.
[31]苏胜发,卢冰,苏敏,等.非小细胞肺癌中ASPP1、ASPP2蛋白的不同表达状态及其意义[J].肿瘤预防与治疗,2008,21(2)：216-219
[32]Lopez CD,Ao Y,Rohde LH,et al.Proapoptotic p53 interacting protein 53BP2 is induced by UV irradiation but suppressed by p53.Mol Cell Biol 2000,20(21):8018-8025
[33]Staunton MJ,Gaffneye F. Tumor type is a determinant of susceotibility to apoptosis[J]. Am J Clin Pathol,1995,103:300-303
[34]Dworakowska D,Jassem E,Jassem J et al. prognostic value of the apoptotic index analysed jointly with selected cell cycle regulators and proliferation markers in non-small cell lung cancer[J]. Lung cancer,2009 Feb 4,[Epub abead of print]
[35]Johnson J,Lagowski J,Lawson S,et al.p73 expression modulates p63 and Mdm2 protein presence in complex with family specific DNA target sequence in squamous cell carcinogenesis[J].Oncogene,2008,27(19):2780-7
[36]Delgado BM,Piueras JF,Carcia MJ,et al.Hypermethylation of 5'CpG island of P16 is a frequent event in non-Hodgkin's lymphoma. Lekemia,1997,11(3):425-42
[37]Herman JG, Baylin SB.Gene silencing in cancer in association with promoter hypermethylation[J].N Eng J Med,2003,349(21):2042
[38]Liu ZJ, Lu X, Zhang Y, et al. Downregalated mRNA expression of ASPP and the hypermethylation of the 5'-untranslated region in cancer cell lines retaining wild-type p53[J]. FEBS Lett,2005,579(7):1587-1590
[39]Zochbauer-Muller S,Fong KM,VimaniAK,et al.Aberrant ptomoter methylation of multiple genes in non-small cell lung cancer [J]. Cancer Res,2001,61(1)249
[40]Fogal V, Kartasheva NN, Trigiante G, et al. ASPP1 and ASPP2 are new transcriptional targets
of E2F [J]. Cell Death Differ,2005 Apr,12(4):369-376
[41]Hershko T,Chaussepied M,Oren M,et al.Ginsberg D Novel link between E2F and p53: proapoptotic cofactors of p53 are transcriptionally upregulated by E2F.Cell Death Differ.2005,12:377-383
[42]Baylin SB, Herman JG, Graff JR, Vertino PM, Issa JP:Aleration in DNA methylation:a fundamental aspect of neoplasia[J], Adv Cancer Res 1998,72(1):141-196
[43]Wolffe AP,Matzke MA.Epigenetices regulation through repression [J]. Science,1999, 286(5439):481-486
[44]Jones PA. The role of DNA methylation in mammalian epigentics [J].Science, 2001;293(5532):1068-1070.
[45]Gonzalgo ML,Hayashiba T, Bender CM.The role of DNA methylation in exprssion of P19/P16 locus in human bladder cancer cell line.Caner Res 1998;58:1245-1252.
[46]Bachman KE,Park BH,Rhee I,et al.Histone modification and silencing prior to DNA methylation of a tumor suppressor gene[J].Cancer Cell,2003,3:89-95
[47]Roman-Gomez J,Jimenez-velasco A,Agirre X,et al.Lack of CpG island methylator phenotype defines a clinical subtype of T-cell acute lymphoblastic leukemia associated with good prognosis[J].J Clin Oncol,2005,3(28):7043-7049
[48]邬国斌.肝细胞癌中ASPP1、ASPP2基因表达及启动子区CpG岛甲基化的实验研究[D].中国博士学位论文数据库,2008,(10)
[49]Tidow H, Andreeva A, Rutheeford TJ, et al. Solution structure of ASPP2 N-terminal domain (N-ASPP2) reveals a ubiquitin-like fold[J]. J Mol Biol,2007,371(4):948-958
[1]Samuels-Lev Y, O'Connor DJ, Bergamaschi D, et al. ASPP proteins specifically atimulate the apoptotic function of p53[J]. Mol Cell,2001,8(4):781-794
[2]Bergamaschi D, Samuels Y, O'Neil NJ, et al. iASPP oncoprotain is a key inhibitor of p53 conserved from worm to human[J]. Nat Genet,2003,33(2):162-167
[3]Nao Ko, Takahashi, Shinya Kobayashi, et al. Expression of 53BP2 and ASPP2 proteins from Tp53BP2 gene by alternative splicing[J]. Biochem Biophys Res Commun, 2004(2),315:434-438
[4]Tidow H, Andreeva A, Rutheeford TJ, et al. Solution structure of ASPP2 N-terminal domain (N-ASPP2) reveals a ubiquitin-like fold[J]. J Mol Biol,2007,371(4):948-958.
[5]Slee EA, Gillotin S, Bergamaschi D, et al. The N-terminus of isoform of human iASPP is required for its cytoplasmic localization[J]. Oncogene,2004,23(56):9007-9016
[6]赵劲风综述,张阳德审校.肝癌组织中p53基因及其下游基因的研究进展[J].中国现代医学杂志,2003,13(24)：70-73
[7]Gorina S, Pavetich N P. Structure of the p53 tumor suppressor bound to the ankyin and SH3 domains of 53BP2[J]. Science,1996,274(5289):1001-1005
[8]Robinson RA, Lu X, Jones EY, et al. Biochemical and structural studies of ASPP proteins reveal differential binding to p53,p63,p73[J]. Structure,2008,16(2):259-68
[9]Bergamaschi D, Samuels Y, Zhong S, et al. Mdm2 and mdmx prevent ASPP 1 And ASPP2 from stimulating p53 without targeting p53 for degradation [J]. Oncogene,2005,24(23):3836-42
[10]Takahashi N,Kobayashi S,Kajino S,Imai K et al Inhibition of the 53BP2S-mediated apoptosis by nuclear factor kappaB and family proteins[J]. Genes Cells,2005,10:803-811
[11]A Sullivan, X Lu. ASPP:a new family of oncogenes and tumor suppressor genes [J]. Br J Cancer,2007,96(2):196-200
[12]Katz C, Benyamini H, Rotem S, et al. Molecular basis of the interaction between the antiapoptotic Bcl-2 family proteins and the proapoptotic protein ASPP2[J]. Structure,2008 Feb,16(2):259-268
[13]Fogal V, Kartasheva NN, Trigiante G, et al. ASPP1 and ASPP2 are new transcriptional targets of E2F[J]. Cell Death Differ,2005,12(4):369-376
[14]Liu ZJ, Lu X, Zhang Y, et al. Downregalated mRNA expression of ASPP and the hypermethylation of the 5'-untranslated region in cancer cell lines retaining wild-type p53[J]. FEBS Lett,2005,579(7):1587-1590
[15]Agirre X, Roman-Gomez J, Jimenez-Velasco A, et al. ASPP1,a common activator of TP53, in inactivated by aberrant methylation of its promoter in acute lymphoblastic leukemia[J]. Oncogene,2006,25(13):1862-1870
[16]Braithwaite AW, Del Sal G, Lu X. Some p53-binding proteins than can function as arbiters of life and death[J]. Cell Death and Differentiation,2006,13(6):984-993
[17]Zhu Z, Ramos J, Kampa K, et al. Control of ASPP2/(53BP2L) protein levels by proteasomal degradation modulates p53 apoptotic function [J]. J Biol Chem,2005,280(41):34473-34480
[18]Liu ZJ, Zhang Y, Zhang XB, et al. Abnormal mRNA expression of ASPP member in leukemia cell lines[J]. Leukemia,2004,18(4):880
[19]Mori S, Ito G, Usanmi N, et al. p53 apoptotic pathway molecules are frequently and simultaneously altered in nonsmall cell lung carcinoma [J]. Cancer,2004,100(8):1673-82
[20]Mori T, Okamoto H, Takahashi N, et al. Aberrant overexpression of 53BP2 mRNA in lung cancer cell lines [J]. FEBS Lett,2000,465(2-3):124-128
[21]Ju H, Lee KA, Yang M, et al. TP53BP2 locus is associated with gastric cancer susceptibility [J]. Int J Cancer,2005,117(6):957-960
[22]Liu ZJ,Cai Y,Hou L, et al. Effect of RNA interferrence of iASPP on the apoptosis in MCF-7 breast cancer cells[J]. Cancer Invest,2008,26(9):87