散发性乳腺癌中DNMT3a、3b表达及其与BRCA1基因启动子甲基化和蛋白表达的相关性
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摘要
目的:探讨DNMT3a、3b蛋白表达与散发性乳腺癌BRCA1基因启动子甲基化状态及蛋白表达的相关性。方法:收集乳腺肿瘤组织及相关临床资料。采用免疫组化法检测200例散发性乳腺癌与25例乳腺纤维腺瘤组织中DNMT3a、3b蛋白的表达情况。甲基化特异性PCR检测108例散发性乳腺癌中BRCA1基因启动子甲基化状态。结果:与乳腺纤维腺瘤相比,乳腺癌组织中DNMT3a表达水平显著增高(P=0.037),DNMT3b表达水平增加不显著(P=0.478)。乳腺癌中BRCA1基因启动子存在甲基化。DNMT3a、3b过表达与乳腺癌病程晚期(TNMⅢ-Ⅳ期)显著相关(P=0.064,P=0.007);DNMT3a与BRCA1基因甲基化正相关(r=0.222,P=0.021),与BRCA1蛋白负相关(r=-0.172,P=0.022)。BRCA1蛋白表达与乳腺癌患者OS和DFS显著相关(P=0.025,P=0.027),DNMT3a表达与散发性乳腺癌OS和DFS存在一定的相关性(P=0.052,P=0.091)。结论:DNMT3a、3b高表达与乳腺癌侵袭转移、病程进展、预后相关。DNMT3a可能参与催化BRCA1基因启动子甲基化导致BRCA1蛋白表达下调的过程。抑制DNMT3a、3b蛋白可能有利于散发性乳腺癌的预防和治疗。
Objective: To investigate the association of DNMT3 a,3b expression with the state of promoter methylation of BRCA1 gene and BRCA1 expression in the development of sporadic breast cancer. Methods: We detected DNMT3 a,3b expression in 200 sporadic breast cancer tissues and 25 breast fibroadenoma tissues by immunohistochemistry. Using methylation specific PCR,we detected the state of promoter methylation of BRCA1 gene in 108 sporadic breast cancer patients. Results: Compared to breast fibroadenoma tissues,DNMT3 a expression was significantly increased( P = 0. 037) in breast tissues,but not DNMT3b( P = 0. 478). There was promoter methylation of BRCA1 gene in sporadic breast cancer patients. The overexpression of DNMT3 a and DNMT3 b was significantly correlated with the increased TNM stage of sporadic breast cancer patients( P = 0. 064,P = 0. 007). DNMT3 a was positively correlated with the state of BRCA1 gene promoter methylation( r = 0. 222,P = 0. 021) and negatively correlated with the state of BRCA1 expression( r =- 0. 172,P = 0. 022). BRCA1 expression was significantly correlated with OS or DFS of sporadic breast cancer patients( P = 0. 025,P = 0. 027),there was an correlation of DNMT3 a expression between OS or DFS of sporadic breast cancer patients in some degree( P = 0. 052,P = 0. 091). Conclusion: DNMT3 a and DNMT3 b overexpression is correlated with the invasion,metastasis and poor prognosis of sporadic breast cancer; DNMT3 a may participate in the progress of BRCA1 expression down- regulated regulated by BRCA1 gene promoter methylation. The inhibition of DNMT3 a and DNMT3 b have advantages in the progress of prevention and treatment of sporadic breast cancer.
Objective: To investigate the association of DNMT3 a,3b expression with the state of promoter methylation of BRCA1 gene and BRCA1 expression in the development of sporadic breast cancer. Methods: We detected DNMT3 a,3b expression in 200 sporadic breast cancer tissues and 25 breast fibroadenoma tissues by immunohistochemistry. Using methylation specific PCR,we detected the state of promoter methylation of BRCA1 gene in 108 sporadic breast cancer patients. Results: Compared to breast fibroadenoma tissues,DNMT3 a expression was significantly increased( P = 0. 037) in breast tissues,but not DNMT3b( P = 0. 478). There was promoter methylation of BRCA1 gene in sporadic breast cancer patients. The overexpression of DNMT3 a and DNMT3 b was significantly correlated with the increased TNM stage of sporadic breast cancer patients( P = 0. 064,P = 0. 007). DNMT3 a was positively correlated with the state of BRCA1 gene promoter methylation( r = 0. 222,P = 0. 021) and negatively correlated with the state of BRCA1 expression( r =- 0. 172,P = 0. 022). BRCA1 expression was significantly correlated with OS or DFS of sporadic breast cancer patients( P = 0. 025,P = 0. 027),there was an correlation of DNMT3 a expression between OS or DFS of sporadic breast cancer patients in some degree( P = 0. 052,P = 0. 091). Conclusion: DNMT3 a and DNMT3 b overexpression is correlated with the invasion,metastasis and poor prognosis of sporadic breast cancer; DNMT3 a may participate in the progress of BRCA1 expression down- regulated regulated by BRCA1 gene promoter methylation. The inhibition of DNMT3 a and DNMT3 b have advantages in the progress of prevention and treatment of sporadic breast cancer.
引文
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[13]Miki Y,Swensen J,Shattuck-Eidens D,et al.A strong candidate for the breast and ovarian cancer susceptibility gene BRCA1[J].Science,1994,266(5182):66-71.
[14]King MC,Marks JH,Mandell JB.Breast and ovarian cancer risks due to inherited mutations in BRCA1 and BRCA2[J].Science,2003,302(5645):643-646.
[15]Harvey JM,Clark GM,Osborne CK,et al.Estrogen receptor status by immunohistochemistry is superior to the ligand-binding assay for predicting response to adjuvant endocrine therapy in breast cancer[J].J Clin Oncol,1999,17(5):1474-1481.
[16]周建孟,袁建辉,姬娜娜,等.乳腺癌组织中DNA甲基化转移酶与多药耐药基因ABCG2表达的关系[J].癌变·畸变·突变,2009,21(03):194-200.
[17]Roll JD,Rivenbark AG,Jones WD,et al.DNMT3b overexpression contributes to a hypermethylator phenotype in human breast cancer cell lines[J].Mol Cancer,2008,7:15-18.
[2]Burgess M,Puhalla S.BRCA 1/2-mutation related and sporadic breast and ovarian cancers:More alike than different[J].Front Oncol,2014,4:19-22.
[3]Bestor TH,Ingram VM.Two DNA methyltransferases from murine erythroleukemia cells:Purification,sequence specificity,and mode of interaction with DNA[J].Proc Natl Acad Sci USA,1983,80(18):5559-5563.
[4]Okano M,Xie S,Li E.Cloning and characterization of a family of novel mammalian DNA(cytosine-5)methyltransferases[J].Nat Genet,1998,19(3):219-220.
[5]Pradhan S,Bacolla A,Wells RD,et al.Recombinant human DNA(cytosine-5)methyltransferase.Expression,purification,and comparison of de novo and maintenance methylation[J].J Biol Chem,1999,274(46):33002-33010.
[6]Okano M,Bell DW,Haber DA,et al.DNA methyltransferases DNMT3a and DNMT3b are essential for de novo methylation and mammalian development[J].Cell,1999,99(3):247-257.
[7]Saito Y,Kanai Y,Nakagawa T,et al.Increased protein expression of DNA methyltransferase(DNMT)1 is significantly correlated with the malignant potential and poor prognosis of human hepatocellular carcinomas[J].Int J Cancer,2003,105(4):527-532.
[8]Ding WJ,Fang JY,Chen XY,et al.The expression and clinical significance of DNA methyltransferase proteins in human gastric cancer[J].Dig Dis Sci,2008,53(8):2083-2089.
[9]Lin RK,Hsu HS,Chang JW,et al.Alteration of DNA methyltransferases contributes to 5'Cp G methylation and poor prognosis in lung cancer[J].Lung Cancer,2007,55(2):205-213.
[10]Daniel FI,Rivero ER,Modolo F,et al.Immunohistochemical expression of DNA methyltransferases 1,3a and 3b in oral leukoplakias and squamous cell carcinomas[J].Arch Oral Biol,2010,55(12):1024-1030.
[11]Amara K,Ziadi S,Hachana M,et al.DNA methyltransferase DNMT3b protein overexpression as a prognostic factor in patients with diffuse large B-cell lymphomas[J].Cancer Sci,2010,101(7):1722-1730.
[12]Rahman MM,Qian ZR,Wang EL,et al.DNA methyltransferases1,3a and 3b overexpression and clinical significance in gastroenteropancreatic neuroendocrine tumors[J].Hum Pathol,2010,41(8):1069-1078.
[13]Miki Y,Swensen J,Shattuck-Eidens D,et al.A strong candidate for the breast and ovarian cancer susceptibility gene BRCA1[J].Science,1994,266(5182):66-71.
[14]King MC,Marks JH,Mandell JB.Breast and ovarian cancer risks due to inherited mutations in BRCA1 and BRCA2[J].Science,2003,302(5645):643-646.
[15]Harvey JM,Clark GM,Osborne CK,et al.Estrogen receptor status by immunohistochemistry is superior to the ligand-binding assay for predicting response to adjuvant endocrine therapy in breast cancer[J].J Clin Oncol,1999,17(5):1474-1481.
[16]周建孟,袁建辉,姬娜娜,等.乳腺癌组织中DNA甲基化转移酶与多药耐药基因ABCG2表达的关系[J].癌变·畸变·突变,2009,21(03):194-200.
[17]Roll JD,Rivenbark AG,Jones WD,et al.DNMT3b overexpression contributes to a hypermethylator phenotype in human breast cancer cell lines[J].Mol Cancer,2008,7:15-18.