HPV16 E7蛋白阳性和阴性乳癌组织中P16蛋白与p16基因启动子区甲基化检测  

Detection of P16 protein and methylation status of promoter region of p16 gene in HPV16 E7 protein-positive and-negative breast carcinoma tissue

作　　者：李新娟[1] 崔静[2] 朱慧芳[2] 韩芳毅[2] 杨慧林[2] 王霞[2] 冶亚平[2] 李新强[2] 马帅[2] 张洁[2] 贺国洋[2] 赵杰[2] 常海敏[3] 千新来[2] 

机构地区：[1]新乡医学院生理学教研室,新乡453003 [2]新乡医学院病理学教研室,新乡453003 [3]新乡医学院法医临床学教研室,新乡453003

出　　处：《郑州大学学报（医学版）》2009年第4期729-731,共3页Journal of Zhengzhou University(Medical Sciences)

基　　金：卫生部科研基金资助项目WKJ2007-2-022;河南省高校杰出科研人才创新工程资助基金项目2007KYCX013

摘　　要：目的:研究人乳头瘤病毒16型(HPV16)E7蛋白阳性(E7+)和阴性(E7-)乳癌组织中P16蛋白表达与p16基因启动子区甲基化状态的关系,以揭示HPV16E7+乳癌组织中P16蛋白高表达的可能机制。方法:以经PCR筛选证实的HPV16DNA阳性的56例乳癌组织(经Westernblotting证实其中42例HPV16E7+,14例E7-)为研究对象。采用甲基化特异性聚合酶链反应(MSP)方法检测癌组织中p16基因启动子区甲基化状态;采用Westernblotting方法测定P16蛋白。结果:42例HPV16E7+乳癌组织中,p16基因启动子区甲基化率为11.9%(5/42),P16蛋白阳性率85.7%(36/42),2者表达密切相关(rp=0.912,P<0.001)。14例HPV16E7-乳癌组织中,p16基因启动子区甲基化率为78.0%(11/14),P16蛋白阳性率21.4%(3/14),2者表达有较密切的关系(rp=0.779,P=0.043)。HPV16E7+乳癌组织中p16基因启动子区甲基化率低于HPV16E7-乳癌组织(χ2=17.405,P<0.01),P16蛋白阳性率高于HPV16E7-乳癌组织(χ2=20.525,P<0.01)。结论:HPV16E7蛋白可能通过降低p16基因的甲基化使P16蛋白表达水平升高,从而在乳癌的发生、发展过程中发挥重要作用。Aim： To investigate the relationship between the expression of P16 protein and methylation status of promotet region of p16 gene in human papillomavirus 16 type （HPV16） E7 protein-positive （E7＋） and -negative （E7-） breast carcinoma tissue. Methods：A total of 56 HPV16 DNA-positive breast carcinoma specimens, including 42 HPV16 E7 ＋ and 14 HPV16 E7 - breast carcinoma specimens certified by Western blotting assay,were subjected to detect the methylation status of p16 promoter region by methylation-specific polymerase chain reaction （MSP） and P16 protein by Western blotting assay. Results： The methylation rate of p16 promoter region in HPV16 E7＋ group was 11.9% （5/42） , the positive rate of P16 was 85.7% （36/42） , and the expression of P16 was closely associated with the methylation status of p16 promoter region （rp = 0.912,P = 0. 000）. The methylation rate of p16 promoter region in HPV16 E7- group was 78.6% （11/ 14） , the positive rate of P16 was 21.4% （3/14） , and the expression of P16 was associated with the methylation status of p16 promoter region （ rp = 0. 779,P = 0. 043 ）. Compared with those （χ^2 = 20. 525, P 〈 0.01 ） in HPV16 E7 - group, the methylaiion rate of p16 promoter region was lower （χ^2 = 17. 405, P 〈 0.01 ） in HPV16 E7 ＋ group （ P 〈 0.01 ）. Conclusion ： Hypomethylation status may be the foundation of p16 transcription activated by the regulatory feedback in pRb/p16 pathway due to the removal of the Rb repression by E7.

关 键 词：人乳头瘤病毒16型 P16基因 启动子 甲基化 乳癌 

分 类 号：R737.9[医药卫生—肿瘤]