p53通路微小RNA在卵巢癌细胞和浆液性卵巢癌组织中的表达及意义  

Expression and significance of microRNAs in the p53 pathway in ovarian cancer cells and serous ovarian cancer tissues

作　　者：张旗[1] 何湘君[1] 马丽萍[1] 李娜[1] 杨静[1] 成夜霞[2] 崔恒[2] 

机构地区：[1]北京大学人民医院临床分子生物学研究所,100044 [2]北京大学人民医院临床分子生物学研究所妇科肿瘤中心,100044

出　　处：《中华肿瘤杂志》2011年第12期885-890,共6页Chinese Journal of Oncology

基　　金：国家自然科学基金（30740047）

摘　　要：目的探讨微小RNA（miRNA）miR-449a、miR-449b和miR-192家族是否与miR-34家族一样可作为p53通路成员，在卵巢癌中发挥作用。方法以DNA损伤药物阿霉素处理p53野生型卵巢癌A2780细胞，采用Westernblot法检测A2780细胞中p53蛋白的表达改变，采用实时定量PCR法检测激活p53后miR-449a／b、miR-34a、miR-34b、miR-34c、miR-192和miR-194的表达。在p53突变型卵巢癌SKOV3-ipl细胞中过表达miRNA，检测SKOV3．ipl细胞的细胞周期改变。采用实时定量PCR法检测miR-449a／b、miR-34a、miR-34b、miR-34c、miR-192和miR-194在正常输卵管以及不同分级分期的浆液性卵巢癌组织中的表达。结果在阿霉素处理后24h，卵巢癌A2780细胞中p53蛋白的表达明显升高，miR-34b、miR-34c、miR-449a／b的表达约升高19～21倍，但miR-192和miR-194的表达未见明显变化。转染miR449b和miR-34c后，SKOV3．ipl细胞出现G，期阻滞。miR-449a／b在浆液性卵巢癌组织中的表达改变与miR-34b和miR-34c基本平行，它们在高级别和晚期浆液性卵巢癌组织中的表达水平，显著低于正常输卵管组织以及低级别和早期浆液性卵巢癌组织；miR-192、miR-194和miR-34a在卵巢癌组织中的表达未发现明显的特征，在正常输卵管组织中的表达也明显低于miR-449a／b、miR-34b和miR-34e。结论miR-449a／b与miR-34b、miR-34e作为抑癌基因，在p53通路中发挥协同作用，它们的功能缺失与浆液性卵巢癌的发生、发展密切相关。Objective The aim of this study was to investigate whether miR-449a, miR-449b and miR-192 family microRNAs play the same roles in p53 pathway as miR-34 family in ovarian cancer. Methods Wild-type p53 ovarian carcinoma cell line A2780 cells were treated with genotoxic agent adriamycin. The reactivation of p53 was detected by Western blot. The expression of miR-449a/b, miR- 34a, miR-34b,miR-34c, miR-192 and miR-194 were detected by real-time quantitative PCR. Mutant p53 ovarian cancer cell line SKOV3. ipl cells were transfected with pre-microRNAs and the cell-cycle changes were detected. The expression level of miR-449a/b, miR-34a, miR-34b, miR-34c, miR-192 and miR-194 in serous ovarian carcinomas of varying grade and stage were compared with real-time PCR. Results The expressions of miR-449a/b, miR-34b and miR-34c were 19-fold to 21-fold elevated after p53 activation by genotoxic agent. Ectopic expression of miR-449b, as well as miR-34c, resulted in cell-cycle arrest in SKOV3. ipl cells： The expression of miR-449a/b was parallel with that of miR-34b, miR-34C, and Were significantly lower in late stage and high-grade serous carcinomas than in the normal fallopian tube, early stage and low-grade serous carcinomas. The expression of miR-192, miR-194 and miR-34a did not show evident features in serous ovarian carcinomas and were much lower than miR-449a/b, miR-34b and miR-34c in normal fallopian tube. Conclusions As tumor-suppressor microRNAs, miR-449a/b, miR-34b and miR- 34c cooperate and play important roles in p53 pathway. Their inactivation may contribute to the carcinogenesis and progression of serous ovarian carcinomas.

关 键 词：微小RNA P53通路 卵巢肿瘤 

分 类 号：R737.31[医药卫生—肿瘤]