Zhang Qi, He Xiang-jun, Ma Li-ping, Li Na, Yang Jing, Cheng Ye-xia, Cui Heng
Institute of Clinical Molecular Biology, Peking University People's Hospital, Beijing 100044, China.
Zhonghua Zhong Liu Za Zhi. 2011 Dec;33(12):885-90.
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.
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.
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.
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.
本研究旨在探究miR - 449a、miR - 449b和miR - 192家族微小RNA在卵巢癌中是否与miR - 34家族在p53通路中发挥相同作用。
用基因毒性药物阿霉素处理野生型p53卵巢癌细胞系A2780细胞。通过蛋白质印迹法检测p53的重新激活情况。通过实时定量PCR检测miR - 449a/b、miR - 34a、miR - 34b、miR - 34c、miR - 192和miR - 194的表达。用前体微小RNA转染突变型p53卵巢癌细胞系SKOV3.ipl细胞并检测细胞周期变化。通过实时PCR比较不同分级和分期的浆液性卵巢癌中miR - 449a/b、miR - 34a、miR - 34b、miR - 34c、miR - 192和miR - 194的表达水平。
经基因毒性药物激活p53后,miR - 449a/b、miR - 34b和miR - 34c的表达升高了19倍至21倍。miR - 449b以及miR - 34c的异位表达导致SKOV3.ipl细胞出现细胞周期阻滞。miR - 449a/b的表达与miR - 34b、miR - 34c的表达平行,在晚期和高级别浆液性癌中的表达明显低于正常输卵管、早期和低级别浆液性癌。miR - 192、miR - 194和miR - 34a在浆液性卵巢癌中未表现出明显特征,且在正常输卵管中的表达远低于miR - 449a/b、miR - 34b和miR - 34c。
作为肿瘤抑制性微小RNA,miR - 449a/b、miR - 34b和miR - 34c协同作用,在p53通路中发挥重要作用。它们的失活可能有助于浆液性卵巢癌的发生和发展。