卵巢上皮恶性肿瘤侵袭转移相关miRNA的筛选与鉴定

目的 探索与卵巢上皮恶性肿瘤侵袭转移可能相关的miRNA.方法 采用miRNA芯片,筛选SKOV-3ip和SKOV-3细胞差异表达miRNA.利用生物信息学软件TargetScan、MicroCosm、PicTar和GO,预测差异表达miRNA的靶基因及其功能.采用实时逆转录聚合酶链反应(real-time RT-PCR)技术,验证与卵巢癌侵袭转移可能相关的5种miRNA(let-7a、let-7e、let-7f、miR-22和miR-886-5p)在SKOV-3ip和SKOV-3细胞的表达.同时,检测这5种miRNA在另外一组侵袭转移能力不同的卵巢癌细胞株HO8910和HO-8910PM中的表达,并进行统计学分析.结果 基因芯片筛选显示,42种miRNA在SKOV-3ip和SKOV-3细胞株表达差异明显.进一步分析显示,let-7a、let-7e、let-7f、miR-22和miR-886-5p等5种miRNA可能与卵巢癌的侵袭转移密切相关.real-time RT-PCR结果证实,let-7f和miR-22在两组侵袭转移能力不同的卵巢癌细胞(SKOV-3和SKOV-3ip细胞、HO-8910和HO-8910PM细胞)表达差异有统计学意义(均P＜0.05).结论 let-7f和miR-22在侵袭转移能力强的卵巢癌细胞中低表达,可能具有抑癌基因的作用.

Screening and identification of potential miRNA involved in ovarian cancer invasion and metastasis

Objective To identify potential miRNA involved in epithelial ovarian cancer (EOC)invasion and metastasis. Methods miRNA microarray was applied to compare the miRNA expression profile between SKOV-3ip and SKOV-3 cells. Bioinformatics programs (TargetScan, MicroCosm, PicTar and GO) were used to analyze the miRNA and their potential target genes. Real-time RT-PCR was used to confirm the results of microarray and for expanding detection in another paired EOC cell lines (HO-8910 and HO-8910PM). Results Totally, expressions of 42 miRNA were found significantly different between SKOV-3ip and SKOV-3 cells. Among them, 10 miRNA were down-regulated, including let-7a, let-7f, miR22 and miR-886-5p; while 32 were up-regulated, for example, let-7e and miR-519e. Bioinformatic analysis indicated that let-7a, let-7e, let-7f, miR-22 and miR-886-5p may be involved in cancer invasion and metastasis. Meanwhile, real-time RT-PCR confirmation and statistic analysis showed that let-7f and miR-22expressions were significantly different between ovarian cancer cell lines with various invasive and metastatic capacity (P ＜ 0.05). Conclusion The expression of let-7f and miR-22 is low in ovarian cancer cells with high invasive and metastatic capacity. It suggests that they are potential tumor suppressor genes. Further research on their role and mechanism is needed.