Objective: To investigate the clinical implications of abnormal expression of miR-607 in hepatocellular carcinoma (HCC) and its influence on HCC cell proliferation and migration.
Methods: The expression of miR-607 in 45 pairs of HCC and adjacent tissues were detected with real-time PCR, and the correlation between miR-607 expression and clinicopathological features of the patients was analyzed. The effects of transfection with miR-607 mimics on proliferation, apoptosis, migration and invasion of two HCC cell lines (Huh-7 and HCCLM3) were evaluated using CCK-8 assay, flow cytometry, wound-healing assay and Transwell assay. A dual-luciferase reporter system was used to detect the direct binding between miR-607 and 3'-UTR of TRPC5 mRNA. Western blotting was used to measure the expressions of TRPC5, CCND1, MMP2 and phosphorylated Akt in the HCC cells.
Results: The expression of miR-607 was significantly decreased in HCC tissues (P=0.029) and HCC cell lines (P < 0.05). In HCC patients, a low expression of miR-607 was associated with a larger tumor size (>5 cm, P=0.031), vascular invasion (P=0.027) and advanced TNM stages (Ⅲ + Ⅳ, P=0.015). In the two HCC cell line, overexpression of miR-607 significantly inhibited cell proliferation, migration, and invasion and enhanced cell apoptosis (P < 0.05). The results of dualluciferase reporter assay confirmed that TRPC5 was a direct target of miR- 607 in HCC cells. Overexpression of miR-607 significantly inhibited the expressions of TRPC5, CCND1, and MMP2 and suppressed Akt phosphorylation in HCC cells (P < 0.05).
Conclusion: A low expression of miR-607 in HCC is associated with poor clinicopathological phenotypes of HCC. Overexpression of miR-607 inhibits HCC growth and metastasis possibly by down- regulating TRPC5, which causes Akt signaling inactivation.
目的: 研究miR-607异常表达在肝细胞癌（HCC）中的临床意义及对肝癌细胞生长转移的影响。
方法: 荧光定量PCR检测45对HCC及癌旁组织中miR-607表达，分析miR-607表达与患者临床病理特征间的相关性。通过转染miR-607 mimics提高HCC细胞（Huh-7和HCCLM3）内miR-607的表达水平。采用CCK-8、流式细胞仪、划痕愈合实验及transwell小室模型分别检测过表达miR-607对HCC细胞增殖、凋亡、迁移和侵袭的影响。双荧光素酶报告基因系统检测miR-607是否与潜在靶点TRPC5的mRNA 3'-非翻译区直接结合。Western blot检测过表达miR-607对HCC细胞内TRPC5、CCND1、MMP2表达及Akt磷酸化的影响。
结果: MiR-607在HCC组织及HCC细胞中的表达水平均降低（P < 0.05）；miR-607低表达与肿瘤直径>5 cm、血管侵犯及较晚TNM分期（Ⅲ+Ⅳ）密切相关（P < 0.05）。过表达miR-607抑制HCC细胞增殖、迁移、侵袭并诱导细胞凋亡（P < 0.05）。双荧光素酶报告基因系统证实miR-607与TRPC5 mRNA 3'-非翻译区直接结合（P < 0.05）。过表达miR-607抑制HCC细胞内TRPC5、CCND1及MMP2表达并下调Akt的磷酸化水平（P < 0.05）。
结论: miR-607低表达与肝细胞癌恶性临床特征密切相关，miR-607可能通过下调TRPC5表达进而抑制Akt通路激活来发挥抗HCC生长转移作用。
Keywords: Akt signal pathway; TRPC5; hepatocellular carcinoma; metastasis; miR-607.