Overexpression of microRNA-30a-5p inhibits liver cancer cell proliferation and induces apoptosis by targeting MTDH/PTEN/AKT pathway
Overexpression of microRNA-30a-5p inhibits liver cancer cell proliferation and induces apoptosis by targeting MTDH/PTEN/AKT pathway
复制标题
microRNA-30a-5p过表达通过靶向MTDH/PTEN/AKT通路抑制肝癌细胞增殖并诱导细胞凋亡
DOI：
10.1007/s13277-015-4456-1
复制
发表时间：
2016-05-01
期刊：
影响因子：
--
通讯作者：
Liu, Chang-an
中科院分区：
文献类型：
--
作者：
Li, Wen-fang;Dai, Hang;Liu, Chang-an
越来越多的结果表明，microRNAs(MiRNAs)在恶性肿瘤的发展过程中可能发挥重要作用。MiR-30a-5p在多种癌症中表达下调，是一种抑癌基因。MiRNA-30a-5p在肝细胞癌中的作用和分子机制尚不清楚。本研究采用实时定量聚合酶链式反应(qRT-PCR)检测了16对肝癌及其癌旁组织和肝癌细胞系中miR-30a-5p的表达。通过过表达miRNA-30a-5p、CCK-8和结肠形成实验检测细胞生长，用流式细胞仪检测细胞凋亡。Western印迹法检测蛋白表达。并通过荧光素酶活性测定分析其可能的作用机制。裸鼠体内观察肝癌细胞株的生长情况。结果表明，miR-30a-5p在肝癌组织中的表达明显低于癌旁肝组织，在肝癌细胞系中的表达也低于正常肝细胞。荧光素酶分析表明，胃黏附素(Mtdh)是miR-30a-5p的直接靶标。肝癌组织中miR-30a-5p与MTDH值呈显著负相关。在肝癌细胞中过表达miR-30a-5p显著抑制细胞增殖、结肠癌形成和诱导细胞凋亡，而MTDH过表达则逆转了miRNA-30a-5p在肝癌细胞中的生长抑制和诱导凋亡。MiRNA-30a-5p上调磷酸酶和张力蛋白同源蛋白(PTEN)的表达，从而通过靶向MTDH抑制AKT的激活。MiRNA-30a-5p在体内也显著抑制了HepG2肿瘤的生长。我们的结果表明，miR-30a-5p的低表达可能通过直接靶向mTdH来抑制肝癌中的miRNA，因此是一种潜在的肝癌靶向治疗的候选生物标记物。
Increasing results suggest microRNAs (miRNAs) could function important roles in malignant tumor progression. miR-30a-5p is downregulated in variety of cancers and acts as a cancer suppressing gene. The functions and molecular mechanisms of miRNA-30a-5p in hepatocellular carcinoma (HCC) remain unclear. In the present study, quantitative real-time PCR (qRT–PCR) was used to detect miR-30a-5p expression in 16 pairs of HCC and their adjacent non-cancerous tissues and HCC cell lines. By overexpression of miRNA-30a-5p, CCK-8 and colon formation assay were used to evaluate cell growth and flow cytometry to evaluate cell apoptosis. Western blot was used to test protein expression. And potential mechanisms were analyzed with luciferase activity assay. In vivo HepG2 tumor growth was observed with nude mice. Our results showed that miR-30a-5p expression in HCC tissues was significantly lower compared to adjacent non-cancerous liver tissues, and lower miR-30a-5p expression was also observed in HCC cell lines compared to normal liver cell. Luciferase assay showed that metadherin (MTDH) mRNA was a direct target of miR-30a-5p. A significant reverse correlation between miR-30a-5p and MTDH in liver cancer tissues was observed. miR-30a-5p overexpression in HCC cells significantly inhibited cell proliferation, colon formation, and induced apoptosis while MTDH overexpression reversed growth inhibition and apoptosis induction of miRNA-30a-5p in HCC cells. miRNA-30a-5p upregulated phosphatase and tensin homolog (PTEN) protein expression and thus inhibited AKT activating by targeting MTDH. miRNA-30a-5p also significantly inhibited HepG2 tumor growth in vivo. Our results suggest that underexpression of miR-30a-5p might function as a tumor suppressing miRNA by directly targeting MTDH in HCC and is therefore a potential candidate biomarker for HCC targeting therapy.