miR-486 regulates metastasis and chemosensitivity in hepatocellular carcinoma by targeting CLDN10 and CITRON
miR-486 regulates metastasis and chemosensitivity in hepatocellular carcinoma by targeting CLDN10 and CITRON
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miR-486通过靶向CLDN10和CITRON调节肝细胞癌的转移和化疗敏感性
DOI：
10.1111/hepr.12500
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发表时间：
2015-12-01
影响因子：
4.2
通讯作者：
Wang, Lisheng
中科院分区：
文献类型：
--
作者：
Sun, Huiyan;Cui, Chunping;Wang, Lisheng
目的：miRNA-486（miR-486）是从人胎肝cDNA文库中首次发现的，被认为与肝细胞癌（HCC）的发生发展有关。方法：采用实时荧光定量PCR方法检测miR-486在肝癌组织、细胞系和血清中的表达，并与肿瘤转移和化疗敏感性的关系。通过慢病毒转染在细胞系SMMC-7721/LM 3中进行miR-486过表达或下调。使用商业测定定量细胞增殖、迁移和凋亡。明胶酶谱法测定基质金属蛋白酶活性。结果：miR-486在肝癌组织和细胞系中表达下调，miR-486在肝癌组织和细胞系中表达下调的频率较高，miR-486在肝癌组织和细胞系中表达下调的频率较高。慢病毒介导的miR-486在HCC细胞系中的恢复导致细胞生长、集落形成和迁移能力的显著降低。然而，miR-486抑制增强HCC细胞的增殖和侵袭。分别调节细胞增殖和侵袭的两个基因CITRON和CLDN 10被鉴定为HCC细胞中miR-486的直接靶点。通过siRNA敲低CITRON和CLDN 10导致HCC细胞系中miR-486恢复的相似表型。结论：miR-486可能是一种新的肝癌抑制基因。
Aim: miRNA-486 (miR-486) was first identified from the human fetal liver cDNA library and considered to be associated with hepatocellular carcinoma (HCC) development. Its roles in regulation of HCC metastasis and chemosensitivity have not been explored yet.Methods: miR-486 expression in HCC tissues, cell lines and serum was evaluated by real-time polymerase chain reaction. miR-486 overexpression or downregulation in the cell lines SMMC-7721/LM3 was conducted by lentivirus transfection. Cell proliferation, migration and apoptosis were quantitated using commercial assays. Matrix metalloproteinase activity was quantitated by gelatin zymography. The target genes of miR-486 were screened by 3'-untranslated region luciferase report assays and their function was validated by small RNA interference.Results: We show here that miR-486 is frequently down-expressed in HCC tissues and cell lines. Lentivirus-mediated restoration of miR-486 in HCC cell lines resulted in significant reduction in the ability of cell growth, colony formation and migration. However, miR-486 inhibition enhances proliferation and invasion of HCC cells. Two genes, CITRON and CLDN10 which regulate cell proliferation and invasion, respectively, were identified as the direct targets of miR-486 in HCC cells. CITRON and CLDN10 knockdown by siRNA results in similar phenotypes of miR-486 restoration in HCC cell lines. In addition, miR-486 enhances the chemosensitivity of HCC cells to sorafenib.Conclusion: Our data indicated that miR-486 may function as a novel tumor suppressor in HCC.