目的 旨在探讨miR-449b-5p在肝细胞癌发病机制中的作用.方法 用miR-449b-5p模拟物或miR-449b-5p抑制剂转染肝细胞癌细胞系.通过CCK-8和Transwell实验鉴定mi R-449b-5p对肝细胞癌细胞增殖和迁移能力的影响.Westernblot和qRT-PCR检测miR-449b-5p和KLF4m RNA的表达水平.采用双荧光素酶法验证miR-449b-5p与KLF4的关系.结果 研究结果显示miR-449b-5p在人肝癌组织和细胞系中的表达上调.此外, 降低miR-449b-5p的表达抑制了肝癌细胞的增殖和迁移能力.机制方面, 实验数据证明KLF4是miR-449b-5p在肝癌细胞中的直接作用靶点.同时, 拯救实验证实过表达KLF4可以抵消miR-449b-5p促进肝癌细胞增殖和转移的能力.与此同时, 研究结果表明细胞周期抑制剂p21在HCC细胞中异常下调, 而降低mi R-449b-5p表达后这种抑制作用被逆转.结论 研究结果表明miR-449b-5p可通过靶向KLF4促进肝癌细胞的增殖和转移.%Objective This study was designed to explore the role of miR-449b-5p in the pathogenesis of hepatocellular carcinoma. Methods HCC cell lines were transfected with miR-449b-5p mimics or miR-449b-5p inhibitor. Proliferation and migration were identified by MTT assay and transwell assay, respectively. Protein and mRNA expression levels of associated genes were measured by Western blot and quantitative real-time PCR (qRT-PCR) , respectively. Dual luciferase assay was used to determine the relation of miR-449b-5p and KLF4. Results We showed that miR-449b-5p expression was upregulated in human HCC tissues and cell lines. Decreased expression of miR-449b-5p inhibited the HCC cell proliferation and migration. KLF4 was identified as a direct target of miR-449b-5p in HCC cells. Furthermore, overexpression of KLF4 attenuated the effects of miR-449b-5p on the regulation of HCC cell motility. We also showed that the cell cycle inhibitor p21 was aberrantly downregulated in HCC cells, whereas this inhibition was reversed by miR-449b-5p inhibitor. Conclusion Our findings illuminated miR-449b-5p targeting KLF4 to promote the proliferation and metastasis in HCC cells.
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