miR-410 regulates apoptosis by targeting Bak1 in human colorectal cancer cells
miR-410 regulates apoptosis by targeting Bak1 in human colorectal cancer cells
复制标题
DOI：
10.3892/mmr.2016.5271
复制
发表时间：
2016-07-01
影响因子：
3.4
通讯作者：
Gao, Yang
中科院分区：
文献类型：
--
作者：
Liu, Chunyuan;Zhang, Aihong;Gao, Yang
MicroRNAs(MiRs)在结直肠癌(CRC)的发病机制中起重要作用。已有研究表明miR-410在肿瘤中具有多方面的作用，但其是否影响结直肠癌细胞的凋亡尚不清楚。为了证明miR-410在结直肠癌中的作用，本研究检测了miR-410在结直肠癌组织和细胞系中的表达，并通过在结直肠癌细胞中导入miR-410或miR-410抑制剂来调控miR-410的水平。用四甲基偶氮唑盐比色法、Western印迹法和细胞色素C法检测细胞的生长和凋亡。通过荧光素酶活性测定进行靶向验证。研究发现miR-410在结直肠癌组织和细胞系中表达上调。MiR-410过表达导致细胞生长活性增强，细胞凋亡程度降低。相反，内源性miR-410的抑制激活了细胞的凋亡机制。Western印迹分析和荧光素酶活性分析表明，Bak1直接被miR-410靶向，并且Bak1基因的敲除减弱了miR-410抑制的促凋亡作用。此外，研究还发现Bak1在结直肠癌组织中的表达下调，并且与miR-410的表达呈负相关，这进一步支持了Bak1受miR-410的调控。本研究结果提示miR-410可能通过抑制细胞凋亡的基础水平而发挥致癌miR的作用。这些发现可能有助于理解癌症发生的分子机制。
MicroRNAs (miRs) are essential in the pathogenesis of colorectal cancer (CRC). Previous studies have demonstrated that miR-410 exerts multiple effects on tumors, however, whether it affects the apoptosis of CRC cells remains to be elucidated. In the present study, to demonstrate the role of miR-410 in CRC, miR-410 expression was detected in CRC tissues and cell lines, and the miR-410 level was manipulated by transfection with an miR-410 or miR-410 inhibitor in CRC cells. Cell growth and apoptosis was tested using an MTT assay, western blot and cytochrome C assay. Target validation was conducted by luciferase assay. It was found that miR-410 was upregulated in CRC tissues and cell lines. The overexpression of miR-410 resulted in an increase in growth activity and decrease in the extent of apoptosis. By contrast, the inhibition of endogenous miR-410 activated the apoptotic machinery. Western blot analysis and a luciferase activity assay showed that Bak1 was directly targeted by miR-410, and that knockdown of Bak1 attenuated the pro-apoptotic effect of miR-410 inhibition. In addition, it was shown that the expression of Bak1 was downregulated in CRC tumor tissues and was reversely correlated with the expression of miR-410, which provided further support that Bak1 was regulated by miR-410. The results of the present study suggested that miR-410 may function as an oncogenic miR by suppressing the basal level of apoptosis. These findings may assist in understanding the molecular mechanisms of cancer development.