miR-144靶向PIK3CD基因影响结直肠癌细胞增殖和细胞因子表达的作用机制

目的:探究miR-144与磷脂酰肌醇-3肌酶催化亚基δ(PIK3CD)的靶向关系及其影响结直肠癌(CRC)细胞增殖和免疫功能的作用机制。方法:RT-qPCR和Westernblot检测CRC肿瘤组织以及HCT116细胞中miR-144和PIK3CD表达水平的变化。miRcode在线预测miR-144和PIK3CD的靶向关系,使用双荧光素酶实验进行验证。将miR-144模拟物、抑制剂和PIK3CDsiRNA分别转染至HCT116细胞后,RT-qPCR和Westernblot检测miR-144以及PIK3CDmRNA和蛋白表达水平的变化,MTT法检测细胞增殖能力的变化,ELISA法检测TNF-α和sIL-2R表达水平的变化。结果:在CRC肿瘤组织和HCT116细胞中,miR-144表达下调,PIK3CD表达上调。PIK3CDsiRNA和miR-144模拟物能够促使PIK3CDmRNA和蛋白表达下调,抑制HCT116细胞增殖,同时促进TNF-α和sIL-2R表达下调;miR-144抑制剂作用与之相反。结论:miR-144通过靶向PIK3CD基因抑制HCT116细胞增殖及其免疫功能。

Effect mechanism of miR-144 on proliferation and immune function of colorectal cancer cells by targeting PIK3CD gene

Objective: To explore the target relationship between miR-144 and PIK3CD and their mechanism of proliferation and immune function in colorectal cancer cells. Methods: RT-qPCR and Western blot were used to detect the expression of miR-144 and PIK3CD in colorectal tumor tissue and HCT116 cells.The adual luciferase reporter assay was used to confirm the target relationship which predicted by miRcode software between miR-144 and PIK3CD.Next,miR-144 mimics,inhibitors and PIK3CD siRNA were transfected into HCT116 cells respectively;RT-qPCR and Western blot were used to detect the mRNA and protein expression level of miR-144 and PIK3CD;MTT was performed to detect the of cell proliferation;ELISA was used to detect the expression of TNF-α and sIL-2R. Results: In CRC tumor tissues and HCT116 cells,miR-144 was down-regulated and PIK3CD was up-regulated.PIK3CD siRNA and miR-144 mimics could reduce the mRNA and protein expression of PIK3CD,inhibit cell proliferation and promote the expression of TNF-α and sIL-2R.While the effect of miR-144 inhibitor was opposite. Conclusion: miR-144 could inhibit proliferation and immune function of HCT116 cells by targeting PIK3CD gene.