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  

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

作　　者：陈红跃[1] 杨萌萌[2] 张小路[3] 段铮[3] CHEN Hong-Yue;YANG Meng-Meng;ZHANG Xiao-Lu;DUAN Zheng(Department of General Surgery,Henan Province Hospital of TCM,Zhengzhou 450000,China)

机构地区：[1]河南省中医院普外科,郑州450000 [2]河南中医药大学第二附属医院,郑州450000 [3]河南省中医院肿瘤科,郑州450000

出　　处：《中国免疫学杂志》2019年第10期1179-1183,1187,共6页Chinese Journal of Immunology

基　　金：河南省中医药科学研究专项项目(2016ZY2063)资助

摘　　要：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。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.

关 键 词：miR-144 PIK3CD 结肠直肠癌 增殖 免疫 

分 类 号：R392.1[医药卫生—免疫学]