miR-135b对结肠癌细胞凋亡诱导作用及机制  

Effect and mechanism of miR-135b on apoptosis of HCT116 colon cancer cells

作　　者：张小鸿 黄枝妙 朱颖[1] 袁平[1] ZHANG Xiao-hong;HUANG Zhi-miao;ZHU Ying(Fujian Provincial Center for Disease Control and Prevention,Fuzhou,Fujian Province 350001,China)

机构地区：[1]福建省疾病预防控制中心,福州350001

出　　处：《中国公共卫生》2022年第6期783-786,共4页Chinese Journal of Public Health

基　　金：福建省卫生健康科技计划项目资助(2018-1-23,2015-ZQN-JC-9)。

摘　　要：目的探讨miR-135b对HCT116结肠癌细胞凋亡的诱导作用及机制。方法HCT116细胞分为对照组、miR-135b NC组及miR-135b inhibitor组,噻唑蓝(MTT)法检测细胞活力,Hoechst检测细胞凋亡,Western blot法检测B淋巴细胞瘤-2基因(Bcl-2)、Bcl-2相关蛋白X(Bax)、多聚ADP核糖多聚酶(cleaved-PARP)、含半胱氨酸的天冬氨酸蛋白水解酶(cleaved-caspase 3)、人第10号染色体缺失的磷酸酶及张力蛋白同源基因(PTEN)、磷脂酰肌醇3-激酶(PI3K)、蛋白激酶A(AKT)表达。结果与对照组(1.00±0.03)及miR-135b NC组(1.02±0.04)比较,miR-135b inhibitor组细胞中miR-135b表达(0.53±0.05)显著下调(F=259.19,P<0.05)。对照组、miR-135b NC组、miR-135b inhibitor组HCT116细胞活力和细胞凋亡率分别为(0.58±0.05)、(0.59±0.06)、(0.39±0.04)和(3.24±0.13)%、(3.27±0.08)%、(36.48±0.52)%,与对照组、miR-135b NC组比较,miR-135b inhibitor组HCT116细胞活力明显下降(P<0.05),凋亡率显著升高(P<0.05)。与对照组、miR-135b NC组比较,miR-135b inhibitor组HCT116细胞Bcl-2、cleaved-caspsase 3、PI3K及p-AKT蛋白表达下调(P<0.05),Bax、cleaved-PARP及PTEN蛋白表达明显上调(P<0.05)。结论下调miR-135b可诱导HCT116细胞凋亡,其机制可能与阻断PTEN/PI3K/AKT信号通路有关。Objective To explore the effect and mechanism of microRNA-135b(miR-135b)on apoptosis of HCT116colon cancer cells.Methods HCT116 cells were divided into control group,miR-135b NC group and miR-135b inhibitor group.Cell viability was detected with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assay.Cell apoptosis was detected with Hoechst staining.The expression of B-cell lymphoma-2(Bcl-2),Bcl-2-associated X(Bax),poly ADP-ribose polymerase(cleaved-PARP),cysteinyl aspartate specific proteinase(cleaved-caspase 3),phosphatase and tensin homolog deleted on chromosome ten(PTEN),phosphatidylinositol 3-kinase(PI3K),and protein kinase A(AKT)were detected with Western blot.Results The expression of miR-135b was down-regulated in miR-135b inhibitor group compared with that in control group and miR-135b NC group(0.53±0.05 vs.1.00±0.03 and 1.02±0.04)(F=259.19,P<0.05).The cell viability and apoptotic rate of HCT116 cells were 0.58±0.05 and 3.24±0.13% for control group,0.59±0.06 and 3.27±0.08% for miR-135b NC group,and 0.39±0.04 and 36.48±0.52% for miR-135b inhibitor group,respectively,with significantly decreased viability but increased apoptotic rate of HCT116 cells for miR-135b inhibitor group compared with that for control group and miR-135b NC group(both P<0.05).Significantly down-regulated expressions of Bcl-2,cleaved-caspase 3,PI3K,and p-Akt and up-regulated expressions of Bax,cleaved-PARP,and PTEN were detected in miR-135b inhibitor group in comparison with those in control group and miR-135b NC group was(P<0.05 for all).Conclusion Dow-regulation of miR-135b could induce apoptosis of HCT116 cells,which might be related to the blocking of PTEN/PI3K/Akt signaling pathway.

关 键 词：miR-135b 结肠癌 HCT116细胞 凋亡 

分 类 号：R735.35[医药卫生—肿瘤]