缺氧条件下miR-23c靶向ZNF420调控非小细胞肺癌细胞A549的凋亡

MiR-23c targets ZNF420 to regulate apoptosis of non-small cell lung cancer cell line A549 under hypoxia

目的探讨缺氧条件下miR-23c靶向调控ZNF420对非小细胞肺癌细胞A549凋亡的影响。方法过表达miR-23c后,用Annexin-V染色观察在缺氧或不缺氧的条件下非小细胞肺癌细胞A549的凋亡情况;TargetScan在线分析miR-23c与ZNF420的相关性:其次将ZNF420的3端非编码区克隆进pmiR-RB-Repor Luciferase载体,利用双荧光素酶报告系统检测miR-23c是否靶向调控ZNF420;在过表达或敲低miR-23c的情况下,通过Western Blot检测ZNF420蛋白表达量和细胞凋亡标志蛋白的表达量。结果过表达miR-23c时,缺氧环境下,非小细胞肺癌细胞A549凋亡水平显著上升(P<0.05)。miR-23c靶向ZNF420的3′UTR的67-74的位置。过表达miR-23c时,ZNF420的表达量显著下降(P<0.05),细胞凋亡标志蛋白cleaved-caspase9、BAX表达量上升(P<0.05),Bcl-2表达量下降(P <0.05)。敲低miR-23c时,ZNF420的表达量显著上升(P<0.05),细胞凋亡标志蛋白cleaved-caspase9、BAX表达量下降(P<0.05),Bcl-2表达量上升(P<0.05)。结论缺氧条件下miR-23c通过靶向ZNF420的mRNA调控ZNF420的表达量来促进非小细胞肺癌细胞A549的凋亡。

Objective To investigate the effect of miR-23 c targeting ZNF420 on apoptosis of non-small cell lung cancer cell line A549 under hypoxia.Methods After overexpression of miR-23 c,Annexin-V staining was used to observe the apoptosis of non-small cell lung cancer cells A549 under hypoxia or no hypoxia.TargetScan online analysis of the correlation between miR-23 c and ZNF420:The 3-terminal non-coding region of ZNF420 was cloned into the pmiR-RB-Repor Luciferase vector,and the dual luciferase reporter system was used to detect whether miR-23 c targets ZNF420;in the case of overexpression or knockdown of miR-23 c,by Western Blot The expression level of ZNF420 protein and the expression level of apoptosis marker protein were examined.Results When overexpressing miR-23 c,the apoptosis level of non-small cell lung cancer cells A549 was significantly increased in hypoxic environment(P<0.05).miR-23 c targets the 67-74 position of the 3′UTR of ZNF420.When overexpressing miR-23 c,the expression of ZNF420 was significantly decreased(P<0.05),the expression of cleaved-caspase9 and BAX was increased(P<0.05),and the expression of Bcl-2 was decreased(P<0.05).When miR-23 c was knocked down,the expression of ZNF420 increased significantly(P<0.05),the expression of cleavedcaspase9 and BAX decreased(P<0.05),and the expression of Bcl-2 increased(P<0.05).Conclusion miR-23 c promotes apoptosis of non-small cell lung cancer cell line A549 by targeting ZNF420 mRNA and regulating the expression of ZNF420 under hypoxia.