miR-422a靶向激肽释放酶-4抑制宫颈癌细胞的增殖、迁移与侵袭

目的探讨miR-422a靶向激肽释放酶-4(KLK4)对宫颈癌细胞增殖、迁移和侵袭的影响。方法实时荧光定量PCR法(qPCR)检测人宫颈癌细胞HeLa、SiHa和人正常宫颈上皮细胞H8中miR-422a的表达;将宫颈癌HeLa细胞分为blank组、miR-NC组、miR-422a组、mut miR-422a组、miR-422a+pcDNA组、miR-422a+pcDNA-KLK4组。CCK-8实验和Transwell实验检测各组宫颈癌HeLa细胞增殖、迁移和侵袭的能力;TargetScan软件预测miR-422a可能调控结合的靶基因,双荧光素酶活性实验对靶向关系进行验证。Western blot检测各组细胞KLK4蛋白表达水平。结果与人正常宫颈上皮细胞H8相比,宫颈癌HeLa、SiHa细胞中miR-422a呈现低表达(P<0.01),选择宫颈癌HeLa细胞进行后续实验;与miR-NC组和blank组相比,miR-422a组培养4、6 d细胞增殖能力降低,培养6、12 h细胞迁移和侵袭数量减少(P<0.05);与miR-422a+pcDNA组相比,miR-422a+pcDNA...

miR-422a targeting Kallikrein 4 inhibits proliferation,migration and invasion ofcervical cancer cells

Objective To investigate the effects of miR-422a targeting kallikinase 4 (KLK4) on proliferation, migrationand invasion of cervical cancer cells. Methods The expression levels of miR-422a in human cervical cancer cells (HeLa,SiHa) and human normal cervical epithelial cells (H8) were quantified by quantitative real-time PCR. Cervical cancer HeLacells were divided into the blank group, the miR-NC group, the miR-422a group, the mut miR-422a group, the miR-422a+pcDNA group and the miR-422a+pcDNA-KLK4 group. CCK-8 assay and transwell assay were used to detect theproliferation, migration and invasion of HeLa cells in each group. TargetScan software was used to predict the binding targetgenes. Double luciferase activity test was used to verify the targeting relationship. Western blot assay was used to detect theKLK4 protein expression in each group. Results Compared with normal human cervical epithelial cells H8, the expressionlevels of miR-422a were low in both cervical cancer HeLa cells and SiHa cell lines (P＜0.01), and cervical cancer HeLacells were selected for the subsequent experiments. Compared with that in the miR-NC group and blank group, the OD450 ofthe miR-422a group decreased after cells were cultured for 4, 6 d, the numbers of invasion and migration cells decreased inthe miR-422a group after cells were cultured for 6 and 12 h (P＜0.05). Compared with miR-422a+pcDNA group, OD450 ofthe miR-422a+pcDNA-KLK4 group was significantly increased at 4 and 6 d after culturing, and the number of cellmigration and invasion was increased at 6 and 12 h after culturing (P＜0.01). TargetScan software predicted that there was abinding site between miR-422a and KLK4, which was verified by double luciferase reporter gene experiment. Compared with the miR-NC group, the expression level of KLK4 protein was decreased in the miR-422a group, and the expressionlevel of KLK4 protein was increased in the mut miR-422a group (P＜0.01). Compared with the miR-422a+pcDNA group,the expression level of KLK4 protein increased in the miR-422a+pc DNA-KLK4 group (P＜0.01). Conclusion miR-422acan inhibit the HeLa cell proliferation, migration and invasion by targeting the KLK4 protein expression.