【摘要】 微小RNA(microRNA,miR)在多种肿瘤细胞中均表现出下调表达,其影响肿瘤细胞的生长、迁移和侵袭能力,很可能是肿瘤生长的抑制因子,然而对于miRR-490-3p在肿瘤中的具体作用还有待进一步的阐明。本研究为了探讨miR-490-3p对人(Homo sapiens)子宫颈癌细胞HeLa生长相关特性的调控,研究了miR-490-3p对HeLa细胞生长、迁移、细胞周期及上皮间质转换(epithelial to mesenchymal transition,EMT)的影响,本研究外源感染miR-490-3p,在HeLa细胞中过表达miR-490-3p,利用qRT-PCR检测转染miR-490-3p mimic实验组与转染miR-NC mimic对照组中miR-490-3p的表达,利用细胞计数测定生长曲线,划痕实验测定迁移速度,通过流式细胞仪测定细胞周期以及Western blot检测EMT标志蛋白。结果显示,相较于对照组,实验组miR-490-3p的表达量极显著上调(P<0.01),miR-490-3p对细胞生长和迁移有抑制作用,使HeLa细胞阻滞在G2/M期,同时能抑制肿瘤细胞EMT过程。miR-490-3p抑制HeLa细胞的增殖和迁移,起着抑癌基因的作用,可以作为肿瘤的潜在治疗靶点和分子标记物,在肿瘤治疗和临床上具有重要的实践意义。更多还原

【Abstract】 microRNA-490-3p(miR-490-3p) is down-regulated and may act as a tumor inhibitor in many tumors,such as gastric cancer,ovarian carcinoma,endometrial carcinoma and so on.However,specific roles of miR-490-3p are not well understood in cancers.This study aims to investigate the effects of miR-490-3p on the proliferation and metastasis of human(Homo sapiens) cervical tumor cells HeLa.In order to investigate the effects of miR-490-3p on proliferation,migration,cell cycle and EMT(epithelial to mesenchymal transition)of HeLa cells,miR-NC mimic or miR-490-3p mimic was transfected into HeLa cells.The expression level of miR-490-3p was detected using qRT-PCR,then,cell counting was used to measure the cell proliferation,and wound healing assay was applied to detect the ability of migration.Moreover,cell cycle was assayed by flow cytometry,and EMT proteins were detected by Western blot,including E-cadherin,β-catenin,vimentin and glycogen synthase kinase 3β(GSK-3β).The results showed that,in comparison with negative controls,the expression level of miR-490-3p was significantly improved by miR-490-3p mimic transfection(P<0.01),which showed that miR-490-3p mimic was effctive to express miR-490-3p.Then,the HeLa cells transfected with miR-490-3p mimic exhibited signicantly slower growth than negative controls(P<0.05),which indicated that the overexpression of miR-490-3p reduced cell proliferation.Wound healing assay showed that,after transfected with miR-490-3p mimic,HeLa cells migration was reduced(P<0.01),which suggested miR-490-3p transfection inhibited migration ability of HeLa cells.Flow cytometry analysis confirmed that,comparing with control group,the proportion of HeLa cells transfected with miR-490-3p in G2/M phase displayed a significant increase(P<0.01).This induced cell cycle arresting at G2/M phase.miR-490-3p probably inhibited cells growth through arresting cell cycle of HeLa cells.Western blot showed that the EMT proteins were also regulated through transfection with miR-490-3p into HeLa cells,the introduction of exogenous miR-490-3p was followed by an increase in expression of E-cadherin(P<0.05) which was a biomarker of epithelial cells and decreases in β-catenin(P<0.01),vimentin(P<0.05) and GSK-3β(P<0.01) expression.The result showed that the EMT of HeLa cells was inhibited and indicated that miR-490-3p may inhibit cell invasiveness by repressing EMT,which was a key mechanism in cancer cell invasiveness and metastasis.Collectively,these results displayed the overexpression of miR-490-3p reduced cell proliferation and migration and promoted G2/M phase arrest,which suggested that miR-490-3p may act as a tumor suppressor gene in HeLa cells.These data indicates that miR-490-3p can be used as a potential therapeutic target and a molecular marker for cancer prediction in human cervical cancer.更多还原