摘要
目的探讨miR-3200-3p对肝癌细胞迁移和侵袭的影响及其可能的分子机制。方法采用RT-qPCR方法检测miR-3200-3p在正常肝细胞系HL-7702与肝癌细胞系Bel-7402中的表达;构建miR-3200-3p的慢病毒低表达载体,转染Bel-7402细胞,倒置荧光显微镜及RT-qPCR检测敲低效果;根据转染病毒质粒的不同将Bel-7402细胞分为2组:LV-control组及LV-inhibitor组。Transwell迁移和侵袭实验检测2组细胞的迁移和侵袭能力;Western blot法检测2组细胞中β-catenin、MMP2、E-cadherin蛋白的表达水平。结果与HL-7702细胞比较,Bel-7402细胞中miR-3200-3p的相对表达量显著升高(P<0.001);成功建立miR-3200-3p低表达的Bel-7402稳转细胞系;与LV-control组比较,LV-inhibitor组肝癌细胞迁移和侵袭能力均明显受到抑制(均P<0.001),β-catenin(总、胞核和胞质)和MMP2蛋白表达均显著下调,E-cadherin的表达明显上调(均P<0.05)。结论miR-3200-3p在肝癌细胞中发挥促癌作用,敲低其表达可抑制肝癌细胞的迁移和侵袭,其机制可能是通过影响Wnt/β-catenin通路的活化而下调MMP2表达,上调E-cadherin而实现的。
Objective To explore the effect of miR-3200-3p on liver cancer cell migration and invasion and its possible mechanisms.Methods The expression of miR-3200-3p in human normal liver cell line HL-7702 and human hepatocellular carcinoma cell line Bel-7402 was detected by RT-qPCR.The low-expressed miR-3200-3p lentiviral vectors(LV)were constructed and transfected into Bel-7402 cells.The transfection efficiency was verified by RT-qPCR and inverted fluorescence microscopy.The Bel-7402 cells were divided into two groups:LV-control group and LV-inhibitor group.The migration and invasion ability of cells was measured by transwell assays,and the protein expression levels ofβ-catenin,MMP2 and E-cadherin were detected by Western blotting.Results The relative expression of miR-3200-3p in Bel-7402 cells was higher than that in HL-7702 cells(P<0.001).The stably transfected Bel-7402 cell line with low miR-3200-3p expression was successfully established.Compared with LV-control group,the migration and invasion were inhibited,the total,nuclear and cytoplasmicβ-catenin expression and the total MMP2 expression were reduced,and the E-cadherin expression was increased in LV-inhibitor group(P<0.001).Conclusion miR-3200-3p plays a carcinogenic role in hepatocellular carcinoma cells.Knockdown of miR-3200-3p can inhibit the migration and invasion of hepatocellular carcinoma cells,and its mechanisms may be involved in affecting the activation of Wnt/β-catenin pathway and thereby down-regulating MMP2 expression and up-regulating E-cadherin expression.
作者
冯豆
张洪
范月莹
宋玲
谭佳杰
FENG Dou;ZHANG Hong;FAN Yue-ying;SONG Ling;TAN Jia-jie(Department of Pharmacy,People’s Hospital,Wuhan University,Wuhan 430060,China;Grade 2019,Graduate School,Wuhan University,Wuhan 430060,China)
出处
《南昌大学学报(医学版)》
2022年第2期14-19,共6页
Journal of Nanchang University:Medical Sciences
基金
湖北省自然科学基金(2015CKB757)。