miRNA-93-5p通过调控FBXW7基因参与宫颈癌进展

Involvement of miRNA-93-5p in Cervical Cancer Progression by Regulating FBXW7 Gene

目的:探究miRNA-93-5p是否可通过调控FBXW7基因影响宫颈癌进展。方法:收集20例临床宫颈癌患者组织,分组为癌旁组织、宫颈癌组织,RT-qPCR检测宫颈组织中miRNA-93-5p、FBXW7 mRNA水平,并检测人正常宫颈上皮细胞H8、宫颈癌细胞Caski、HeLa、C33A细胞miRNA-93-5p、FBXW7 mRNA水平。选择宫颈癌细胞C33A,分为对照组(正常培养)、miRNA-93-5p抑制组(转染低表达miRNA-93-5p),FBXW7激活组(转染高表达FBXW7),转染48h后收集细胞,RT-qPCR检测各组细胞中miRNA-93-5p、FBXW7 mRNA水平,Western blotting检侧C33A细胞中FBXW7蛋白水平,流式细胞术检测C33A细胞凋亡率,平板克隆检测C33A细胞克隆能力,划痕实验检测C33A细胞迁移能力,Transwell实验检测C33A细胞侵袭能力。结果:与癌旁组织相比,宫颈癌组织中miRNA-93-5p mRNA水平增加、FBXW7 mRNA水平减少(P<0.001)。与H8细胞相比,Caski、HeLa、C33A细胞中miRNA-93-5p mRNA水平增加、FBXW7 mRNA水平减少(P<0.05),后续实验中选择C33A宫颈癌细胞株进行研究。与对照组相比,miRNA-93-5p抑制组C33A细胞中miRNA-93-5p mRNA水平显著降低(P<0.001),细胞凋亡率增加(P<0.001),克隆能力下降(P<0.01),细胞迁移距离减少(P<0.01),侵袭能力下降(P<0.01),FBXW7蛋白及mRNA水平显著增加(P<0.01)。与对照组相比,FBXW7激活组C33A细胞中FBXW7 mRNA水平显著增加(P<0.01),细胞凋亡率增加(P<0.001),克隆能力下降(P<0.001),迁移距离减少(P<0.01),侵袭能力下降(P<0.01)。结论:miRNA-93-5p可通过调控FBXW7基因影响宫颈癌细胞的凋亡、增殖、迁移、侵袭水平。

Objective:To explore whether miRNA-93-5p can affect the progression of cervical cancer by regulating FBXW7 gene.Methods:Twenty clinical cervical cancer patient tissues were collected and divided into cancer-adjacent tissues and cervical cancer tissues.RT-qPCR was used to detect the levels of miRNA-93-5p and FBXW7 mRNA in cervical tissues.The miRNA-93-5p and FBXW7 mRNA levels were also measured in normal cervical epithelial cells(H8)and cervical cancer cells(Caski,HeLa,C33A).The cervical cancer cell line C33A was selected for further experiments.C33A cells were divided into the control group(normal culture),miRNA-93-5p inhibition group(transfected with low-expression miRNA-93-5p),and FBXW7 activation group(transfected with high-expression FBXW7).After 48 hours of transfection,cells were collected,and RT-qPCR was used to detect miRNA-93-5p and FBXW7 mRNA levels.Western blotting measured FBXW7 protein levels in C33A cells.Flow cytometry assessed apoptosis rates,plate colony assays examined cloning ability,scratch tests evaluated migration capacity,and Transwell assays measured invasion ability.Results:In comparison with cancer-adjacent tissues,cervical cancer tissues showed increased miRNA-93-5p mRNA levels and decreased FBXW7 mRNA levels(P<0.001).Compared with H8 cells,Caski,HeLa,and C33A cells exhibited increased miRNA-93-5p mRNA levels and decreased FBXW7 mRNA levels(P<0.05).Subsequent experiments were conducted using the C33A cervical cancer cell line.In the miRNA-93-5p inhibition group,miRNA-93-5p mRNA levels significantly decreased(P<0.001),apoptosis rates increased(P<0.001),cloning ability decreased(P<0.01),migration distance reduced(P<0.01),invasion ability decreased(P<0.01),and FBXW7 protein and mRNA levels significantly increased(P<0.01)compared with the control group.In the FBXW7 activation group,FBXW7 mRNA levels significantly increased(P<0.01),apoptosis rates increased(P<0.001),cloning ability decreased(P<0.001),migration distance reduced(P<0.01),invasion ability decreased(P<0.01)compared with the control group.Conclusion:miRNA-93-5p can affect the apoptosis,proliferation,migration and invasion of cervical cancer cells by regulating FBXW7 gene.