醛酮还原酶家族1成员B10过表达的宫颈癌细胞株Hela增殖、周期、侵袭和迁移观察

摘要

目的 观察醛酮还原酶家族1成员B10(AKR1B10)过表达的宫颈癌细胞增殖、周期侵袭和迁移情况.方法 将Hela细胞分为观察组和对照组,观察组用慢病毒系统转染AKR1B10过表达质粒,对照组转染空质粒.采用Western blotting法检测两组细胞AKR1B10、p53、基质金属蛋白酶2(MMP-2)、波形蛋白(Vimentin)蛋白;采用MTT法、克隆形成实验观察两组细胞增殖情况,结果分别用OD570、形成细胞克隆数量表示;采用流式细胞术检测两组细胞周期分布;采用划痕实验观察两组细胞迁移情况,结果以划痕愈合百分比表示;采用Transwell小室实验观察两组细胞侵袭情况,结果以穿膜细胞数表示.结果 观察组细胞 AKR1B10蛋白相对表达量高于对照组(P <0.05).MTT法测得继续培养24、48、72 h观察组细胞OD570均低于对照组(P均<0.05),克隆形成实验观察组克隆数量少于对照组(P<0.05).与对照组相比,观察组G0~G1期细胞比例上升(P<0.05),G2~M期和S期细胞比例下降(P均<0.05).观察组划痕愈合百分比低于对照组(P <0.05).观察组穿膜细胞数低于对照组(P <0.05).观察组p53蛋白相对表达量高于对照组(P<0.05),MMP-2和Vimentin蛋白相对表达量均低于对照组(P均<0.05).结论 AKR1B10过表达可抑制宫颈癌Hela细胞增殖、侵袭和迁移,阻滞细胞周期于G0~G1期,p53、MMP-2和Vimentin途径可能在其中发挥作用.%Objective To explore the effects of aldehyde ketoreductase family 1 member B10(AKR1B10)overex-pression on the cell proliferation,cell cycle,invasion,and migration of cervical cancer cells.Methods Hela cells were divided into the observation group and control group.Cells in the observation group were transfected with AKR 1B10 overex-pression plasmid by lentivirus system,and cells in the control group were transfected with empty plasmid.The expression of AKR1B10,p53,matrix metalloproteinase 2(MMP-2),and vimentin proteins in the two groups was detected by Western blotting.MTT assay and clonal formation experiment were used to analyze the proliferation of the two groups(expressed as OD570value and the number of forming cell clones).Flow cytometry was used to detect the cell cycle distribution in the two groups.Scratch test was used to analyze the migration of the two groups(expressed as the percentage of scratch healing). Transwell chamber experiment was used to analyze the cell invasion in the two groups(expressed as the number of trans-membrane cells).Results The relative protein expression level of AKR 1B10 in the observation group was higher than that in the control group(P<0.05).MTT showed that OD570value of cells in the observation group after 24-,48-and 72-hour culture was lower than that of the control group(P<0.05).The result of clone formation experiment showed that the num-ber of clones of the observation group was less than that of the control group(P<0.05).Compared with the control group,the percentage of cells in the G 0-G1phase of the observation group increased(P<0.05)while the percentage of cells in the G2-M phase and S phase decreased(P<0.05).Scratch healing percentage of the observation group was lower than that of the control group(P<0.05).The number of transmembrane cells in the observation group was lower than that in the control group(P<0.05).The relative protein expression of p53 in the observation group was higher than that in the control group(P<0.05),while the relative protein expression of MMP-2 and vimentin was lower than that of the control group(both P<0.05).Conclusion Overexpression of AKR1B10 can inhibit the proliferation,invasion,and migration of cervical cancer Hela cells,block the cell cycle in G0phase and G1phase, and the p53, MMP-2 and vimentin pathways may play a role in this process.