长链非编码RNA肌动蛋白纤维相关蛋白1-反义RNA1对人绒毛膜癌细胞增殖、迁移和凋亡的影响

Effect of long noncoding RNA actin filament-associated protein 1-antisense RNA1 on the proliferation,migration and apoptosis of choriocarcinoma cells

目的探讨长链非编码RNA(LncRNA)肌动蛋白纤维相关蛋白1-反义RNA1(AFAP1-AS1)对人绒毛膜癌细胞增殖、迁移和凋亡的影响。方法将人绒毛膜癌细胞JEG-3随机分为空白对照组、空白载体组和LncRNA AFAP1-AS1过表达组。空白对照组细胞未做任何处理,空白载体组细胞转染空白载体,LncRNA AFAP1-AS1过表达组细胞转染LncRNA AFAP1-AS1过表达载体。分别采用实时荧光定量聚合酶链反应和Western blot法检测JEG-3细胞中Caspase-3、Caspase-9 mRNA和蛋白的表达,采用细胞计数试剂盒-8实验、Transwell小室实验及流式细胞术检测JEG-3细胞的增殖、迁移和凋亡情况。结果 LncRNA AFAP1-AS1过表达组JEG-3细胞中Caspase-3、Caspase-9 mRNA和蛋白相对表达量均高于空白对照组和空白载体组(P <0. 05);空白对照组和空白载体组JEG-3细胞中Caspase-3、Caspase-9 mRNA及蛋白相对表达量比较差异无统计学意义(P> 0. 05)。LncRNA AFAP1-AS1过表达组细胞增殖能力、透过分子筛的细胞数显著低于空白对照组和空白载体组,细胞凋亡率显著高于空白对照组和空白载体组(P <0. 05);空白对照组与空白载体组细胞增殖能力、透过分子筛的细胞数及细胞凋亡率比较差异均无统计学意义(P>0. 05)。结论 LncRNA AFAP1-AS1可上调人绒毛膜癌JEG-3细胞中Caspase-3、Caspase-9 mRNA及蛋白水平,对JEG-3细胞的增殖及迁移有抑制作用,同时可诱导其凋亡。

Objective To investigate the effect of long noncoding RNA(LncRNA)actin filament-associated protein 1-antisense RNA1(AFAP1-AS1)on the proliferation,migration and apoptosis of choriocarcinoma cells.Methods The choriocarcinoma cells JEG-3 were randomly divided into blank control group,blank vector group and LncRNA AFAP1-AS1 overexpression group.The cells in the blank control group were not given any treatment;the cells in the blank vector group were transfected with blank vector;the cells in LncRNA AFAP1-AS1 overexpression group were transfected with LncRNA AFAP1-AS1 overexpression vector.The expressions of Caspase-3,Caspase-9 mRNA and protein in the JEG-3 cells of the three groups were detected by real-time fluorescence quantitative polymerase chain reaction and Western blot respectively.The proliferation,migration and apoptosis of JEG-3 cells were detected by cell counting kit-8 test,Transwell chamber test and flow cytometry.Results The relative expressions of Caspase-3,Caspase-9 mRNA and protein in JEG-3 cells in the LncRNA AFAP1-AS1 overexpression group were significantly higher than those in the blank control group and blank vector group(P<0.05);there was no significant difference in the relative expression of Caspase-3,Caspase-9 mRNA and protein in JEG-3 cells between the blank control group and blank vector group(P>0.05).The proliferation ability and the number of cells passing through molecular sieve in the LncRNA AFAP1-AS1 overexpression group were significantly lower than those in the blank control group and blank vector group,and the apoptotic rate was significantly higher than that in the blank control group and blank vector group(P<0.05);there was no significant difference in the proliferation ability the number of cells passing through molecular sieve and the apoptotic rate between the blank control group and blank vector group(P>0.05).Conclusion The LncRNA AFAP1-AS1 can up-regulate the expression of Caspase-3,Caspase-9 mRNA and protein in choriocarcinoma JEG-3 cells,inhibit the proliferation,migration of choriocarcinoma cells,and induce the apoptosis of cancer cells.