虫草素对卵巢癌细胞中Fra-1和p53表达的影响

Effect of cordycepin on the expression of Fra-1 and p53 in ovarian cancer cells

目的探讨虫草素对卵巢癌细胞中Fra-1、p53蛋白及mRNA表达的影响。方法取对数生长期SKOV3细胞,以每孔5×10^(3)个细胞接种于96孔板中,培养24 h后将细胞分为A组、B组、C组和D组,分别加入0、10、30、60μmol·L^(-1)虫草素,培养48 h。应用细胞计数试剂盒-8检测SKOV3细胞活性,Hoechst染色检测SKOV3细胞凋亡情况,Transwell实验检测细胞侵袭能力,免疫印迹法检测SKOV3细胞中Fra-1、p53蛋白的表达,实时荧光定量反转录聚合酶链反应法检测SKOV3细胞中Fra-1、p53 mRNA的表达。结果A组、B组、C组及D组SKOV3细胞活性、细胞凋亡率、穿膜细胞数比较差异均有统计学意义(P<0.001);C组和D组SKOV3细胞活性、穿膜细胞数显著低于A组、B组(P<0.05),细胞凋亡率显著高于A组、B组(P<0.05);D组SKOV3细胞活性、穿膜细胞数显著低于C组(P<0.05),细胞凋亡率显著高于C组(P<0.05)。A组与B组SKOV3细胞中Fra-1、p53蛋白及mRNA相对表达量比较差异均无统计学意义(P>0.05);C组和D组SKOV3细胞中Fra-1蛋白及mRNA相对表达量显著低于A组、B组,p53蛋白及mRNA相对表达量显著高于A组、B组(P<0.05);D组SKOV3细胞中Fra-1蛋白及mRNA相对表达量显著低于C组,p53蛋白及mRNA相对表达量显著高于C组(P<0.05)。结论虫草素可抑制SKOV3细胞活性、降低其侵袭能力、促进其凋亡,且呈现浓度依赖性,其机制可能与升高p53表达和降低Fra-1表达有关。

Objective To investigate the effect of cordycepin on the expression of Fra-1,p53 protein and mRNA in ovarian cancer cells.Methods SKOV3 cells in logarithmic growth phase were taken and inoculated into 96 well plates with 5×10^(3) cells per well.After 24 hours of culture,the cells were divided into group A,group B,group C and group D,and the cells in the four groups were added with 0,10,30 and 60μmol·L^(-1)cordycepin respectively.After 48 hours of culture,the viability of SKOV3 cells was detected by cell counting kit-8,the apoptosis of SKOV3 cells was detected by Hoechst staining,the ability of invasions was detected by Transwell test.The expression levels of Fra-1,p53 protein were detected by Western blot,the expression levels of Fra-1,p53 mRNA were detected by real-time reverse transcription polymerase chain reaction.Results There were significant differences in activity,apoptosis rates and number of transmembrane cells of SKOV3 cell among the group A,group B,group C and group D(P<0.001).The activity and the number of transmembrane cells of SKOV3 cells in group C and group D were significantly lower than those in the group A and group B(P<0.05),and the apoptosis rate of SKOV3 cells in the group C and group D was significantly higher than that in the group A and group B(P<0.05).The cell viability and number of transmembrane cells of SKOV3 in the group D were significantly lower than those in the group C(P<0.05),and the apoptosis rate of SKOV3 cells in group D was significantly higher than that the in the group C(P<0.05).There was no significant difference in the relative expression levels of Fra-1 and p53 protein and mRNA in SKOV3 cells between group A and group B(P>0.05).The relative expression levels of Fra-1 protein and mRNA of SKOV3 cells in the group C and group D were significantly lower than those in the group A and group B,and the relative expression levels of p53 protein and mRNA were significantly higher than those in the group A and group B(P<0.05).The relative expression levels of Fra-1 protein and mRNA of SKOV3 cells in the group D were significantly lower than those in the group C,and the relative expression levels of p53 protein and mRNA in SKOV3 cells in the group D were significantly higher than those in the group C(P<0.05).Conclusion Cordycepin can inhibit the activity of SKOV3 cells,reduce its invasion ability,and promote its apoptosis in a concentration dependent manner.The mechanism may be related to increasing the expression of p53 and reducing the expression of Fra-1.