红景天苷抑制卵巢癌细胞SKOV3增殖的基因水平研究

Salidroside Inhibits Expression Level of Proliferation Gene of SKOV3 Ovarian Cancer Cells

目的:探究红景天苷对卵巢癌细胞SKOV3增殖的作用及作用机制.方法:在培养基中添加不同浓度(4、8、16、30和40μmol/mL)红景天苷,分别标示为SAL-1、SAL-2、SAL-3、SAL-4和SAL-5组,采用CCK-8检测细胞活性,流式细胞术检测SKOV3细胞的细胞周期和凋亡率,实时荧光定量PCR(RT-qPCR)和Western blot检测磷酸酶与张力蛋白同源基因(PTEN)的mRNA水平和蛋白水平的表达.结果:对照组(CG)中SKOV3细胞生长良好,增殖旺盛.与CG组相比,SKOV3细胞数量在48h和72h时相对减少,SKOV3细胞增殖的抑制率随着红景天苷剂量的增加而增大.SAL-5组的抑制率明显高于CG组和SAL-1组(P<0.05或P<0.01).SAL-5组在24h、48h和72h时,SKOV3细胞的凋亡率均显著高于CG组和SAL-1组(P<0.05或P<0.01).G0/G1期细胞比例逐渐增加,S期细胞比例逐渐减少,SKOV3细胞中的PTEN基因和蛋白表达随红景天苷浓度的增加而升高(P<0.05),SAL-5组为最大值.结论:红景天苷可能通过促进SKOV3细胞PTEN基因mRNA和蛋白的表达水平,剂量依赖性地抑制SKOV3细胞的增殖活力,影响细胞分裂过程,高剂量的红景天苷促进SKOV3细胞凋亡.

Objective:To investigate the effect and mechanism of Salidroside on proliferation of ovarian cancer cells SKOV3.Methods:Salidroside with different concentrations(4,8,16,30 and 40μmol/mL)was added to the culture medium,labeled as SAL-1,SAL-2,SAL-3,SAL-4 and SAL-5 groups,respectively.Cell activity was detected by CCK-8.Cell cycle and apoptosis rates of SKOV3 cells were detected by flow cytometry,respectively.Real-time fluorescence quantitative PCR(RT-qPCR)and Western blotting were used to detect the mRNA and protein expression of phosphatase and tensin homolog gene(PTEN).Results:SKOV3 cells in the control group(CG)grew well and proliferated vigorously.In comparison with CG group,the numbers of SKOV3 cells were decreased at 48h and 72h,and the inhibition rates of SKOV3 cell proliferation were increased with the increase of salidroside doses.The inhibitory rate of SAL-5 group was significantly higher than that of CG group and SAL-1 group(P<0.05 or P<0.01).The apoptosis rate of SKOV3 cells in SAL-5 group was significantly higher than that in CG group and SAL-1 group at 24h,48h and 72h(P<0.05 or P<0.01).The proportion of G0/G1 phase cells was gradually increased,while the proportion of S phase cells gradually decreased.The expression of PTEN gene and protein in SKOV3 cells increased with the increase of salidroside concentration(P<0.05),and the maximum value was found in SAL-5 group.Conclusion:Salidroside inhibited SKOV3 cell proliferation and affected cell division in a dose-dependent manner by promoting the expression levels of PTEN gene mRNA and protein in SKOV3 cells.High dose salidroside could promote the apoptosis of SKOV3 cells.The results of this study provided new strategies and potential therapeutic targets for the diagnosis and treatment of ovarian cancer.