淫羊藿素对人卵巢癌SKOV3细胞增殖抑制及促凋亡作用

基于PI3K/Akt信号通路,观察人卵巢癌SKOV3细胞在不同浓度淫羊藿素(icaritin)干预下的增殖与凋亡变化过程,探讨可能存在的分子机制。研究对象为人卵巢癌细胞SKOV3,设置空白对照组和淫羊藿素不同浓度组(5,10,20μmol·L^-1),药物干预48 h,采用cell counting kit-8(CCK-8)法检测淫羊藿素对卵巢癌SKOV3细胞增殖的抑制效果;通过EdU试验检测SKOV3细胞增殖能力;采用Hoechst 33342荧光染色法观察各组SKOV3细胞凋亡形态变化,通过流式细胞术检测各组细胞周期分布状况和细胞凋亡率;实时荧光定量PCR(quantitative Real-time PCR)法检测各组细胞中PTEN,PI3K,Akt基因的表达情况;Western blot法检测PTEN,PI3K,Akt,p-Akt蛋白的表达情况。结果显示,与空白对照组比较,各浓度淫羊藿素组SKOV3细胞增殖抑制率升高(P<0.05),Edu阳性细胞率均降低(P<0.05),SKOV3细胞均呈现凋亡形态学改变,SKOV3细胞凋亡率均显著上升(P<0.05),SKOV3细胞G0/G1期细胞比例均降低(P<0.05),S期细胞比例均升高(P<0.05),SKOV3细胞中PTEN的基因和蛋白表达升高,PI3K和Akt的基因表达下降,PI3K和p-Akt的蛋白表达下降(P<0.05)。结果表明,淫羊藿素可能通过调控PI3K/Akt信号通路,抑制卵巢癌SKOV3细胞增殖、诱导细胞发生凋亡并影响细胞周期分布。

Effect of icaritin on proliferation and apoptosis of human ovarian cancer cells SKOV3

Based on the PI3K/Akt signaling pathway,this study aimed to observe the proliferation and apoptosis of ovarian cancer SKOV3 cells at different concentrations of icaritin,in order to explore the possible molecular mechanisms.The research object was ovarian cancer SKOV3 cells.The cells were divided into the control group and icaritin groups(5,10,20μmol·L^-1),and administrated with drugs for 48 hours.The cell counting kit-8(CCK-8)assay was used to detect the inhibitory effect of icaritin on the proliferation of ovarian cancer SKOV3 cells.The proliferation ability of the SKOV3 cells was detected by EdU assay.Hoechst 33342 fluorescence staining was used to observe the apoptotic morphology of SKOV3 cells in each group.The distribution of cell cycle and the apoptosis rate of each group were detected by flow cytometry.Quantitative Real-time PCR was used to detect mRNA expressions of PTEN,PI3K,Akt in each group of cells.Protein expressions of PTEN,PI3K,Akt and p-Akt were measured by Western blot.The results showed that the cell inhibition rates of icaritin groups were significantly increased compared with the control group(P<0.05).The rates of EdU-positive cells of icaritin groups were significantly decreased(P<0.05).SKOV3 cells in icaritin groups showed morphological changes of apoptosis.Apoptosis rates of icaritin groups were significantly increased(P<0.05).The proportions of cells in G0/G1 phase of icaritin groups were decreased(P<0.05),while the proportions of S phase cells were increased(P<0.05).The gene and protein expressions of PTEN in icaritin groups were elevated(P<0.05).The gene expressions of PI3K and Akt in icaritin groups were down-regulated(P<0.05).The protein expression of PI3K and p-Akt in icaritin groups were reduced(P<0.05).These results indicated that icarin may inhibit the proliferation of ovarian cancer SKOV3 cells in vitro,induce cell apoptosis and affect the cycle distribution of cells by inhibiting the PI3K/Akt signaling pathway.