紫檀芪对卵巢癌SKOV3细胞凋亡和糖酵解的影响及机制研究

Effect of pterostilbene on apoptosis and glycolysis of ovarian cancer SKOV3 cells and its mechanism

目的 探讨紫檀芪对卵巢癌SKOV3细胞凋亡和糖酵解的影响及机制研究。方法 分别采用0、25、50、100、150 μmol/L的紫檀芪处理SKOV3细胞24、48、72 h,用CCK-8检测紫檀芪对SKOV3细胞增殖的影响,根据CCK-8结果选择后续实验组紫檀芪浓度;采用流式细胞术检测紫檀芪对细胞凋亡的影响;采用葡萄糖氧化酶法和化学比色法检测紫檀芪对SKOV3细胞葡萄糖消耗和乳酸生成的影响;Western blot法检测信号转导与转录激活因子3(STAT3)、磷酸化的STAT3(p-STAT3)、己糖激酶2(HK2)蛋白的表达;qRT-PCR法检测葡萄糖转运蛋白1(GLUT1)、M2型丙酮酸激酶(PKM2)mRNA的表达。结果 CCK-8结果显示,紫檀芪对SKOV3细胞的增殖有抑制作用,且呈时间剂量依赖性,根据CCK-8结果,选择100 μmol/L紫檀芪处理组作为后续实验组,0 μmol/L为对照组;流式细胞术结果显示,紫檀芪可明显促进SKOV3的凋亡,浓度越大,凋亡作用越明显,差异有统计学意义(P<0.05)。此外,100 μmol/L组紫檀芪作用SKOV3细胞后,葡萄糖消耗和乳酸生成水平均较0 μmol/L组降低(P<0.01)。Western blot结果显示,与0 μmol/L组相比,100 μmol/L组p-STAT3、HK2蛋白的表达明显降低(P<0.001)。qRT-PCR结果显示,100 μmol/L组GLUT1、PKM2 mRNA的表达水平也较0 μmol/L组降低(P<0.01)。结论 紫檀芪可抑制SKOV3细胞的增殖,促进凋亡,并可能通过STAT3/HK2途径抑制卵巢癌的糖酵解。

Objective The aim of this study was to investigate the effect and mechanism of pterostilbene on apoptosis and glycolysis of ovarian cancer SKOV3 cells.Methods SKOV3 cells were treated with 0,25,50,100 and 150 μmol/L of pterostilbene for 24,48 and 72 hours.The proliferation of SKOV3 cells was measured by CCK.The effect of pterostilbene on apoptosis of SKOV3 cells was determined by flow cytometry.The glucose consumption and lactate production were detected by glucose oxidase assay and chemical colorimetry.The expression of signal transducer and activator of transcription 3(STAT3),phosphorylated STAT3(p-STAT3)and hexokinase 2(HK2)proteins was detected by Western blot.The expression of glucose transporter 1(GLUT1)and M2 pyruvate kinase(PKM2)mRNA was detected by qRT-PCR.Results Pterostilbene inhibited the proliferation of SKOV3 cells in a time- and dose-dependent manner.According to CCK-8 results,100 μmol/L of pterostilbene was selected as the follow-up experimental group and 0 μmol/L as a control group.Pterostilbene could significantly promote the apoptosis of SKOV3 cells in a dose-dependent manner.The higher the concentration,the more obvious apoptosis effect,the difference was statistically significant(P<0.05).In addition,the levels of glucose consumption and lactate production in the 100 μmol/L group were significantly lower than those in the 0 μmol/L group(P<0.01).The expression of p-STAT3 and HK2 protein in the 100 μmol/L group was also significantly lower than those in the 0 μmol/L group(P<0.001).The expression of GLUT1and PKM2 mRNA in the 100 μmol/L group was also significantly decreased than those in the 0 μmol/L group(P<0.01).Conclusion Pterostilbene can inhibit the proliferation of SKOV3 cells and promote apoptosis,and may inhibit the glycolysis of ovarian cancer through a STAT3/HK2 pathway.