姜黄素联合低浓度长春新碱抑制肝癌细胞系生长的实验研究

目的:研究姜黄素联合低浓度长春新碱对肝癌细胞系HepG2的生长抑制情况及可能机制。方法:实验设对照组(仅加培养基)、姜黄素组(20μmol/L)、长春新碱组(0.5、1、2、4μg/ml)及联合用药组(20μmol/L姜黄素+1μg/ml),各组作用HepG2细胞24 h后,采用细胞计数法测定细胞增殖情况,克隆形成实验检测各处理组的长期效应,DAPI染色测定细胞的死亡方式,线粒体膜电位检测细胞的线粒体损伤情况,流式细胞术检测细胞DNA含量及细胞周期阻滞,RT-PCR检测LRP、MDR1和P21 mRNA表达的变化。结果:与对照组和单独给药组比较,姜黄素联合长春新碱组经24 h处理后,明显抑制HepG2细胞增殖能力(P<0.05);克隆形成实验显示联合作用组明显抑制细胞集落形成能力;DAPI染色显示联合用药组凋亡细胞明显增加(P<0.01),线粒体膜电位测定显示联合用药组膜电位减低可能与凋亡率增加相关;流式细胞术测定联合用药组G2/M期阻滞显著增加(P<0.05);RT-PCR检测到LRP、MDR1 mRNA的表达降低(均P<0.05),而P21 mRNA的表达升高(P<0.05)。结论:姜黄素联合低浓度长春新碱町以显著抑制HepG2生长,为化疗新方案提供了一定依据。

Synergistic tumor suppression by curcumin and low-concentration vincristine in hepatoma cell lines

OBJECTIVE： To study the synergistic effects of tumor suppression by low-concentration vincristine plus curcumin on HepG2 cell line and possible mechanisms. METHODS： Experiments included control（only medium）, curcumin only（20 IX mol/L）, vincristine only（0.5, 1, 2 and 4 Ix g/ml） and curcumin（20 μmol/L） plus vincristine（1μg/ml）. We used cell counting to assess cell growth inhibition, colony formation assay to measure long-term effect of each treatment, DAPI staining to observe cell death type, mitochondrial membrane potential test to evaluate damage on mitochondria, flow cytometry（FCM） to analyse DNA contents and cell cycle distribution, RT-PCR to find responsible genes alterations. RESULTS： Compared with control and individual treatment, after 24 h, curcumin plus vincristine treatment could significantly suppress HepG2 cell growth in cell counting assay （P〈0.05）, which was confirmed by colony formation assay. DAPI staining indicated increased apoptosis under combined treatment （P〈0.01）. Mitochondrial membrane potential was reduced which might explain the increased apoptosis. FCM showed apparent cell arrest in G2/M phase （P〈0.05）. RT-PCR detected the decrease in expressions of LRP and MDR1 as well as the increase in P21 expression （P〈0.05）. CONCLUSION： Curcumin plus low concentration vincristine could induce growth inhibition of HepG2, providing a new option for chemotherapy.