芦荟大黄素对肝癌HepG2细胞放射敏感性的影响

Effects of aloe-emodin on the radiosensitivity of HepG2 cells

目的探讨芦荟大黄素(AE)对肝癌HepG2细胞放射敏感性的影响。方法采用MTT法测定AE对HepG2细胞的IC50值,作为后续实验的药物浓度。将对数生长期HepG2细胞分为对照组、AE组、γ射线组及联合组,对照组给予常规培养液处理,AE组给予IC50浓度的AE处理,γ射线组给予8 Gyγ射线处理,联合组给予IC50浓度的AE联合8 Gyγ射线处理,每组细胞均培养48 h。采用MTT法检测细胞增殖抑制率,微核试验检测细胞DNA损伤情况,流式细胞术检测细胞凋亡,克隆形成实验观察AE对HepG2细胞的放射增敏作用,Western blotting检测p53及γ-H2AX蛋白表达水平。结果AE对HepG2细胞的IC50值为60μmol·L^(-1),以此作为后续实验的药物浓度。与对照组比较,AE组、γ射线组及联合组细胞增殖抑制率、微核率、凋亡率及p53、γ-H2AX蛋白表达水平均显著升高(P<0.05),且联合组升高趋势最为明显(P<0.05);与γ射线组比较,联合组D_(0)、Dq及SF显著降低(P<0.05),SER为(1.81±0.24)。结论AE和γ射线均对HepG2细胞的增殖有抑制作用,并且AE对HepG2细胞具有放射增敏作用。

Objective To investigate the effects of aloe-emodin(AE)on the radiosensitivity of HepG2 cells.Methods MTT assay was used to determine the 50%cell growth inhibition concentration(IC50)of AE on HepG2 cells as the concentration of AE in subsequent experiments.The logarithmic phase HepG2 cells were divided into control group,AE group,γ-ray group and the combined group.The control group was given conventional culture medium;the AE group was treated with AE of IC50 concentration;theγ-ray group was given 8 Gyγ-ray radiation;the combined group was given 8 Gy radiation combined with AE of IC50 concentration.Cells in each group were cultured for 48 h.MTT method was used to detect the cell proliferation inhibition rate;micronucleus test was used to detect the DNA damage of HepG2 cells;flow cytometry was used to detect the apoptosis of cells;clone formation experiment was used to observe the radiosensitization effects of AE on HepG2 cells;Western blotting was used to detect the expressions of p53 andγ-H2AX proteins.Results The IC50 of AE on HepG2 cells was 60μmol·L^(-1),which was used as the drug concentration in the follow-up experiment.Compared with the control group,the proliferation inhibition rate,micronucleus rate and apoptosis rate of HepG2 cells in AE group,γ-ray group and the combined group were all increased,and the expression levels of p53 andγ-H2AX in cells were raised(P<0.05).And the combined group showed the most obvious increase trend of the above indexes(P<0.05).Compared with theγ-ray group,D_(0),Dq and SF in the combined group were deceased(P<0.05),and the SER was(1.81±0.24).Conclusion Both AE andγ-ray can inhibit the proliferation of HepG2 cells,and AE can enhance the radiosensitivity of HepG2 cells.