甲磺酸阿帕替尼联合放疗对肝癌HepG2细胞的体外协同增敏作用

Synergistic sensitization of apatinib mesylate and radiotherapy on hepatocarcinoma cells in vitro

目的:探讨甲磺酸阿帕替尼(阿帕替尼)联合放射治疗(放疗)对肝癌HepG2细胞的体外协同抑制作用,阐明其相关抗肿瘤机制。方法:体外培养肝癌HepG2细胞,以不同浓度阿帕替尼和(或)不同剂量X射线处理后,采用MTT法检测各组细胞活性,计算各组细胞增殖抑制率和阿帕替尼20%抑制浓度(IC20),并确定后续实验的X射线照射剂量。HepG2细胞分为阿帕替尼组、放疗组和阿帕替尼+放疗组(联合组),流式细胞术检测各组细胞凋亡率,细胞划痕实验检测各组细胞迁移率,酶联免疫吸附试验(ELISA)法检测各组细胞培养上清中血管内皮生长因子(VEGF)水平。结果:MTT法检测,阿帕替尼的IC20为1.32μmol·L^(-1),以此作为后续实验阿帕替尼作用浓度,确定2 Gy为后续实验X射线照射剂量。与对照组比较,阿帕替尼组和放疗组细胞凋亡率差异无统计学意义(P>0.05),联合组细胞凋亡率升高(P<0.05)。与对照组比较,阿帕替尼组、放疗组和联合组细胞迁移率降低(P<0.05);与阿帕替尼组和放疗组比较,联合组细胞迁移率降低(P<0.05)。与对照组比较,阿帕替尼组和联合组细胞培养上清中VEGF水平降低(P<0.05);与阿帕替尼和放疗组比较,联合组细胞培养上清中VEGF水平降低(P<0.05)。结论:阿帕替尼联合放疗在体外可明显抑制肝癌HepG2细胞增殖和迁移,并诱导细胞凋亡,其作用可能与抑制细胞VEGF分泌有关。

Objective:To discuss the synergistic inhibitory effect of apatinib mesylate(apatinib)combined with radiotherapy(RT)on the hepatocellular carcinoma(HepG2)cells in vitro,and to clarify its related antitumor mechanism.Methods:The HepG2 cells were cultured in vitro and treated with different concentrations of apatinib and/or varying doses of X-rays.MTT method was used to detect the survival rates of the cells in various groups;the inhibitory rates of cell proliferation and the 20%inhibitory concentration(IC20)of apatinib were calculated;the X-ray irradiation dose for subsequent experiments was detected.The HepG2 cells were divided into apatinib group,RT group,and apatinib+RT group(combined group).Flow cytometry was used to detect the apoptotic rates of the cells in various groups;wound healing assay was used to detect the migration rates of the cells in various groups;ELISA method was used to detect the levels of vascular endothelial growth factor(VEGF)in the cell culture supernatant in various groups.Results:The MTT results showed that the IC20 of apatinib was 1.32μmol·L^(-1),and this concentration was used for subsequent experiments,and the X-ray irradiation dose for the follow-up experiments was 2 Gy.Compared with control group,the apoptotic rates of the cells in apatinib group and RT group had no significant differences(P>0.05),while the apoptotic rate of the cells in combined group was increased(P<0.05).Compared with control group,the migration rates of the cells in apatinib group,RT group,and combined group were decreased(P<0.05);compared with apatinib group and RT group,the migration rate of the cells in combined group was decreased(P<0.05).Compared with control group,the levels of VEGF in the cell culture supernatant in apatinib group and combined group were decreased(P<0.05);compared with apatinib and RT group,the level of VEGF in the cell culture supernatant in combined group was decreased(P<0.05).Conclusion:Apatinib combined with radiotherapy significantly inhibits the proliferation and migration of the HepG2 cells in vitro and induces the apoptosis;its effect may be related to the inhibition of VEGF secretion by cells.