表没食子儿茶素没食子酸酯联合多柔比星对肺癌A549细胞增殖与荷瘤裸鼠肿瘤生长的抑制作用

Effects of Epigallocatechin Gallate Combined with Doxorubicin on Proliferation of Lung Cancer A549 Cells and Tumor Growth of Tumor-bearing Nude Mice

目的:研究表没食子儿茶素没食子酸酯(EGCG)联合多柔比星对肺癌A549细胞增殖与对荷瘤裸鼠肿瘤生长的抑制作用。方法:将A549细胞随机均分成模型(等容生理盐水)组、EGCG(50μmol/L)组、多柔比星(10μmol/L)组、联合用药(EGCG 50μmol/L+多柔比星10μmol/L)组。给药24、48 h后采用MTT法检测细胞活力并计算细胞增殖抑制率;采用流式细胞仪检测细胞凋亡状态并计算细胞凋亡率。皮下接种A549细胞于裸鼠背部以复制荷瘤裸鼠模型。24只模型小鼠随机均分为模型(等容生理盐水)组、EGCG(30 mg/kg)组、多柔比星(10 mg/kg)组、联合用药(EGCG 30 mg/kg+多柔比星10 mg/kg)组,复制模型成功后第3、6、9、12天时ip给药。称定小鼠肿瘤质量并计算抑瘤率;HE染色以进行肿瘤组织学观察。结果:与模型组比较,给药24、48 h后,EGCG组、多柔比星组、联合用药组细胞增殖抑制率、细胞凋亡率升高;与EGCG或多柔比星组比较,联合用药组细胞增殖抑制率、细胞凋亡率升高,差异均有统计学意义(P<0.01或P<0.05)。与模型组比较,EGCG组、多柔比星组、联合用药组小鼠肿瘤质量减少,抑瘤率升高,癌细胞胞核大而不规则、核质比例变小等状况有一定改善;与EGCG组或多柔比星组比较,联合用药组小鼠肿瘤质量减少,抑瘤率升高,差异均有统计学意义(P<0.01)。结论:EGCG联合多柔比星能够有效抑制A549细胞的增殖和荷瘤裸鼠肿瘤的生长,其效果好于单用药。

OBJECTIVE：To investigate the inhibition effects of epigallocatechin gallate（EGCG） combined with doxorubicin on the proliferation of lung cancer A549 cells and the growth of the tumors in tumor-bearing nude mice. METHODS：The A549 cells were randomly divided into model group（isovolumic normal saline）,EGCG group（50 μmol/L）,doxorubicin group（10μmol/L）and drug combination group（EGCG 50 μmol/L＋doxorubicin 10 μmol/L）. MTT method was used to determine cell viability after 24 and 48 h administration,and proliferation inhibition rates of cells was calculated. The flow cytometry was employed to determine the apoptosis,and apoptosis rate of cells was calculated. The back of nude mice was subcutaneously inoculated A549 cells to copy tumor-bearing model. 24 model mice were randomly divided into model group（isovolumic normal saline）,EGCG group（30 mg/kg）,doxorubicin group（10 μmol/L）and drug combination group（EGCG 30 mg/kg ＋ doxorubicin 10 μmol/L）,and were ip given drugs 3,6,9 and 12 days after successful establishment of models. The tumors of mice were weighted and tumor inhibition rates were calculated. HE stain was conducted for histological observation of tumors. RESULTS：Compared with model group,the cell proliferation inhibition rates and apoptosis rates in EGCG group,doxorubicin group and drug combination group were increased after 24 and 48 h administration. Compared with single drug group,the cell proliferation inhibition rate and apoptosis rate in drug combination group were increased,there was statistical significant difference（P〈0.01 or P〈0.05）. Compared with model group,mass of tumors in EGCG group,doxorubicin group and drug combination group was decreased and inhibition rate was increased;it had large and irregular nuclei of cancer cells and lower nucleus-cytoplasm ratios. Compared with EGCG group or doxorubicin group,mass of tumors in drug combination group was decreased and inhibition rate was increased,there was statistical significant difference（P〈0.01）. CONCLUSIONS：EGCG combined with doxorubicin can effectively inhibit the proliferation of A549 lung cancer cells and the growth of tumors,and achieve a better effect than single drug.