黄芪扶正汤抑制人甲状腺乳头状癌细胞的机制研究

Mechanism of Huangqi Fuzheng Decoction on Inhibiting Human Thyroid Papillary Carcinoma Cells

目的:研究黄芪扶正汤抑制人甲状腺乳头状癌细胞增殖及诱导凋亡的机制。方法:体外培养人甲状腺乳头状癌TPC-1细胞,分为对照组(加完全培养基,黄芪扶正汤药物0g/L)、不同剂量实验组(黄芪扶正汤质量浓度0.5、1.0、2.5、5、10g/L)。MTT法检测各组细胞增殖情况,IC50评价检测出最优有效浓度;流式细胞术检测对照组和最优有效浓度组的细胞周期及凋亡情况;qPCR法检测对照组和最优有效浓度组细胞Rb、CDK4 mRNA的表达情况。结果:IC50评价检测系统结果显示,实验组黄芪扶正汤抑制TPC-1增殖的最优有效浓度为6.118g/L,可引起细胞发生G0/G1期阻滞;与对照组相比,最优有效浓度组细胞凋亡明显增多(P<0.05);Rb及CDK4 mRNA表达明显降低,差异均有统计学意义(均P<0.05)。结论:黄芪扶正汤可有效下调Rb及CDK4 mRNA的表达,从而抑制人甲状腺乳头状癌TPC-1细胞增殖及诱导细胞凋亡。

Objective:To study the mechanism of Huangqifuzheng decoction on inhibiting proliferation and inducing apoptosis of human papillary thyroid carcinoma cells.Methods:Human papillary thyroid carcinoma TPC-1 cells were cultured in vitro and divided into two groups:control group(with complete medium added,Huangqifuzhengdecoction 0 g/L) and test group of different doses(Huangqifuzheng decoction 0.5,1.0,2.5,5,10 g/L).Cell proliferation was measured by MTT Assay,and the optimal effective concentration was determined by IC50;Cell cycle and apoptosis were detected by flow cytometry in control group and optimal effective concentration group;the expression of Rb and CDK4 mRNA was detected by qPCR in control group and optimal effective concentration group.Results:The results of IC50 evaluation test system showed that in the test group, the optimal effective concentration of huangqi fuzheng decoction to inhibit TPC-1 proliferation was 6.118 g/L,which could cause G0/G1 phase arrest;compared with the control group, apoptosis significantly increased in the optimal effective concentration group(P<0.05);and the expression of Rb and CDK4 mRNA significantly decreased,with statistically significant differences(all P<0.05).Conclusion:Huangqifuzheng decoction can effectively down-regulate the expression of Rb and CDK4 genes,inhibit the proliferation of TPC-1 cells and induce cell apoptosis.