细胞周期蛋白D1在三氧化二砷抑制人胆囊癌细胞生长中的作用

Role of cyclin D1 in inhibition of human gallbladder carcinoma cells by arsenic trioxide

目的探讨细胞周期蛋白（Cyclin）D1在三氧化二砷（As2O3）抑制人胆囊癌细胞体外生长中的作用。方法不同浓度的As2O3作用于胆囊癌细胞GBC—SD，MTT法检测细胞生长情况，流式细胞仪检测细胞周期分布。Western印迹和RT—PCR检测Cyclin D1、D2、D3和CDK4、K6的蛋白和mRNA表达。构建Cyclin D1的表达质粒并转染GBC细胞，观察Cyclin D1过表达时细胞周期的变化。结果As2O3能抑制胆囊癌细胞生长，作用呈时间、剂量一效应关系；流式细胞仪检测发现As2O3可将增殖过程中的胆囊癌细胞阻滞于G1期；Western印迹及RT—PCR检测显示Cyclin D1表达下降；过表达Cyclin D1逆转了As2O3对细胞周期的改变。结论Cyclin D1在As2O3抑制人胆囊癌细胞的生长中起重要作用。

Objective To investigate the role of the cyclin D1 in inhibition of human gallbladder carcinoma （hGBC） cells by arsenic trioxide （As2O3）. Methods After the hGBC cells were cultured with different concentrations of As2O3, the proliferative activity of cells was detected by MTT methods, the cell cycle status determined by flow cytometry as well as the protein and mRNA expression of Cyclin D1,D2,D3,CDK4 and CDK6 analyzed by Western blot and RT-PCR. hGBC cells were transiently transfected with overexpression of Cyclin D1 derived from the constructed plasmid. Meanwhile, the As2O3-induced alteration of cell cycle was detected. Results The As2O3 could inhibit the growth of cells in a time- and dose-dependent manner, make cells arrested in G1 and downregulate the expression of Cyclin D1. Moreover, Cyclin D1 overexpression played the negative role in As2O3-induced alteration of cell cycle. Conclusion Cyelin D1 plays an important role in inhibition of hGBC cells by As2O3.