氧化苦参碱对人食管癌细胞株Eca109增殖及凋亡的影响

Effects of oxymatrine on the proliferation and apoptosis of human esophageal carcinoma Eca109 cell line

目的:观察氧化苦参碱(Oxy)对食管癌细胞株Eca109的抑制增殖及诱导凋亡作用。方法:培养食管癌细胞株(Eca109),以MTT比色法、生长曲线、及流式细胞术测定Oxy对食管癌细胞株Eca109抑制增殖及诱导凋亡的作用。免疫沉淀法并ERK活性试剂盒测定ERK活性;免疫印记法测定p-ERK1/2、Cyclin D1、p21waf/cip1、Bax及Bcl-2表达。结果:MTT实验及生长曲线显示Oxy明显抑制Eca109细胞增殖,流式细胞术显示Oxy可诱导Eca109细胞凋亡;免疫沉淀法显示Oxy可抑制ERK活性,免疫印记法显示Oxy抑制p-ERK1/2、Cyclin D1及Bcl-2表达,同时上调p21waf/cip1和Bax表达,Bax/Bcl-2比值增加。结论:氧化苦参碱可以抑制食管癌细胞株Eca109增殖,机制与影响ERK及下游CyclinD1、p21waf/cip1表达有关;其诱导凋亡途径与上调Bax表达,降低Bcl-2表达有关。

OBJECTIVE To observe the effects of oxymatrine on the proliferation and apoptosis of human esophageal carci noma Eca109 cell line. METHODS The effect of oxymatrine on the proliferation of Eca109 cells was observed by MTT assay and growth curve. Detection of apoptosis was performed with flow cytometry. ERK activity was measured by immuno-Precipitation, the expressions of p-ERK1/2, Cyclin D1, p21waf/cip1, Bax and Bcl-2 were analyzed by western blot. RESULTS oxymatrine could depress the proliferation of Eca109 cell line. The intracellular ERK activity and expressions of p-ERK1/2 and Cyclin D1 were decreased, while p21waf/cip1 expression was elevated by different concentrations of Oxymatrine. Moreover, Oxymatrine could induce apoptosis of Eca109, the mechanisms were related with upregulation of Bax expression and downregu lation of Bcl-2 expression. CONCLUSION Oxymatrine can depress the proliferation of Ecal09 cell line with inhibiting activity of ERK and expression of p-ERK1/2. Meanwhile, oxymatrine can induce apoptosis of Eca109 cell line with enhancing Bax expression and depressing Bcl 2 expression.