Caspase-3在苦参碱诱导鼻咽癌CNE2细胞凋亡中的作用

Apoptosis and Caspase-3 changes induced by matrine in CNE2 cells

目的:探讨苦参碱对鼻咽癌CNE2细胞生长和凋亡的影响以及Caspase-3在其中的作用。方法:将不同浓度(0、0.25、1.00、4.00g/L)的苦参碱分别作用于体外培养的CNE2细胞48h;采用MTT法检测细胞的存活与增殖,采用Annexin-Ⅴ和PI双染法测定细胞凋亡,采用Westernblot法检测细胞中Caspase-3蛋白的含量。结果:在苦参碱1.00和4.00g/L浓度作用下,CNE2细胞存活和增殖受到影响,其存活率分别为对照组的65.1%和50%,与对照组相比,差异均具有统计学意义(P均<0.05);且CNE2细胞的凋亡频率明显增加,分别达到15.78%和28.44%,与对照组相比,差异均具有统计学意义(P均<0.05)。苦参碱1.00和4.00g/L浓度组细胞的Caspase-3蛋白水平分别为1.22±0.34和1.69±0.46,均比对照组0.87±0.26明显增高(P均<0.05)。结论:苦参碱能够引起鼻咽癌CNE2细胞的增殖抑制和凋亡增加,并对Caspase途径有激活作用。

OBJECTIVE：To discuss the effects of matrine on cell proliferation and apoptosis in CNE2 cells and identify the role of Caspase-3 in the process. METHODS： CNE2 cells were exposed to matrine in vitro for 48 hours. The concentrations of matrine were 0, 0.25,1.00 and 4.00 g/L. Cell proliferation was detected by MTT assay. Apoptosis was determined by double stained assay with Annexin-Ⅴ and PI. Caspase-3 protein level was determined with western blot assay. RESULTS： After treatment with 1.00,4.00 g/L matrine, the proliferation and cell viability of CNE2 cells were inhibited. It was only 65.1 and 50% of control in each group of 1.00 and 4.00 g/L matrine, respectively. At the same time, apoptosis were increased after matrine treatment. The apoptotic percentages were 15.78% and 28.44% for 1.00 and 4.00 g/L matrine treated groups, respctively. After adjustment by β-actin protein level, the analysis of grey scale indicated that Caspase-3 level were 0.87±0.26, 1.22±0.34 and 1.69±0.46 for control, 1.00 g/L matrine and 4.00 g/L matrine groups, respctively. The level of Caspase-3 was obviously increased. CONCLUSION： Matrine could inhibit cell proliferation and enhance cell apoptosis. It could also activate the caspase pathway.