蚯蚓纤溶酶抑制胃癌细胞株MGC803增殖及诱导凋亡的作用

Effect of earthworm fibrinolysin on growth inhibition and induction of apoptosis in human gastric carcinoma cell line MGC803

目的:研究蚯蚓纤溶酶对人胃癌细胞株MGC803抑制增殖及诱导凋亡的作用,并初步探讨蚯蚓纤溶酶诱导凋亡的作用机制。方法:采用MTT法观察蚯蚓纤溶酶对MGC803细胞增殖的影响,DNA凝胶电泳技术观察凋亡细胞变化,流式细胞术观察细胞凋亡率的变化,免疫细胞化学染色法检测蚯蚓纤溶酶对MGC803细胞p53蛋白表达的影响。结果:蚯蚓纤溶酶对胃癌细胞增殖有明显抑制作用,呈剂量依赖性。蚯蚓纤溶酶4 uku.ml-1作用于胃癌细胞48 h后,DNA琼脂糖凝胶电泳观察到典型梯状条带。蚯蚓纤溶酶浓度分别为2、4、8 uku.ml-1作用于胃癌细胞48 h后均可诱导细胞凋亡,各组细胞凋亡率分别为(29.51±5.54)%、(61.63±7.12)%、(80.82±5.40)%,与对照组相比,凋亡率升高有统计学意义(P<0.01)。蚯蚓纤溶酶2、3 uku.ml-1组作用于细胞48 h后,p53蛋白表达减少,平均光强度分别为1.299±0.07、1.203±0.06,与对照组相比,差异有统计学意义(P<0.01)。结论:蚯蚓纤溶酶在体外可以抑制MGC803细胞增殖,其机制可能与蚯蚓纤溶酶抑制p53蛋白表达并诱导细胞凋亡有关。

Objective To investigate the effect of earthworm fibrinolysin on growth inhibition and induction of apoptosis in human gastric carcinoma cell line MGC803 and its mechanism. Methods Cell lines were treated with a series of concentrations of earthworm fibrinolysin. Cell proliferation was detected by MTF assay. DNA gel electrophoresis was performed and cell apoptosis rates were analyzed by flow cytometry, p53 protein expression was detected by immunocytochemistry. Results MTT assay showed that earthworm fibrinolysin inhibited the proliferation of human gastric carcinoma cell line MGC803. The growth inhibition was in a dose-dependent manner. After cells were treated with earthworm fibrinolysin（4 uku·ml^-1）for 48 h, characristic DNA ladder on DNA gel electrophoresis appeared. Earthworm fibrinolysin of 2,4 and 8 uku·ml^-1at 48 h induced the apoptosis of MGC803 cells and the apoptosis rates were（29.51±5.54） %, （61.63 ± 7. 12 ） % and （ 80.82 ± 5.40） %, respectively. Compared with control group, the apoptosis rates were significantly increased （P 〈 0.01 ）. Earthworm fibrinolysin of 2 and 3 uku·ml^-1 at 48 h decreased p53 protein expression. The light intensity was 1. 299 ± 0.07 and 1. 203 ± 0.06, respectively. Compared with control group, the light intensity was significantly decreased （P 〈 0.01 ）. Conclusion Earthworm fibrinolysin inhibits the growth of human gastric carcinoma cell line MGC803 in vitro, which may be explained by its inhibitory effect on p53 and induction of apoptesis.