泥鳅体表粘液多糖诱导SGC—7901细胞凋亡机理研究

doi:10.13386/j.issn1002-0306.2012.19.004

泥鳅体表粘液多糖诱导SGC—7901细胞凋亡机理研究

Experimental study on the apoptosis of SGC-7901 induced by surface mucus polysaccharose of loach

摘要

摘要: 探讨泥鳅体表粘液多糖（PML）对人胃癌SGC-7901细胞凋亡作用和机制。采用Sevege法对PML进行分离、纯化,用紫外分光光度法、凝胶色谱法和高效液相色谱法对其鉴定后作用于SGC-7901细胞中。采用MTT法鉴定PML对该细胞的抑制作用,光显下观察细胞形态结构变化,流式细胞术检测细胞周期及凋亡,Western Blot检测Bax、Bcl-2蛋白表达变化。结果表明,用Sevege法提取PML纯度高、成分单一。MTT法,10～270μg/mL PML处理SGC-7901细胞24、48、72h,抑制作用随PML浓度和时间增加而逐渐增强。流式细胞术检测细胞凋亡,凋亡率随PML浓度和作用时间增加而逐渐增高。Western Blot检测,细胞经10～270μg/mL PML处理24、48h后Bcl-2和Bax蛋白表达随浓度和时间增加分别增强和减少,说明PML对SGC-7901细胞凋亡有促进作用,其机理与对Bcl-2与Bax基因的调控有关。

Abstract: The apoptosis effect and mechanisms of the surface mucus polysaccharide of loach (PML) on human gastric cancer SGC-7901 cell were explored.PML was isolated and purified by Sevege, and was identified by ultraviolet spectrophotometry, gel chromatography and high performance liquid chromatography before acting the role in SGC-7901 cell.Identified the inhibitive effect of PML on SGC-7901 cell by MTT, observed changes in cell morphology using optical microscope, detected cell cycle and apoptosis by flow cytometry, and detected changes of Bax, Bcl-2 protein expression by Western Blot.The results showed that it was high purity and single composition to extract PML by Sevege.The inhibition gradually increased with the increase in concentration of PML and time when the SGC-7901 cells were treated with 10～270μg/mL of PML for 24, 48, 72h.The flow cytometry showed that the apoptosis rate gradually increased with the increase in concentration of PML and time.Western blot detection indicated that Bcl-2 and Bax protein expression enhanced and reduced respectively with the increase in concentration of PML and time after the cell processing of 10～270μg/mL of PML for 24 and 48h, which explained that PML could promote the apoptosis of SGC-7901 cells and the mechanism was related to gene regulation of Bcl-2 and Bax.