低氧耐性胃癌细胞系的建立及生物学特征分析

Establishment of hypoxia-resistant gastric cancer cell line and biological characteristics identification

目的建立低氧耐性胃癌细胞系,并分析其生物学特征,为胃癌低氧微环境研究提供良好的模型。方法通过逐渐降低氧气浓度建立耐受2%O_2的低氧耐性胃癌细胞系MGC803/hypo和BGC823/hypo。采用倒置显微镜观察细胞形态;采用Transwell小室检测细胞迁移能力;采用MTT法检测细胞增殖能力;采用免疫印迹法检测其蛋白表达;比较其与亲本胃癌细胞系的生物学特征差异。结果与亲本胃癌细胞系比较,低氧耐性胃癌细胞系呈梭形改变,且排列松散,培养液颜色易变黄,迁移能力明显增强(P <0.05),增殖能力减弱(P> 0.05);低氧相关指标HIF-2α明显上调,糖酵解相关指标Glut1显著上调,上皮间充质转化(EMT)相关指标ZO-1下调,vimentin上调;转移相关指标MUC1、integrinα5、integrinβ1上调。结论本研究建立了可耐受2%O_2浓度的低氧耐性胃癌细胞系MGC803/hypo和BGC823/hypo,其转移能力明显增强,增殖能力相对减弱,这为探索低氧微环境促进肿瘤转移机制建立了良好的模型。

Objective To establish a hypoxic-resistant gastric cancer cell line and analyze its biological characteristics to provide a good model for the study of hypoxic microenvironment in gastric cancer.Methods The hypoxic-resistant gastric cancer cell lines MGC803/hypo and BGC823/hypo,which were tolerant to 2%O2,were established by gradually decreasing the oxygen concentration.The morphology of the cells was observed by inverted microscope.The cell metastasis ability was detected by Transwell chamber.The cell proliferation ability was detected by MTT assay.The protein expression was detected by Western blot.The biological characteristics of the gastric cancer cell lines were compared with those of their parents.Results Compared with the parental gastric cancer cell line,the hypoxic-resistant gastric cancer cell line showed spindle-shaped changes and loosely arranged.The color of the culture medium was easy to turn yellow,the migration ability was significantly enhanced(P<0.05),and the proliferation ability was weakened(P>0.05).Hypoxia-related index HIF-2αwas significantly up-regulated,Glut1 was up-regulated in glycolytic index,ZO-1 was down-regulated in epithelial mesenchymal transition(EMT),vimentin was up-regulated,and metastasis-related indicators MUC1,integrinα5,and integrinβ1 were up-regulated.Conclusion This study established hypoxic-resistant gastric cancer cell lines MGC803/hypo and BGC823/hypo which can tolerate 2%O2 concentration.The Metastasis ability is obviously enhanced and the proliferative ability is relatively weakened.This provides a good model for exploring the mechanism of hypoxic microenvironment promoting tumor metastasis.