胃癌侧群细胞分选及生物学特性的体外鉴定

目的 检测和分选胃癌细胞系中的侧群细胞(SP),并通过体外实验鉴定其生物学特性.方法 选择3株不同分化程度的人胃癌细胞系MKN-28、SGC-7901、BGC-823,以Hoechst33342/碘化丙锭(PI)荧光染料双染,维拉帕米拮抗对照,应用流式细胞仪荧光激活分选法检测sP细胞比例;通过细胞生长曲线(CCK-8法)、平板克隆形成试验对SP细胞体外增殖及不对称分裂能力等生物学特性进行鉴定.结果 3株胃癌细胞中均存在含量极少的SP细胞,比例在0.8%～2.7%之间.细胞生长曲线显示,人胃癌细胞系MKN-28来源的SP细胞体外增殖能力明显强于non-SP细胞及未经分选的MKN-28细胞(P＜0.05).MKN-28来源的SP细胞平板克隆形成能力(89.33±6.11)%明显强于non-SP细胞(13.33±1.16)%及未经分选的MKN-28细胞(42.67±6.43)%,P＜0.05.MKN-28分选获得的SP细胞亚群经体外培养2周后可分裂产生SP细胞和non-SP细胞,SP细胞比例由起始的100.0%下降至1.4%.结论 胃癌细胞系中存在比例相对稳定的SP细胞.胃癌SP细胞体外增殖和克隆形成能力均明显强于非侧群细胞,且具有不对称分裂能力,可作为胃癌干细胞研究的切入点.

Preliminary Identification of side population cells isolated from gastric cancer

Objective To identify and characterize the side population cells isolated from cell lines of gastric cancer.Methods Side population cells of three human gastric cancer ceU lines with different differentiation grades.named MKN-28,SGC-7901,and BGC-823,were analyzed and isolated by fluorescence-activated cell sorting after being stained by fluorochrome Hoechst33342 and PI,then characterized bv cell growth curve(CCK-8)and colony formation assay.Results Side population ceUs were detected in all of the three gastric cancer cell hnes,with a proportion of 0.8%-2.7%.Cell growth curve indicated that proliferative capacity of the side population ceUs isolated from MKN-28 Was better than the total cells without sorting and the non-SP cells(P＜0.05).Colony formation assay showed that the colony-forming efficiency of the side population cells was hJigher than the total ceHs without sorting and the non-SP cells (89.33% vs 42.67% vs 13.33%,P＜0.05).SP cells isolated from MKN-28 could finally generate both SP and non-SP cells through a dissymmetry fission.Conclusion Side population ceHs could be detected in the cell lines of gastric cancer.With a potential capacity of dissymmetry fission.sP eeUs showed a better proliferative ability and higher colony-forming efficiency in vitro than non-SP cells.