CAJ | 학술논문

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.
목적 검측화분선위암세포계중적측군세포(SP),병통과체외실험감정기생물학특성.방법 선택3주불동분화정도적인위암세포계MKN-28、SGC-7901、BGC-823,이Hoechst33342/전화병정(PI)형광염료쌍염,유랍파미길항대조,응용류식세포의형광격활분선법검측sP세포비례;통과세포생장곡선(CCK-8법)、평판극륭형성시험대SP세포체외증식급불대칭분렬능력등생물학특성진행감정.결과 3주위암세포중균존재함량겁소적SP세포,비례재0.8%～2.7%지간.세포생장곡선현시,인위암세포계MKN-28래원적SP세포체외증식능력명현강우non-SP세포급미경분선적MKN-28세포(P＜0.05).MKN-28래원적SP세포평판극륭형성능력(89.33±6.11)%명현강우non-SP세포(13.33±1.16)%급미경분선적MKN-28세포(42.67±6.43)%,P＜0.05.MKN-28분선획득적SP세포아군경체외배양2주후가분렬산생SP세포화non-SP세포,SP세포비례유기시적100.0%하강지1.4%.결론 위암세포계중존재비례상대은정적SP세포.위암SP세포체외증식화극륭형성능력균명현강우비측군세포,차구유불대칭분렬능력,가작위위암간세포연구적절입점.
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.