CAJ | 학술논문

目的动态观察人胃癌SGC-7901细胞的克隆在形态上的差异,比较不同克隆细胞的裸鼠体内成瘤能力,并鉴定含胃癌干细胞的克隆.方法采用克隆形成实验观察胃癌SGC-7901细胞克隆的形态并分类,计算克隆形成率和每种克隆的比例.采用免疫荧光技术和Western blot法检测不同克隆细胞中CD44和CDX2的表达.分离不同形态的克隆,检测其自我更新能力.将不同形态的克隆细胞分别接种于裸鼠皮下,观察各组克隆细胞的体内成瘤能力.结果 SGC-7901细胞可形成3种克隆形态,即紧密型克隆、中间型克隆和松散型克隆.SGC-7901细胞总的克隆形成率为(9.80±1.07)％,其中紧密型、中间型和松散型克隆所占的比例分别为10.2％、56.0％和33.8％.免疫荧光法检测的结果显示,CD44在紧密型克隆细胞中的表达明显强于中间型和松散型克隆,而CDX2在紧密型克隆细胞中的表达最弱.Western blot法检测结果与免疫荧光法检测的结果一致.紧密型克隆细胞具有很强的自我更新能力,在裸鼠体内的成瘤能力也较强,接种103和104个细胞的成瘤率分别为75.0％和100％.结论 SGC-7901细胞的克隆在形态和分化程度上存在差异,其中紧密型克隆可能富含胃癌干细胞.
목적 동태관찰인위암SGC-7901세포적극륭재형태상적차이,비교불동극륭세포적라서체내성류능력,병감정함위암간세포적극륭.방법 채용극륭형성실험관찰위암SGC-7901세포극륭적형태병분류,계산극륭형성솔화매충극륭적비례.채용면역형광기술화Western blot법검측불동극륭세포중CD44화CDX2적표체.분리불동형태적극륭,검측기자아경신능력.장불동형태적극륭세포분별접충우라서피하,관찰각조극륭세포적체내성류능력.결과 SGC-7901세포가형성3충극륭형태,즉긴밀형극륭、중간형극륭화송산형극륭.SGC-7901세포총적극륭형성솔위(9.80±1.07)％,기중긴밀형、중간형화송산형극륭소점적비례분별위10.2％、56.0％화33.8％.면역형광법검측적결과현시,CD44재긴밀형극륭세포중적표체명현강우중간형화송산형극륭,이CDX2재긴밀형극륭세포중적표체최약.Western blot법검측결과여면역형광법검측적결과일치.긴밀형극륭세포구유흔강적자아경신능력,재라서체내적성류능력야교강,접충103화104개세포적성류솔분별위75.0％화100％.결론 SGC-7901세포적극륭재형태화분화정도상존재차이,기중긴밀형극륭가능부함위암간세포.
Objective To dynamically investigate the morphology of human gastric cancer SGC-7901 cell clones,and then compare the tumorigenic ability of different clones in order to identify the tumor stem cell clones.Methods Clones derived from gastric cancer SGC-7901 cells were assessed by morphological observation,and the clone formation rate and proportion of each clone were calculated.The expression of CD44 and CDX2 in different clones was detected by immunofluorescence microscopy and Western blot.Furthermore,different clones were isolated and cultured,and their self-renewal property was assayed.Cells of different clones were subcutaneously inoculated into nude mice and the tumorigenic ability of each group was determined.Results Clones derived from gastric cancer SGC-7901 cells had three types,i.e.clones of tight,transitional and loose types.The total clone formation rate was (9.80 ± 1.07) ％,and the proportion of tight,transitional and loose type clones was 10.2％,56.0％ and 33.8％,respectively.The results of immunofluorescence microscopic examination showed that the signal of CD44 was significantly stronger in the tight clones than in the transitional and loose clones,however,the signal of CDX2 was weakest in the tight colonies.The results of Western blot were consistent with that of immunofluorescence microscopic observation.SGC-7901 cells of tight clones possessed strong ability of self-renewal and in vivo tumorigenicity in the nude mice.Conclusion SGC-7901 cell clones vary in morphology and differentiation,and the tight type clones may include rich gastric cancer stem cells.