CD133^(+)原代胃癌细胞生长、迁移和侵袭能力的检测

Study on proliferation,migration,and invasion of CD133^(+)primary gastric cancer cells

目的利用免疫磁珠从人原代胃癌细胞中分选CD133^(+)胃癌细胞,研究其在体内外的增殖、迁移和侵袭能力。方法利用免疫磁珠从人原代胃癌细胞中分选CD133^(+)和CD133-原代胃癌细胞,用免疫荧光检测细胞中CD133的表达,进行细胞克隆形成实验、细胞划痕实验和Transwell侵袭检测细胞体外增殖、迁移和侵袭能力;建立CD133^(+)和CD133-原代胃癌细胞裸鼠移植瘤模型,检测其体内成瘤能力及移植瘤中CD133和Ki67表达。结果用免疫磁珠成功分选到CD133^(+)和CD133-原代胃癌细胞,与CD133-原代胃癌细胞组比较,免疫荧光证实CD133^(+)细胞中CD133高表达、体外形成的细胞克隆数明显增加(P<0.001);细胞划痕和Transwell侵袭实验结果显示,CD133^(+)和CD133-细胞的伤愈百分比分别为(76.5±1.4)%和(39.8±2.0)%,侵袭细胞数分别为(624±37)/视野和(281±31)/视野,差异均有统计学意义(P<0.001);CD133^(+)细胞裸鼠内成瘤能力明显高于CD133-组,瘤组织中CD133和Ki67的表达水平也高于CD133-细胞(P<0.01)。结论CD133^(+)人原代胃癌细胞具有更强的体外增殖,迁移和侵袭能力及体内成瘤能力。

Objective To study the proliferation,migration and invasion of CD133^(+)cells,in vitro and in vivo,sorted from human primary gastric cancer cells using immunomagnetic bead.Methods By using immunomagnetic bead separation technology,CD133^(+)and CD133-primary gastric cancer cells were sorted from human primary gastric cancer cells.The immunofluorescence assay was used to detect the expression of CD133.The cell clone formation,cell wound scratch assay,and Transwell experiments were conducted to detect cell proliferation,migration and invasion in vitro,respectively.CD133^(+)and CD133-xenograft tumor models were established in nude mice,and in vivo tumorigenesis was examined and the expressions of CD133 and Ki67 were tested.Results The CD133^(+)primary gastric cancer cells were successfully obtained via immunomagnetic bead separation.Immunofluorescence confirmed that the CD133 was highly expressed in the CD133^(+)cells.Compared with the number of CD133-cell clones of 63±14,the number of cell clones formed by CD133^(+)cells in vitro was significantly increased by 201±29(P<0.001).The cell wound scratch and Transwell invasion tests showed that the percentage of scratch area in the CD133^(+)and CD133-groups was(76.5±1.4)%and(39.8±2.0)%respectively,and the number of invasions cells was 624±37 and 281±31 per field,respectively.All differences between the CD133^(+)and CD133-groups were statistically significant(P<0.001).In vivo xenograft assay showed that the tumor-forming ability of the CD133^(+)cells in nude mice was obviously higher than that of CD133-cells,and the expressions of CD133 and Ki67 were significantly increased in the xenograft tissues formed by the CD133^(+)cells compared with the CD133-cells.Conclusion The CD133^(+)primary gastric cancer cells possess much stronger in vitro proliferation,migration,and invasion abilities as well as in vivo tumorigenic ability.