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目的:观察结肠癌细胞株SW480中CD133+和CD133-细胞的肿瘤干细胞样生物学特性,并分析CD133+和CD133-细胞中Bmi-1基因的表达量的差异。方法:采用流式细胞仪分选出CD133+和CD133-细胞,观察对比CD133+和CD133-细胞体外成球能力及增殖能力。用Real-time PCR检测CD133+和CD133-细胞中Bmi-1基因的表达量。结果:(1)SW480细胞中CD133+细胞所占比例为7.02%。(2)CD133+细胞体外无血清培养4 d后可成球且速度生长;CD133-细胞在干细胞培养基中不能形成细胞球,但在含血清的培养基中能贴壁分化,连续传代。无血清培养基中,CD133+细胞增殖能力强于CD133-细胞。(3)Bmi-1 m RNA在CD133+细胞的中表达显著高于CD133-细胞。结论:结肠癌细胞株SW480中CD133+所占比例很少,但其增殖能力强,Bmi-1 m RNA在CD133+细胞的中高表达,可能与肿瘤的发生发展有关。
OBJECTIVE: To observe the tumor stem cell-like biological characteristics of CD133 + and CD133- cells in human colon cancer cell line SW480 and analyze the difference of Bmi-1 gene expression in CD133 + and CD133- cells. Methods: CD133 + and CD133- cells were sorted by flow cytometry. The ability of ball formation and proliferation of CD133 + and CD133-cells in vitro were compared. Real-time PCR was used to detect the expression of Bmi-1 gene in CD133 + and CD133- cells. Results: (1) The proportion of CD133 + cells in SW480 cells was 7.02%. (2) CD133 + cells could be spheroidized and grew at high speed after 4 days of culture in vitro. CD133- cells could not form cytoplasm in stem cell culture medium, but adhered to wall in medium containing serum. In serum-free medium, CD133 + cells proliferated more strongly than CD133- cells. (3) The expression of Bmi-1 m RNA in CD133 + cells was significantly higher than that of CD133- cells. CONCLUSIONS: The proportion of CD133 + in colon cancer cell line SW480 is small but its proliferative ability is strong. The high expression of Bmi-1 m RNA in CD133 + cells may be related to tumorigenesis and development.