目的探索在人胆囊癌细胞系中是否存在具有干细胞特性的SP细胞(side population cells)以及SP细胞、非SP细胞、GBCSD细胞系之间在耐药基因ABCG2和胚胎干细胞表面标志Oct-4表达方面的差异性。方法采用FACS(流式激活细胞分选)技术分选出人胆囊癌SP细胞和非SP细胞,通过RTPCR、Western blot以及流式技术来检测SP细胞、非SP细胞以及GBC—SD细胞系在ABCG2和Oct-4表达方面的差异情况。结果人胆囊癌GBC—SD细胞系中存在着具有干细胞潜能的SP细胞,其所占的比例为(0.64±0.08)%。并且ABCG2在胆囊癌SP细胞中呈现出高表达的状态[(89.56±3.86)%].在非SP细胞中几乎不表达[(1.32±0.49)%],两者之间的差异具有统计学意义(P〈0.05).在GBCSD细胞系中呈弱表达[(12.37±1.61)%]。Oct-4在三种细胞中的表达分别为:(94.87±1.40)%、(88.16±2.34)%、(90.17±1.61)%,三者之间差异没有统计学意义(P〉0.05)。结论人胆囊癌的起源与肿瘤干细胞密切相关并且在胆囊癌细胞系中存在着具有干细胞特性并且高表达耐药基因ABCG2的人胆囊癌SP细胞。
Objective To investigate whether the side population cells (SP cells) exist in human gallbladder carcinoma cell line and the difference of ABCG2 and Oct-4 gene expression in SP cells, non SP cells and GBC SD cell line. Methods Fluorescence-activated cell sorting (FACS) was used to sort the side population cells and non-side population cells from GBC-SD cell line of gallbladder carcinoma of human being. Then. the sorting cells were cultured and the expression of ABCG2 and Oct 4 detected among SP cells, non-SP cells and GBC SD cell line by using reverse transcription polymerase chain reaction (RT-PCR). flow cytometry and western blot technique. Results A very small fraction of cells [(0.64±0.08)%] were isolated through FACS analysis and highly expressed ABCG2 gene [(89.56± 3.86)%]. On the contrary, ABCG2 was nearly no expression in non-SP cells [(1.32 ± 0.49) %] and lower expression in GBC SD cell line[( 12.37 ± 1.61) % ] . Meanwhile, the expression of Oct 4 in the 3 kinds of cells was (94.87±1.40)%, (88.16±2.34)% and (90.17±1.61)%, respectively. Conclusion The origin of human gallbladder carcinoma has close relationship with tumor stem cells and the side population cells which has the potency of stem ceils highly expressing ABCG2 exit in the GBC SD cell line.