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目的探讨干细胞标志物三磷酸腺苷结合盒超家族G家族第2个成员(ATP-binding cassette super-family G member 2,ABCG2)在人肝癌细胞株中的表达及其与肝癌耐药的关系,观察人肝癌细胞株中ABCG2阳性表达和阴性表达细胞的功能差异。方法流式细胞仪检测ABCG2在人肝癌细胞株PLC/PRF/5、7402、7701及7721中的阳性表达,计算5-氟尿嘧啶(5-FU)、阿霉素的半数抑制浓度(IC50)值。采用免疫荧光染色观察ABCG2在肝癌7721细胞株中的表达,并进行无菌流式分选,观察ABCG2阳性表达与ABCG2阴性表达的肝癌7721细胞的功能差异。结果在4种肝癌细胞株中7721细胞株ABCG2表达阳性率最高(P<0.05),且流式细胞图中阳性表达和阴性表达细胞呈明显双峰。5-FU、阿霉素对7721细胞株的IC50值明显高于其他3种细胞株,除与5-FU对7701细胞的IC50值差异无统计学意义(P>0.05)外,其余比较差异均有统计学意义(P<0.05)。分选后的ABCG2阳性表达细胞与阴性表达细胞的增殖差异无统计学意义(P>0.05);ABCG2阳性表达细胞较阴性表达细胞更多处于相对静止期(P<0.05)。结论 ABCG2在人多种肝癌细胞株中均表达,ABCG2表达阳性率越高的细胞较表达阳性率越低的细胞更具有耐药性,ABCG2阳性表达细胞处于相对静止期。
Objective To investigate the expression of stem cell marker ATP-binding cassette super-family G member 2 (ABCG2) in human hepatocellular carcinoma cell lines and its relationship with drug resistance in hepatocellular carcinoma Functional differences of ABCG2-positive and negative-expressing cells in cell lines. Methods The positive expression of ABCG2 in human hepatocellular carcinoma cell lines PLC / PRF / 5,7402,7701 and 7721 was detected by flow cytometry. The median inhibitory concentration (IC50) of 5-fluorouracil and doxorubicin were calculated. Immunofluorescence staining was used to observe the expression of ABCG2 in hepatocellular carcinoma cell line 7721 and sterile flow cytometry. The differences between ABCG2 positive cells and ABCG2 negative hepatocellular carcinoma cells 7721 were observed. Results The positive rate of ABCG2 was the highest in 7721 cell lines (P <0.05), and the positive and negative cells in flow cytometry showed double peaks. IC50 values of 5-FU and doxorubicin on 7721 cell line were significantly higher than those of the other 3 cell lines except IC50 of 5-FU and 7701 cells (P> 0.05) There was statistical significance (P <0.05). There was no significant difference in the proliferation of ABCG2 positive cells and negative cells after sorting (P> 0.05). ABCG2 positive cells were more in quiescent than negative cells (P <0.05). Conclusion ABCG2 is expressed in many human hepatocellular carcinoma cell lines. The cells with higher positive expression rate of ABCG2 are more resistant than the cells with lower positive expression rate, and ABCG2 positive cells are in relatively quiescent stage.