人胆囊癌细胞系GBC-SD中侧群细胞的耐药性研究

目的 研究人胆囊细胞系GBC-SD中侧群细胞(side population cells,SP)的耐药特性,并探讨其耐药机制.方法 利用流式细胞术分选SP、非SP细胞,采用MTT检测2种细胞亚群对5种化疗药物吉西他滨、顺铂、5-氟尿嘧啶、表阿霉素、米托蒽醌的药物敏感性;利用流式细胞术检测吉西他滨处理的人胆囊癌细胞系GBC-SD中SP细胞比例的变化,并通过RT-PCR、Western印迹检测SP、非SP细胞亚群中ABCG2 mRNA和蛋白的表达.结果 吉西他滨、顺铂、5-氟尿嘧啶、米托蒽醌以对胆囊癌细胞系GBC-SD的IC50浓度分别作用SP、非SP细胞亚群1 d后,SP细胞的增殖能力高于非SP细胞,两组之间的差异具有统计学意义(P<0.05),而表阿霉素处理水平的两组间差异无统计学意义(P>0.05);人胆囊癌细胞系GBC-SD经过吉西他滨处理3周后,SP细胞的比例显著升高(8.02%±0.13%比0.62%±0.08%,P<0.05),且SP细胞明显高表达ABCG2基因.结论 人胆囊癌细胞系GBC-SD中SP细胞具有类似干细胞的耐药特性,耐药基因ABCG2高表达是其耐药的重要机制.

Abstract：

Objective To investigate the drug resistance of side population cells in human gallbladder cancer cell line GBC-SD and explore its mechanism. Methods Drug sensitivity assays of 5chemotherapeutic agents were performed on side population cells (SP) and non-SP cells of GBC-SD.GBC-SD was cultured and then treated with the chemotherapeutic agent gemcitabine. The frequency of SP by FACS was measured. RT-PCR and Western blotting were used to detect the expression of AB-CG2 in both the SP and the corresponding non-SP subsets. Results After 1 d treatment with 4 chemotherapeutic agents (gemcitabine, cisplatin, 5-fluorouracil and mitoxantrone) in IC50 concentration to GBC-SD cell line, the reproductive ability of SP was higher than that of non-SP (P<0.05). However, statistical significance was not achieved when compared with epirubicin (P>0.05). The percentage of SP in GBC-SD treated with chemotherapeutic agent gemcitabine after 3 weeks was sharply elevated by FACS (8.02% ±0.13% vs 0.62% ±0.08%, P<0.05), and the expression of ABCG2mRNA and protein were increased in SP as compared with non-SP. Conclusion SP from human gallbladder cancer cell line GBC-SD, like stem cell, showed a heighten resistance to drugs. Increased expression of ABCG2 was largely responsible for the multi-drug resistance.

Chemotherapeutic drug resistance of side population cells derived from human gallbladder cancer cell line GBC-SD

Objective To investigate the drug resistance of side population cells in human gallbladder cancer cell line GBC-SD and explore its mechanism. Methods Drug sensitivity assays of 5chemotherapeutic agents were performed on side population cells (SP) and non-SP cells of GBC-SD.GBC-SD was cultured and then treated with the chemotherapeutic agent gemcitabine. The frequency of SP by FACS was measured. RT-PCR and Western blotting were used to detect the expression of AB-CG2 in both the SP and the corresponding non-SP subsets. Results After 1 d treatment with 4 chemotherapeutic agents (gemcitabine, cisplatin, 5-fluorouracil and mitoxantrone) in IC50 concentration to GBC-SD cell line, the reproductive ability of SP was higher than that of non-SP (P<0.05). However, statistical significance was not achieved when compared with epirubicin (P>0.05). The percentage of SP in GBC-SD treated with chemotherapeutic agent gemcitabine after 3 weeks was sharply elevated by FACS (8.02% ±0.13% vs 0.62% ±0.08%, P<0.05), and the expression of ABCG2mRNA and protein were increased in SP as compared with non-SP. Conclusion SP from human gallbladder cancer cell line GBC-SD, like stem cell, showed a heighten resistance to drugs. Increased expression of ABCG2 was largely responsible for the multi-drug resistance.