上调miR-200c表达对人肝癌干细胞化疗耐药性的影响

Influence of miR-200c up-regulation on chemotherapy resistance of liver cancer stem cells

目的观察人肝癌干细胞中miR-200c的表达改变对化疗药物敏感性的影响。方法利用流式细胞分选技术从MHCC97人肝癌细胞系中分离出肝癌干细胞。应用脂质体介导转染miR-200c mimics到肝癌干细胞;采用实时荧光定量RT-PCR对转染miR-200c mimics前后肝癌干细胞中miR-200c表达水平进行检测;通过噻唑蓝(MTT)法检测miR-200c对肝癌干细胞药物敏感性的影响。结果肝癌干细胞中miR-200c的相对表达量为0.17±0.02,转染miR-200c mimics后肝癌干细胞中miR-200c的相对表达量为1.24±0.28,转染前后比较差异有统计学意义(P<0.05)。转染前5-氟尿嘧啶(5-Fu)和阿霉素(ADM)对肝癌干细胞的半数抑制浓度(IC50)分别为22.74 mg/L和9.56 mg/L,转染miR-200c mimics后5-Fu和ADM对肝癌干细胞的IC50分别为12.63 mg/L和3.51 mg/L,转染前后比较差异均有统计学意义(P均<0.05)。结论上调人肝癌干细胞中miR-200c的表达可以增强肝癌干细胞的化疗敏感性,具有逆转其化疗耐药的作用。

Objective It is to observe the influence of miR- 200 c up-regulation on the characteristics of chemotherapy resistance of liver cancer stem cells. Methods Liver cancer stem cells were isolated by fluorescence activated cell sorting（ FACS） from MHCC97 cell line of human hepatocellular carcinoma. The miR- 200 c mimics were transfected into liver cancer stem cells by liposome. The expression levels of miR- 200 c in liver cancer stem cells were measured by reverse transcription polymerase chain reaction（ RT- PCR）. MTT method was used to evaluate the chemotherapy resistance of liver cancer stem cells. Results The expression of miR- 200 c in liver cancer stem cells was 0. 17 ± 0. 02 before transfection,and was 1. 24 ±0. 28 after transfection,there was significant difference between before and after transfection（ P〈0.05）. The half inhibition concentration（ IC50） of 5- fluorouracil（ 5- Fu） and adriamycin（ ADM） were 22. 74 mg/L and 9. 56 mg/L respectively before transfection on liver cancer stem cells,and were 12. 63 mg / L and 3. 51 mg / L respectively after transfection,there was significant difference between before and after transfectin（ all P〈0.05）. Conclusion Up-regulation of the miR- 200 c expression in liver cancer stem cells can enhance the chemotherapy sensitivity on liver cancer stem cells; it has the effect of reversing the chemotherapy resistance.