上调 miR －200c 表达对人肝癌干细胞化疗耐药性的影响

摘要

目的：观察人肝癌干细胞中miR－200 c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－200 c mimics到肝癌干细胞；采用实时荧光定量RT－PCR对转染miR－200 c 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－200 c对肝癌干细胞药物敏感性的影响。结果肝癌干细胞中miR－200 c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 －200 c mimics后5－Fu和 ADM对肝癌干细胞的IC50分别为126．3 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 -200c up-regulation on the characteristics of chemotherapy re-sistance 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-200c 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-200c 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 be-fore 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-200c expres-sion in liver cancer stem cells can enhance the chemotherapy sensitivity on liver cancer stem cells ;it has the effect of reversing the chemotherapy resistance .