ZIC1过表达对三阴性乳腺癌细胞增殖及药物敏感性的影响

The effect of ZIC1 over-expression on the proliferation of triple negative breast cancer cell and the sensitivity of drugs

目的探讨ZIC家族成员1(Zic family member 1,ZIC1)过表达对三阴性乳腺癌细胞增殖及药物敏感性的影响。方法将转染ZIC1基因和空载体的三阴性乳腺癌细胞MDA-MB-231分别作为A组和B组,将转染ZIC1基因和空载体的三阴性乳腺癌细胞MDA-MB-468分别作为C组和D组。分别采用蛋白质印迹(Western blot)法和实时荧光定量聚合酶链反应(qRT-PCR)检测ZIC1蛋白和ZIC1 mRNA的相对表达量,噻唑蓝(MTT)法检测细胞的增殖能力及对化疗药物顺铂、环磷酰胺、多西他赛、姜黄素、斑蝥素和芹菜素的敏感性。结果A组细胞中ZIC1 mRNA和ZIC1蛋白的相对表达量均明显高于B组(P﹤0.01),C组细胞中ZIC1 mRNA和ZIC1蛋白的相对表达量均明显高于D组(P﹤0.01)。培养24、36、48、60、72 h时,A组细胞的光密度(OD)值均低于同时间点B组细胞(P﹤0.05),C组细胞的OD值均低于同时间点D组细胞(P﹤0.05)。环磷酰胺、多西他赛、芹菜素对A组细胞的半数抑制浓度(IC50)均低于B组细胞(P﹤0.05),多西他赛和芹菜素对C组细胞的IC50均低于D组细胞(P﹤0.05)。结论ZIC1基因过表达能有效抑制三阴性乳腺癌细胞MDA-MB-231和MDA-MB-468的增殖,并能增强乳腺癌细胞对环磷酰胺、多西他赛及芹菜素的药物敏感性。

Objective To investigate the effect of Zic family member 1(ZIC1)overexpression on proliferation ability of triple negative breast cancer cells and the sensitivity of drugs.Method ZIC1 gene and empty vector were transfected into triple negative breast cancer cell line MDA-MB-231 as group A and group B,ZIC1 gene and empty vector were transfected into triple negative breast cancer cell line MDA-MB-468 as group C and group D.The expression of ZIC1 protein and ZIC1 mRNA were detected by Western blot and quantitative real time-polymerase chain reaction(qRT-PCR).The changes in cell proliferation ability were detected by methylthiazolyldiphenyl tetrazolium(MTT)assay,the cells sensitivity to cisplatin,cyclophosphamide,docetaxel,curcumin,cantharidin and apigenin was analyzed.Result The expression levels of ZIC1 m RNA and ZIC1 protein in group A were significantly higher than those of group B(P<0.01),the expression levels of ZIC1 mRNA and ZIC1 protein in group C were significantly higher than those of group D(P<0.01).After 24,36,48,60,72 h culturing,optical density(OD)values of group A were lower than those in group B(P<0.05),OD values of group C were lower than those in group D(P<0.05).The median inhibitory concentration(IC50)of cyclophosphamide,docetaxel and apigenin on cells in group A was lower than that in group B(P<0.05).IC50 of docetaxel and apigenin on cells in group C was lower than that in group D(P<0.05).Conclusion Over-expression of ZIC1 in triple negative breast cancer MDA-MB-231 and MDA-MB-468 cells could effectively inhibit the cell proliferation,and increase the sensitivity of cyclophosphamide,docetaxel and apigenin on breast cancer cells.