ZIC1过表达对结直肠癌HCT-116细胞增殖及对苦参碱类生物碱药物敏感性的影响

The effect of ZIC1 over-expression on the proliferation of HCT-116 cells in colorectal cancer and the sensitivity of matrine alkaloids

目的:检测ZIC1基因在结直肠癌细胞内过表达后对癌细胞增殖能力的影响,以及对苦参碱类生物碱药物敏感性的改变情况。方法:首先从五种结直肠癌细胞株中筛选出ZIC1基因表达最低的一株,对其进行慢病毒转染,使其稳定表达ZIC1基因或空载体,再利用Western blot技术和qRT-PCR技术鉴定ZIC1蛋白/mRNA表达情况。通过CCK-8法和平板克隆形成实验检测过表达ZIC1基因后细胞增殖能力的改变,并利用CCK-8法分析稳转细胞株对苦参碱类生物碱药物敏感性的改变。结果:HCT116细胞株中ZIC1 mRNA/蛋白表达均显著低于其他四株(P <0. 001);慢病毒转染后,含r LV-ZIC1-PGK-Puro的重组慢病毒转染HCT116细胞内ZIC1 mRNA/蛋白表达水平显著高于对照组(P <0. 001)。CCK-8实验结果示,ZIC1基因过表达后HCT116细胞增殖能力明显降低(P <0. 01);实验组克隆形成率明显低于对照组(P=0. 003)。CCK-8法发现ZIC1过表达后HCT116结直肠癌细胞对苦参碱及槐定碱的药物敏感性得到明显的升高,而对氧化苦参碱及槐果碱的药物敏感性无明显改变。结论:HCT116结直肠癌细胞内ZIC1基因过表达后能有效抑制癌细胞的增殖,并增敏苦参碱及槐定碱对结直肠癌HCT116细胞的药理作用。

Objective:To evaluate the effect of ZIC1 gene on the proliferation ability of cancer cells in colorectal cancer cells and changes of the sensitivity of matrine alkaloid drugs.Methods:Firstly,one cell line with lowest expression of ZIC1 was selected from the five kinds of colorectal cancer cell lines,and was infected by rLV-ZIC1-PGK-Puro or rLV-PGK-Puro followed by Western blot and qRT-PCR to assess ZIC1 protein/mRNA expression.Through CCK-8 and colony formation assay,the cell proliferation was detected.CCK-8 was also used to investigate changes of the sensitivity of matrine alkaloid.Results:The expression of ZIC1 mRNA/protein in HCT116 cell lines was significantly lower than that of the other four(P<0.001).The expression level of ZIC1 mRNA/protein in HCT116 cells with rLV-ZIC1-PGK-Puro was significantly higher than that in the control group(P<0.001).The results of CCK-8 showed that the proliferation of HCT116 cells was significantly reduced after ZIC1 gene overexpression(P<0.01).The clone formation rate of the experimental group was significantly lower than that in the control group(P=0.003).CCK-8 also found that HCT116 colorectal cancer cell with over-expressed ZIC1 had higher sensitivity of matrine and sophoridine.Conclusion:Over-expressed ZIC1 in HCT116 colorectal cancer cell can effectively inhibit the proliferation of cancer cells,and increase the sensitivity of matrine and sophoridine.