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[摘要]
目的 探讨绿原酸对结肠癌HCT116细胞抑制作用的机制。方法 以绿原酸处理HCT116细胞,未处理组为对照组,通过基因表达谱芯片筛选出处理前后的差异表达基因,应用Real-time PCR技术对胰岛素样生长因子结合蛋白3(IGFBP3)、肺腺癌转移相关转录本1(MALAT1)、雄性性别决定基因相关高迁移率族盒基因(SOX4)、N-myc下游调节基因1(NDRG1) 4个基因进行验证,Western blotting法检测上调基因中NDRG1的蛋白表达水平。结果 基因表达谱芯片检测表明,经绿原酸处理后的结肠癌细胞中表达上调的基因为161个(差异倍数大于2),表达下调的基因为64个(差异倍数小于0.5)。这些基因主要涉及细胞信号转导、生物过程、细胞组分等功能。Real-time PCR检测证实IGFBP3、NDRG1基因经绿原酸处理后表达明显上调(P < 0.05)。Western blotting检测表明,经绿原酸处理后NDRG1基因的蛋白表达水平上调。结论 基因表达谱芯片结合Real-time PCR技术筛选出绿原酸处理后的差异表达基因,为揭示绿原酸抑制结肠癌细胞的作用机制提供依据。
[Key word]
[Abstract]
Objective To discuss the inhibition mechanism of chlorogenic acid for colorectal cancer cell line HCT116. Methods Colorectal cancer cell line HCT116 were treated with chlorogenic acid, and the untreated group was blank control. Then, the gene expression chip was used to screen the differential expression gene before and after processing, the Real-time PCR technique was used to identify IGFBP3, MALAT1, SOX4, and NDRG1, and the Western blotting detected the level of protein expression of NDRG1, which belongs to up-regulated gene. Results The gene expression spectra chip test showed that there were 161 up-regulated expression gene of colorectal cancer (the fold change was larger than 2), and 64 down-regulated expression genes (the fold change was less than 0.5) after chlorogenic acid treatment. Also, these genes were mainly related to the function of cell signal transduction, biological process and cellular component. The results of Real time PCR showed that the mRNA expression of IGFBP3 and NDRG1 were up-regulated significantly (P < 0.05) after chlorogenic acid treatment. Western Blotting results showed that the NDRG1 gene protein expression was up-regulated significantly (P < 0.05) after chlorogenic acid treatment. Conclusion The gene expression spectrum chip combined the Real-time PCR technology, which were used to screen the differential expression gene before and after the chlorogenic acid treatment, in order to reveal the inhibition mechanism of chlorogenic acid for colorectal cancer cells.
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[基金项目]
“十二五”国家科技支撑计划项目资助(2012BAD21B05)