Feng Jun-an
Department of gastroenterology, People's Hospital of Zhengzhou, Zhengzhou, Henan, China.
Pak J Pharm Sci. 2016 Mar;29(2 Suppl):711-8.
After the mir-155-5p over expressed in gastric cancer cells, the expression profile chip was adopted to screen its target genes. Some of the intersection of target genes were selected based on the bioinformatics prediction, in order to study the mechanism of its function and role of research. Affymetrix eukaryotic gene expression spectrum was conducted to screen mir-155-5p regulated genetic experiment. Western blot technique was employed to detect and screen the protein expression of target genes. Mimics was transfected in BGC-823 of gastric cancer cells. Compared with mimics-nc group and mock group, the mRNA expression quantities of SMAD1, STAT1, CAB39, CXCR4 and CA9 were significantly lower. After the gastric cancer cells BGC-823 and MKN-45 had been transfected by mimics, compared with mimics-nc (MNC) group and mock (MOCK) group, it was decreased for the protein expression of SMAD1, STAT1 and CAB39 in mimics (MIMICS) group. The verification of qRT-PCR demonstrated that SMAD1, STAT1, CAB39, CXCR4 and CA9 were the predicted target genes and target proteins of mir-155-5p, the over expression of mir-155-5p could enable the decreasing of its expression level in gastric cancer cells MKN-45 and BGC-823.
在胃癌细胞中过表达mir-155-5p后,采用表达谱芯片筛选其靶基因。基于生物信息学预测选择部分靶基因的交集,以研究其功能机制和作用。进行Affymetrix真核基因表达谱筛选mir-155-5p调控的基因实验。采用蛋白质印迹技术检测和筛选靶基因的蛋白质表达。将模拟物转染至胃癌细胞BGC-823中。与模拟物阴性对照(mimics-nc)组和空白组相比,SMAD1、STAT1、CAB39、CXCR4和CA9的mRNA表达量显著降低。在胃癌细胞BGC-823和MKN-45转染模拟物后,与模拟物阴性对照(MNC)组和空白(MOCK)组相比,模拟物(MIMICS)组中SMAD1、STAT1和CAB39的蛋白质表达降低。qRT-PCR验证表明,SMAD1、STAT1、CAB39、CXCR4和CA9是mir-155-5p的预测靶基因和靶蛋白,mir-155-5p的过表达可使其在胃癌细胞MKN-45和BGC-823中的表达水平降低。