南方医科大学学报2012，Vol.32Issue(3)：312-316,5.DOI:44-1627/R.20120307.1708.024
稳定表达MACC1的胃癌BGC-823/pBaBb-puro-MACC1细胞株的建立及其肿瘤相关基因的研究基因
Establishment of BGC-823/pBaBb-puro-MACC1 gastric cancer cell line stably expressing MACC1 and its tumor-related gene expression profiles
摘要
Abstract
Objective To establish a gastric cancer cell line with stable expression of metastasis-associated in colon cancer 1 (MACC1) and detect the changes in tumor-related gene expression profiles for investigating the possible regulation mechanisms between MACC1 and the differentially expressed genes. Methods The full-length MACC1 cDNA was amplified from human embryonic kidney 293FT cells and cloned into the pBaBb-puro vector. The recombinant pBaBb-puro-MACCl expression vector, after identification with restriction enzyme digestion, was transfected into 293FT cells, and the expression of fluorescent reporter gene was observed. pBaBb-puro-MACCl vector was transfected into human gastric cancer BGC-823 cell line to establish BGC-823/pBaBb-puro-MACCl cell line stably expressing MACC1. Quantitative RT-PCR and Western blotting were used to detect MACC1 expression in both BGC-823/pBaBb-puro-MACCl and control BGC-823 cells. High-throughout cDNA microarray was used to screen the effects of MACC1 on the gene expression profiles of gastric cancer cells. Results The recombinant pBaBb-puro-MACCl plasmid was successfully constructed and verified by PCR and sequencing. BGC-823/ pBaBb-puro-MACCl cells showed significantly increased MACC1 mRNA expression as compared with the control cells. The results of cDNA microarray identified 33 up-regulated and 24 down-regulated genes in the cells after MACC1 transfection involved were in various cellular functions. Conclusion The established BGC-823/pBaBb-puro-MACCl gastric cancer cell line show some important molecular changes caused by MACC1.关键词
MACC1/293FT细胞/载体构建/基因芯片Key words
metastasis-associated in colon cancer 1/ 293FT cells/ vector construction/ cDNA microarray分类
医药卫生引用本文复制引用
王妮娜,谢剑明,郑大勇,左强,廖旺军..稳定表达MACC1的胃癌BGC-823/pBaBb-puro-MACC1细胞株的建立及其肿瘤相关基因的研究基因[J].南方医科大学学报,2012,32(3):312-316,5.基金项目
国家重大科学学研究计划(2012CB945100) （2012CB945100）
广东省科技计划项目(010B031600093) （010B031600093）
