CONSTRUCTION AND CHARACTERIZATION OF THE MULTIDRUG-RESISTANT GASTRIC CANCER CELL STRAINS WITH UP AND DOWN-REGULATED MGr1-Ag GENE EXPRESSION
- VernacularTitle:MGr1-Ag表达上调及下调的胃癌细胞株的建立及鉴定
- Author:
Li SUN
;
Yongquan SHI
;
Changcun GUO
- Publication Type:Journal Article
- Keywords:
multidrug resistance;
stomach neoplasms;
eukaryotic expression vector
- From:
Medical Journal of Chinese People's Liberation Army
2001;0(07):-
- CountryChina
- Language:Chinese
-
Abstract:
To study the role of a novel MDR-related gene MGr1-Ag in the multidrug resistant (MDR) gastric cancer. The whole length MGr1-Ag gene was cloned, and eukaryotic vectors carrying the full length of MGr1-Ag cDNA and its antisense expression vector were constructed. The sense vector and anti-sense were then transfected into MGC803 cells and SGC7901/VCR cells respectively by lipofectamine. The result showed that a 1.0kb fragment was successfully amplified by RT-PCR and cloned into pUCm-T vector. DNA sequencing suggested that the fragment was the properly encoded MGr1-Ag gene. Recombinant eukaryotic plasmids harboring MGr1-Ag and its antisense expression vector were also obtained by subcloning the gene into pCDNA3.1/V5-His, which was confirmed by endonuclease digestion. As confirmed by Western blot, stable cell strains with up and down-regulated MGr1-Ag expression were constructed. Those cell strains provided the basis for further study on the MGr1-Ag in MDR of gastric cancer.
