CONSTRUCTION AND CHARACTERIZATION OF THE MULTIDRUG-RESISTANT GASTRIC CANCER CELL STRAINS WITH UP AND DOWN-REGULATED MGr1-Ag GENE EXPRESSION / 解放军医学杂志

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.