Y Zhao1, B Xiao, B Chen, T Qiao, D Fan. 1. Department of Gastroenterology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.
Abstract
OBJECTIVE: To investigate the role of bax in a vincristine (VCR)-induced multidrug-resistant (MDR) human gastric cancer cell line, SGC7901/VCR, in which the Bax protein expression level was significantly lower compared with that in parent cells. METHODS: A bax eukaryotic expression vector was constructed and transfected into SGC7901/VCR cells by lipofectamine, and resistant clones were selected by G418. Western blotting detected Bax expression in transfectants. Tetrazolium blue (MTT) assay evaluated the differences in drug sensitivity and cell cycle changes of transfectants were analyzed using flowcytometry (FCM). RESULTS: The bax eukaryotic expression vector was constructed and transfected into SGC7901/VCR cells. Through G418 selection, resistant clones were obtained. Western blotting demonstrated that the expression of Bax protein was markedly increased in bax transduced cells. These cells were more sensitive to adriamycin (ADR) and VCR than mock vector transducted cells. Moreover, bax transfection enhanced ADR-induced apoptosis and VCR-induced G2/M phase arrest of SGC7901/VCR cells. CONCLUSION: Bax was involved in the MDR of SGC7901/VCR cells.
OBJECTIVE: To investigate the role of bax in a vincristine (VCR)-induced multidrug-resistant (MDR) humangastric cancer cell line, SGC7901/VCR, in which the Bax protein expression level was significantly lower compared with that in parent cells. METHODS: A bax eukaryotic expression vector was constructed and transfected into SGC7901/VCR cells by lipofectamine, and resistant clones were selected by G418. Western blotting detected Bax expression in transfectants. Tetrazolium blue (MTT) assay evaluated the differences in drug sensitivity and cell cycle changes of transfectants were analyzed using flowcytometry (FCM). RESULTS: The bax eukaryotic expression vector was constructed and transfected into SGC7901/VCR cells. Through G418 selection, resistant clones were obtained. Western blotting demonstrated that the expression of Bax protein was markedly increased in bax transduced cells. These cells were more sensitive to adriamycin (ADR) and VCR than mock vector transducted cells. Moreover, bax transfection enhanced ADR-induced apoptosis and VCR-induced G2/M phase arrest of SGC7901/VCR cells. CONCLUSION:Bax was involved in the MDR of SGC7901/VCR cells.
Authors: Da-Qiang Li; Zhi-Biao Wang; Jin Bai; Jie Zhao; Yuan Wang; Kai Hu; Yong-Hong Du Journal: World J Gastroenterol Date: 2004-06-15 Impact factor: 5.742