[Effect of total saponins of Panaxginseng on bax and bcl-xl gene expression in HL-60 cells].
Xi-Min Fang, Li Yuan, Jiangchao Qian, Haixia Zhou, Juxiang Wang
Abstract
Xi-Min Fang, Li Yuan, Jiangchao Qian, Haixia Zhou, Juxiang Wang
Abstract
The aim of this study was to investigate the effect of the total saponins of Panaxginseng (TSPG) on cells expression of apoptosis-related genes bax and bcl-xl in HL-60 cells and its mechanism inducing apoptosis of HL-60 cells. The morphology of HL-60 cells was observed under normal and fluorescence microscopes; the percentage of apoptotic HL-60 cells was assayed by flow cytometry and the DNA ladder was observed by DNA agarose gel electrophoresis; the expression changes of bax and bcl-xl mRNAs were detected by RT-PCR after HL-60 cells were treated with TSPG at final concentrations of 0, 100, 200, 400, 800 and 1600 microg/ml for 48 hours. The results showed that the percentage of apoptotic HL-60 cells went up as the dose increased, the typical apoptotic cell morphology and the appearance of apoptotic DNA ladder could be observed when treated with 0 - 400 microg/ml TSPG for 48 hours. At the same time and same range of concentration, the expression of bax mRNA increased and the bcl-xl expression decreased gradually. When higher than 400 microg/ml of TSPG was used, cell necrosis appeared and the percentage of apoptotic HL-60 cells even decreased. It is concluded that the apoptosis or necrosis in HL-60 cells can be induced by TSPG at certain range of concentration, and the percentage of apoptosis is dose-dependent. The effect on up-regulation of bax mRNA and down-regulation of bcl-xl mRNA probably play an important role in apoptosis of HL-60 cells induced by TSPG.
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The aim of this study was to investigate the effect of the total saponins of Panaxginseng (TSPG) on cells expression of apoptosis-related genes bax and bcl-xl in HL-60 cells and its mechanism inducing apoptosis of HL-60 cells. The morphology of HL-60 cells was observed under normal and fluorescence microscopes; the percentage of apoptotic HL-60 cells was assayed by flow cytometry and the DNA ladder was observed by DNA agarose gel electrophoresis; the expression changes of bax and bcl-xl mRNAs were detected by RT-PCR after HL-60 cells were treated with TSPG at final concentrations of 0, 100, 200, 400, 800 and 1600 microg/ml for 48 hours. The results showed that the percentage of apoptotic HL-60 cells went up as the dose increased, the typical apoptotic cell morphology and the appearance of apoptotic DNA ladder could be observed when treated with 0 - 400 microg/ml TSPG for 48 hours. At the same time and same range of concentration, the expression of bax mRNA increased and the bcl-xl expression decreased gradually. When higher than 400 microg/ml of TSPG was used, cell necrosis appeared and the percentage of apoptotic HL-60 cells even decreased. It is concluded that the apoptosis or necrosis in HL-60 cells can be induced by TSPG at certain range of concentration, and the percentage of apoptosis is dose-dependent. The effect on up-regulation of bax mRNA and down-regulation of bcl-xl mRNA probably play an important role in apoptosis of HL-60 cells induced by TSPG.
Key concepts: Apoptosis, Molecular biology, Agarose gel electrophoresis, Bcl-xL, Flow cytometry, Biology, Gene expression, Necrosis