Objective: To analyze the effect of bcl-2 antisense RNA on the apoptosis of promyelocytic cell line HL-60. Methods: A plasmid pDOR-AB containing bcl-2 antisense cDNA was trans fected into HL-6O cells by lipofectin, an...Objective: To analyze the effect of bcl-2 antisense RNA on the apoptosis of promyelocytic cell line HL-60. Methods: A plasmid pDOR-AB containing bcl-2 antisense cDNA was trans fected into HL-6O cells by lipofectin, and the effect of transfection was assured by DNA and RNA dot blottingI the change of bcl-2 expression and cell cycle was tested by flow cytometry; a morphologica1 change was observed by light microscope and electron microscope; and finally the sensitivity of trans fected cells to etoposide was compared with that of non-trans fected cells by gel electrophoresis. Results: pDOR-AB was successfully trans fected into HL-6o cells and its transcript was observed; Bcl-2 was down-regulated significantly ; apoptosis peak appeared before G1 phase in flow cy-tometry analysis: apoptotic cells could be seen by electron microscope, and during DNA gel electrophoresis the DNA ladder apppeared more frequently in the group trans fected with pDOR-AB than in transfected with pDOR and untransfected groups. Conclusion: Transient expression of bcl-2 antisense RNA can promote apoptosis of HL-60 cells and bco-2 plays a key role in the apoptosis of HL-60 cells.展开更多
文摘Objective: To analyze the effect of bcl-2 antisense RNA on the apoptosis of promyelocytic cell line HL-60. Methods: A plasmid pDOR-AB containing bcl-2 antisense cDNA was trans fected into HL-6O cells by lipofectin, and the effect of transfection was assured by DNA and RNA dot blottingI the change of bcl-2 expression and cell cycle was tested by flow cytometry; a morphologica1 change was observed by light microscope and electron microscope; and finally the sensitivity of trans fected cells to etoposide was compared with that of non-trans fected cells by gel electrophoresis. Results: pDOR-AB was successfully trans fected into HL-6o cells and its transcript was observed; Bcl-2 was down-regulated significantly ; apoptosis peak appeared before G1 phase in flow cy-tometry analysis: apoptotic cells could be seen by electron microscope, and during DNA gel electrophoresis the DNA ladder apppeared more frequently in the group trans fected with pDOR-AB than in transfected with pDOR and untransfected groups. Conclusion: Transient expression of bcl-2 antisense RNA can promote apoptosis of HL-60 cells and bco-2 plays a key role in the apoptosis of HL-60 cells.