目的:研究促凋亡药物Navitoclax(NTX)联合化疗药物柔红霉素(Daunorubicin,DNR)对红白血病细胞凋亡的影响。方法:取对数生长期的K562、HEL和TF-1细胞,给予NTX、DNR以及2药联合,CCK-8检测细胞生长,Annexin V-DAPI双染流式细胞术检测细胞...目的:研究促凋亡药物Navitoclax(NTX)联合化疗药物柔红霉素(Daunorubicin,DNR)对红白血病细胞凋亡的影响。方法:取对数生长期的K562、HEL和TF-1细胞,给予NTX、DNR以及2药联合,CCK-8检测细胞生长,Annexin V-DAPI双染流式细胞术检测细胞的凋亡,real-time RT-PCR检测凋亡相关基因BAX,BAK,BCL-2、BCL-xl、BIM的表达,对比分析NTX、DNR与2药联合对K562、HEL和TF-1细胞凋亡的影响。结果:NTX联合DNR能明显抑制K562、HEL和TF-1细胞的生长;凋亡检测结果显示,联合用药组K562、HEL和TF-1细胞凋亡率明显高于NTX、DNR单用药组(P<0.05);凋亡相关基因检测结果显示,K562细胞前凋亡蛋白基因BAK、BAX的表达水平在联合用药组明显高于2个单药组,抗凋亡蛋白基因BCL-2、BCL-xl的表达水平明显低于2个单药组(P<0.05);HEL细胞联合用药24 h BAK的表达水平高于DNR单药组(P<0.05);TF-1细胞联合用药24 h BCL-2的表达低于2个单用药组,48 h BAK的表达联合用药组最高,BCL-2、BCL-xl的表达水平在联合用药组低于NTX单药组(P<0.05)。结论:NTX联合DNR能明显促进红白血病细胞系K562、HEL和TF-1细胞凋亡,诱导凋亡相关基因的表达。本研究期待为红白血病的临床治疗提供新方案。展开更多
Background:The combination of senescence triggers with senolytic drugs is considered a promising new approach to cancer therapy.Here,we studied the efficacy of the genotoxic agent etoposide(Eto)and irradiation in indu...Background:The combination of senescence triggers with senolytic drugs is considered a promising new approach to cancer therapy.Here,we studied the efficacy of the genotoxic agent etoposide(Eto)and irradiation in inducing senescence of Panc02 pancreatic cancer cells,and the capability of the Bcl-2 inhibitor navitoclax(ABT-263;Nav)to trigger senolysis.Methods:Panc02 cells were treated with Eto or irradiated with 5–20 Gy before exposure to Nav.Cell survival,proliferation,and senescence were assessed by trypan blue staining,quantification of DNA synthesis,and staining of senescence-associatedβ-galactosidase(SA-β-Gal)-positive cells,respectively.Levels of mRNA were determined by real-time polymerase chain reaction,and protein expression was analyzed by immunoblotting.Panc02 cells were also grown as pancreatic tumors in mice,which were subsequently treated with Eto and Nav.Results:Eto and irradiation had an antiproliferative effect on Panc02 cells that was significantly or tendentially enhanced by Nav.In vivo,Eto and Nav together,but not Eto alone,significantly reduced the proportion of proliferating cells.The expression of the senescence markerγH2AX and tumor infiltration with T-cells were not affected by the treatment.In vitro,almost all Eto-exposed cells and a significant proportion of cells irradiated with 20 Gy were SA-β-Gal-positive.Application of Nav reduced the percentage of SA-β-Gal-positive cells after irradiation but not after pretreatment with Eto.In response to triggers of senescence,cultured Panc02 cells showed increased protein levels ofγH2AX and the autophagy marker LC3B-II,and higher mRNA levels of Cdkn1a,Mdm2,and PAI-1,while the effects of Nav were variable.Conclusions:In vitro and in vivo,the combination of senescence triggers with Nav inhibited tumor cell growth more effectively than the triggers alone.Our data also provide some evidence for senolytic effects of Nav in vitro.展开更多
文摘目的:研究促凋亡药物Navitoclax(NTX)联合化疗药物柔红霉素(Daunorubicin,DNR)对红白血病细胞凋亡的影响。方法:取对数生长期的K562、HEL和TF-1细胞,给予NTX、DNR以及2药联合,CCK-8检测细胞生长,Annexin V-DAPI双染流式细胞术检测细胞的凋亡,real-time RT-PCR检测凋亡相关基因BAX,BAK,BCL-2、BCL-xl、BIM的表达,对比分析NTX、DNR与2药联合对K562、HEL和TF-1细胞凋亡的影响。结果:NTX联合DNR能明显抑制K562、HEL和TF-1细胞的生长;凋亡检测结果显示,联合用药组K562、HEL和TF-1细胞凋亡率明显高于NTX、DNR单用药组(P<0.05);凋亡相关基因检测结果显示,K562细胞前凋亡蛋白基因BAK、BAX的表达水平在联合用药组明显高于2个单药组,抗凋亡蛋白基因BCL-2、BCL-xl的表达水平明显低于2个单药组(P<0.05);HEL细胞联合用药24 h BAK的表达水平高于DNR单药组(P<0.05);TF-1细胞联合用药24 h BCL-2的表达低于2个单用药组,48 h BAK的表达联合用药组最高,BCL-2、BCL-xl的表达水平在联合用药组低于NTX单药组(P<0.05)。结论:NTX联合DNR能明显促进红白血病细胞系K562、HEL和TF-1细胞凋亡,诱导凋亡相关基因的表达。本研究期待为红白血病的临床治疗提供新方案。
基金supported by a grant from the Bundesministerium fur Bildung und Forschung(01ZX1903A).
文摘Background:The combination of senescence triggers with senolytic drugs is considered a promising new approach to cancer therapy.Here,we studied the efficacy of the genotoxic agent etoposide(Eto)and irradiation in inducing senescence of Panc02 pancreatic cancer cells,and the capability of the Bcl-2 inhibitor navitoclax(ABT-263;Nav)to trigger senolysis.Methods:Panc02 cells were treated with Eto or irradiated with 5–20 Gy before exposure to Nav.Cell survival,proliferation,and senescence were assessed by trypan blue staining,quantification of DNA synthesis,and staining of senescence-associatedβ-galactosidase(SA-β-Gal)-positive cells,respectively.Levels of mRNA were determined by real-time polymerase chain reaction,and protein expression was analyzed by immunoblotting.Panc02 cells were also grown as pancreatic tumors in mice,which were subsequently treated with Eto and Nav.Results:Eto and irradiation had an antiproliferative effect on Panc02 cells that was significantly or tendentially enhanced by Nav.In vivo,Eto and Nav together,but not Eto alone,significantly reduced the proportion of proliferating cells.The expression of the senescence markerγH2AX and tumor infiltration with T-cells were not affected by the treatment.In vitro,almost all Eto-exposed cells and a significant proportion of cells irradiated with 20 Gy were SA-β-Gal-positive.Application of Nav reduced the percentage of SA-β-Gal-positive cells after irradiation but not after pretreatment with Eto.In response to triggers of senescence,cultured Panc02 cells showed increased protein levels ofγH2AX and the autophagy marker LC3B-II,and higher mRNA levels of Cdkn1a,Mdm2,and PAI-1,while the effects of Nav were variable.Conclusions:In vitro and in vivo,the combination of senescence triggers with Nav inhibited tumor cell growth more effectively than the triggers alone.Our data also provide some evidence for senolytic effects of Nav in vitro.
