Withaferin A (WA) is a bioactive compound derived from a medicinal plant Withania somnifera and has potential therapeutic effects against various types of cancers. The purpose of this study is to investigate an apopto...Withaferin A (WA) is a bioactive compound derived from a medicinal plant Withania somnifera and has potential therapeutic effects against various types of cancers. The purpose of this study is to investigate an apoptotic effect of WA and identify its molecular target in HSC-3 and HSC-4 human oral cancer cell lines using Trypan blue exclusion assay, DAPI staining and western blotting. WA inhibited cell viability and induced apoptosis in a concentration- or time-dependent manner, as evidenced by induction of nuclear condensation and fragmentation, activation of caspase 3 and poly (ADP-ribose) polymerase (PARP) cleavage. WA-induced apoptosis was partly diminished by Z-VAD, a pancaspase inhibitor. WA also increased Bim and Bax protein in HSC-3 and HSC-4 cells, respectively. These results suggest that WA may be a potential chemotherapeutic drug candidate against human oral cancer.展开更多
Glioblastoma(GBM) is the most common,malignant,and lethal primary brain tumor in adults.Up to now,there is no effective drug for GBM.Withaferin A(WFA) is mainly derived from Indian Winter cherry.It has been traditiona...Glioblastoma(GBM) is the most common,malignant,and lethal primary brain tumor in adults.Up to now,there is no effective drug for GBM.Withaferin A(WFA) is mainly derived from Indian Winter cherry.It has been traditionally used in ayurvedic medicine.WFA has wide range of pharmaco.logical activities including cardioprotective,anti-inflammatory,immuno-modulatory properties.Recently,WFA was reported to inhibit the growth of many cancer cells;however,the precise molecular mecha.nisms of its anti-cancer activities in GBM remain unclear.Here,we found that treatment of WFA in U251 and U87-MG glioma cells inhibited the cell proliferation,released the cellular LDH,decreased the DNA synthesis,and inhibited the migration,invasion,and colony formation of cells.WFA also in.creased the apoptotic rate of cells,decreased the mitochondrial membrane potential,arrested cell cy.cle at G_2/M,inhibited the activity of caspase 3/7,and increased the protein expression of cleaved-cas.pase 3,cleaved PARP in U251 and U87-MG cells.In addition,cell apoptosis induced by WFA was as.sociated with increasing level of Bim,Bad,P21,P53 and decreasing the level of p-CDK1,cyclin A and B.It was also shown that cell apoptosis induced by WFA was associated with P38 signal pathway.These results demonstrated that WFA induced mitochondrial dependent apoptosis in glioblastoma cells which was associated with arresting the cell cycle at G_2/M phase by P38 pathway.Taken together,our findings suggest that WFA might be a promising chemotherapy drug in the treatment of GBM.展开更多
Aim:As our understanding of cancer stem cell(CSC)biology improves,search for inhibitory agents of CSCs and metastatic CSCs(mCSCs)positive for CXCR4 is warranted.Withaferin A(WA),a withanolide extracted from the medici...Aim:As our understanding of cancer stem cell(CSC)biology improves,search for inhibitory agents of CSCs and metastatic CSCs(mCSCs)positive for CXCR4 is warranted.Withaferin A(WA),a withanolide extracted from the medicinal plant Withania somnifera,has been shown to exhibit anti-cancer effects through multiple mechanisms.Whether WA could selectively target CSCs,mCSCs,or non CSCs of a gastrointestinal(GI)carcinoma tumor remains unclear.Methods:Side-population(SP)analysis,flow cytometric phenotyping and sorting,non-invasive imaging in conjunction with xenotransplantation,and immunohistology were used in this investigation.Results:Using the lymph node metastatic GI cancer cell line UP-LN1,consisting of CD44^(high)/CD24^(low)floating(F)and CD44^(low)/CD24^(high)adherent(A)cell subsets,this study demonstrated that as compared with parental UP-LN1 cells or A cells,WA preferentially reduced F-cell proliferation,tumor sphere formation,and SP cells in vitro in greater effi ciencies by apoptosis.This action was mechanistically mediated via the down-regulation of CXCR4/CXCL12 and STAT3/interleukin-6 axes,both of which are instrumental in the acquisition of metastatic ability.Attenuation of interferon-γ-induced CXCR4 expression in F cells by knockdown with siRNA or blocking with an anti-CXCR4 antibody,followed by Western blot analysis,showed signifi cantly reduced metastatic potential in vitro.The extent of in vitro anti-invasive effect of WA on the IFN-γ-treated F cells was signifi cantly greater than on the F cells without WA treatment,or F cells treated with control siRNA or with control IgG antibody.The observed in vitro effects of WA on the CSC and mCSC targeting were validated by data obtained with non-invasive imaging in NOD/SCID mouse xenotransplantation.Conclusion:WA could effi ciently block the formation of both CSCs and mCSCs in the UP-LN1 cell line,suggesting that WA may be considered an effective therapeutic agent for this type of GI malignancies.展开更多
A combination therapy is discussed for the treatment of cancer, which has recently been applied successfully in a case study. The general approach is based on a combination of immune boosters, digestive enzymes and na...A combination therapy is discussed for the treatment of cancer, which has recently been applied successfully in a case study. The general approach is based on a combination of immune boosters, digestive enzymes and natural interferones. The idea is to stage a three-prong “pincer attack” on the tumor: while the immune boosters stimulate the immune system to attack cancer cells, the cytostatic properties of the interferones inhibit cancer growth, and the digestive enzymes accelerate the transport mechanism to remove the killed cancer cells from the body. The rationale of the therapy is explained, results of the case study are presented, and possible generalizations are mentioned.展开更多
文摘Withaferin A (WA) is a bioactive compound derived from a medicinal plant Withania somnifera and has potential therapeutic effects against various types of cancers. The purpose of this study is to investigate an apoptotic effect of WA and identify its molecular target in HSC-3 and HSC-4 human oral cancer cell lines using Trypan blue exclusion assay, DAPI staining and western blotting. WA inhibited cell viability and induced apoptosis in a concentration- or time-dependent manner, as evidenced by induction of nuclear condensation and fragmentation, activation of caspase 3 and poly (ADP-ribose) polymerase (PARP) cleavage. WA-induced apoptosis was partly diminished by Z-VAD, a pancaspase inhibitor. WA also increased Bim and Bax protein in HSC-3 and HSC-4 cells, respectively. These results suggest that WA may be a potential chemotherapeutic drug candidate against human oral cancer.
基金supported by National Natural Science Foundation of China(8157345481703536+2 种基金81703565) CAMS Innovation Fund for Medical Sciences(2016-I2M-3-007) Natural Science Foundation of Beijing(7172142)
文摘Glioblastoma(GBM) is the most common,malignant,and lethal primary brain tumor in adults.Up to now,there is no effective drug for GBM.Withaferin A(WFA) is mainly derived from Indian Winter cherry.It has been traditionally used in ayurvedic medicine.WFA has wide range of pharmaco.logical activities including cardioprotective,anti-inflammatory,immuno-modulatory properties.Recently,WFA was reported to inhibit the growth of many cancer cells;however,the precise molecular mecha.nisms of its anti-cancer activities in GBM remain unclear.Here,we found that treatment of WFA in U251 and U87-MG glioma cells inhibited the cell proliferation,released the cellular LDH,decreased the DNA synthesis,and inhibited the migration,invasion,and colony formation of cells.WFA also in.creased the apoptotic rate of cells,decreased the mitochondrial membrane potential,arrested cell cy.cle at G_2/M,inhibited the activity of caspase 3/7,and increased the protein expression of cleaved-cas.pase 3,cleaved PARP in U251 and U87-MG cells.In addition,cell apoptosis induced by WFA was as.sociated with increasing level of Bim,Bad,P21,P53 and decreasing the level of p-CDK1,cyclin A and B.It was also shown that cell apoptosis induced by WFA was associated with P38 signal pathway.These results demonstrated that WFA induced mitochondrial dependent apoptosis in glioblastoma cells which was associated with arresting the cell cycle at G_2/M phase by P38 pathway.Taken together,our findings suggest that WFA might be a promising chemotherapy drug in the treatment of GBM.
基金supported by grants from the National Science Council of Taiwan(NSC99-2314-B-038-032,NSC100-2314-B-038-016 to SK Liao)Chang Gung Medical Research Fund(CMRPD170043,CMRPG3A0811 to ST Pang and SK Liao).
文摘Aim:As our understanding of cancer stem cell(CSC)biology improves,search for inhibitory agents of CSCs and metastatic CSCs(mCSCs)positive for CXCR4 is warranted.Withaferin A(WA),a withanolide extracted from the medicinal plant Withania somnifera,has been shown to exhibit anti-cancer effects through multiple mechanisms.Whether WA could selectively target CSCs,mCSCs,or non CSCs of a gastrointestinal(GI)carcinoma tumor remains unclear.Methods:Side-population(SP)analysis,flow cytometric phenotyping and sorting,non-invasive imaging in conjunction with xenotransplantation,and immunohistology were used in this investigation.Results:Using the lymph node metastatic GI cancer cell line UP-LN1,consisting of CD44^(high)/CD24^(low)floating(F)and CD44^(low)/CD24^(high)adherent(A)cell subsets,this study demonstrated that as compared with parental UP-LN1 cells or A cells,WA preferentially reduced F-cell proliferation,tumor sphere formation,and SP cells in vitro in greater effi ciencies by apoptosis.This action was mechanistically mediated via the down-regulation of CXCR4/CXCL12 and STAT3/interleukin-6 axes,both of which are instrumental in the acquisition of metastatic ability.Attenuation of interferon-γ-induced CXCR4 expression in F cells by knockdown with siRNA or blocking with an anti-CXCR4 antibody,followed by Western blot analysis,showed signifi cantly reduced metastatic potential in vitro.The extent of in vitro anti-invasive effect of WA on the IFN-γ-treated F cells was signifi cantly greater than on the F cells without WA treatment,or F cells treated with control siRNA or with control IgG antibody.The observed in vitro effects of WA on the CSC and mCSC targeting were validated by data obtained with non-invasive imaging in NOD/SCID mouse xenotransplantation.Conclusion:WA could effi ciently block the formation of both CSCs and mCSCs in the UP-LN1 cell line,suggesting that WA may be considered an effective therapeutic agent for this type of GI malignancies.
文摘A combination therapy is discussed for the treatment of cancer, which has recently been applied successfully in a case study. The general approach is based on a combination of immune boosters, digestive enzymes and natural interferones. The idea is to stage a three-prong “pincer attack” on the tumor: while the immune boosters stimulate the immune system to attack cancer cells, the cytostatic properties of the interferones inhibit cancer growth, and the digestive enzymes accelerate the transport mechanism to remove the killed cancer cells from the body. The rationale of the therapy is explained, results of the case study are presented, and possible generalizations are mentioned.
