摘要
Objective Curcuma wenyujin,named Ezhu in Chinese,a traditional Chinese medicine,which has been shown to possess anticarcinogenic activity and used for the treatment of tumor in China,for example,hepatic cancer and leukemia.β-elemene,a component of Curcuma wenyujin,was largely reported as a main active component for anti-tumor effect of Curcuma wenyujin.However,furanodiene,another main component of Curcuma wenyujin,was seldom investigated for its anti-tumor activities.In the present study,we aim to investigate the effect of furanodiene on human leukemia HL60 cells and to study the accurate molecular mechanisms of its action.Methods Trypan blue exclusion experiment was used for cell growth inhibition assay.The apoptotic characterizations were assessed by flow cytometry analysis,AO/EB staining assay and agarose gel electrophoresis assay.The expression of apoptosis-related proteins and the release of cytochrome c from mitochondrial were detected by western blotting,and the mRNA levels of TNF-α and TNFR1 were probed by RT-PCR.Formation of TNFR1 complex was analyzed by using immunoprecipitation of TNFR1 with TRRF1/2 and RIP.Results Furanodiene(10-100 μM)treatment inhibited the growth of HL60 cells in a concentration-dependent manner.The effect of furanodiene on HL60 cells was associated with the induction of apoptosis,which was characterized by DNA fragmentation,cleavage of poly(ADP-ribose)polymerase(PARP),caspase-3,caspase-8 and caspase-9.In the Bcl-2 family proteins,Bid protein(a substrate of caspase-8)was activated by furanodiene,but Bcl-2,Bax and Bcl-xL proteins were not inuenced by furanodiene stimulation.Furanodiene elicited cytochrome c release from mitochondria into the cytosol.Moreover,furanodiene treatment caused the upregulation of TNFR1,the formation of TNFR1 complex and an obvious production of TNF-α in HL60 cells.The soluble TNFR1 receptor effectively inhibited furanodiene-induced apoptosis.Conclusions Furanodiene inhibited the growth of HL60 leukemia cells via induction of apoptosis.Furanodiene-induced apoptosis in HL60 cells is mediated by upregulation of TNF receptor 1 as well as induction of TNF-α production to activate TNFR1 signaling pathway.Our research provides insight into the molecular mechanisms on furanodiene-induced cell death,and may aid to the development of furanodiene as a new anti-tumor agent.
Objective Curcuma wenyujin,named Ezhu in Chinese,a traditional Chinese medicine,which has been shown to possess anticarcinogenic activity and used for the treatment of tumor in China,for example,hepatic cancer and leukemia.β-elemene,a component of Curcuma wenyujin,was largely reported as a main active component for anti-tumor effect of Curcuma wenyujin.However,furanodiene,another main component of Curcuma wenyujin,was seldom investigated for its anti-tumor activities.In the present study,we aim to investigate the effect of furanodiene on human leukemia HL60 cells and to study the accurate molecular mechanisms of its action.Methods Trypan blue exclusion experiment was used for cell growth inhibition assay.The apoptotic characterizations were assessed by flow cytometry analysis,AO/EB staining assay and agarose gel electrophoresis assay.The expression of apoptosis-related proteins and the release of cytochrome c from mitochondrial were detected by western blotting,and the mRNA levels of TNF-α and TNFR1 were probed by RT-PCR.Formation of TNFR1 complex was analyzed by using immunoprecipitation of TNFR1 with TRRF1/2 and RIP.Results Furanodiene(10-100 μM)treatment inhibited the growth of HL60 cells in a concentration-dependent manner.The effect of furanodiene on HL60 cells was associated with the induction of apoptosis,which was characterized by DNA fragmentation,cleavage of poly(ADP-ribose)polymerase(PARP),caspase-3,caspase-8 and caspase-9.In the Bcl-2 family proteins,Bid protein(a substrate of caspase-8)was activated by furanodiene,but Bcl-2,Bax and Bcl-xL proteins were not inuenced by furanodiene stimulation.Furanodiene elicited cytochrome c release from mitochondria into the cytosol.Moreover,furanodiene treatment caused the upregulation of TNFR1,the formation of TNFR1 complex and an obvious production of TNF-α in HL60 cells.The soluble TNFR1 receptor effectively inhibited furanodiene-induced apoptosis.Conclusions Furanodiene inhibited the growth of HL60 leukemia cells via induction of apoptosis.Furanodiene-induced apoptosis in HL60 cells is mediated by upregulation of TNF receptor 1 as well as induction of TNF-α production to activate TNFR1 signaling pathway.Our research provides insight into the molecular mechanisms on furanodiene-induced cell death,and may aid to the development of furanodiene as a new anti-tumor agent.
作者
MA En-long1,WANG Xiao-long1,LI Yan-chun1,TAI Wen-jiao1,LI Te1,GUO Tao2(1.Department of Pharmacology,Shenyang Pharmaceutical University,Shenyang 110016,China
2.Department of Pharmacy,General Hospital of Shenyang Military Region,Shenyang 110016,China)
出处
《沈阳药科大学学报》
CAS
CSCD
北大核心
2008年第S1期76-77,共2页
Journal of Shenyang Pharmaceutical University
