Objective:To assess if casticin induces caspase-mediated apoptosis via activation of mitochondrial pathway and upregulation of DR5 in human lung cancer ceils.Methods:Human non-small-cell lung carcinoma cell lines H460...Objective:To assess if casticin induces caspase-mediated apoptosis via activation of mitochondrial pathway and upregulation of DR5 in human lung cancer ceils.Methods:Human non-small-cell lung carcinoma cell lines H460,AS49 and H157 were cultured in vitro.The cytotoxic activities were determined using MTT assay.The apoptotic cells death was examined by flow cytometry using PI staining and DMA agarose gel electrophoresis.The activities of caspase-3, -8 and -9 were measured via ELISA.Cellular fractionation was determined by flow cytometry to assess release of cytochrome c and the mitochondrial transmembrane potential.Bcl-2/Bcl-XL/ XIAP/Bid/ DR5 and DR4 proteins were analyzed using western blot.Results:The concentrations required for a 50%decrease in cell growth(IC<sub>50</sub>) ranged from 1.8 to 3.2 Jt M.Casticin induced rapid apoptosis and triggered a series of effects associated with apoptosis by way of mitochondrial pathway,including the depolarization of the mitochondrial membrane,release of cytochrome c from mitochondria,activation of procaspase-9 and -3,and increase of DNA fragments.Moreover, the pan caspase inhibitor zVAD-FMK and the caspase-3 inhibitor zOEVD-FMK suppressed casticin-induced apoptosis.In addition,casticin induced XIAP and Bcl-XL down-regulation, Bax upregulation and Bid clearage.In H157 cell line,casticin increased expression of DRS at protein levels but not affect the expression of DR4.The prelreatmenl with DR5/Fc chimera protein effectively attenuated casticin-induced apoptosis in H157 cells.No correlation was found between cell sensitivity to casticin and that to p53 status,suggesting that casticin induce a p53- independent apoptosis.Conclusions:Our results demonstrate that casticin induces caspase-mediated apoptosis via activation of mitochondrial pathway and upregulation of DRS in human lung cancer cells.展开更多
Pancreatic cancer has become the fourth leading cause of cancer death in the last two decades. Only 3%-15% of patients diagnosed with pancreatic cancer had 5 year survival rate. Drug resistance, high metastasis, poor ...Pancreatic cancer has become the fourth leading cause of cancer death in the last two decades. Only 3%-15% of patients diagnosed with pancreatic cancer had 5 year survival rate. Drug resistance, high metastasis, poor prognosis and tumour relapse contributed to the malignancies and difficulties in treating pancreatic cancer. The current standard chemotherapy for pancreatic cancer is gemcitabine, however its efficacy is far from satisfactory, one of the reasons is due to the complex tumour microenvironment which decreases effective drug delivery to target cancer cell. Studies of the molecular pathology of pancreatic cancer have revealed that activation of KRAS, overexpression of cyclooxygenase-2, inactivation of p16<sup>INK4A</sup> and loss of p53 activities occurred in pancreatic cancer. Co-administration of gemcitabine and targeting the molecular pathological events happened in pancreatic cancer has brought an enhanced therapeutic effectiveness of gemcitabine. Therefore, studies looking for novel targets in hindering pancreatic tumour growth are emerging rapidly. In order to give a better understanding of the current findings and to seek the direction in future pancreatic cancer research; in this review we will focus on targets suppressing tumour metastatsis and progression, KRAS activated downstream effectors, the relationship of Notch signaling and Nodal/Activin signaling with pancreatic cancer cells, the current findings of non-coding RNAs in inhibiting pancreatic cancer cell proliferation, brief discussion in transcription remodeling by epigenetic modifiers (e.g., HDAC, BMI1, EZH2) and the plausible therapeutic applications of cancer stem cell and hyaluronan in tumour environment.展开更多
基金supported,in part,by grants from the National Natural Scientific Foundation of China(Nos.30760248, 81072161,81172139,81060183)Programs for Changjiang Scholars and Innovative Research Team in University(No. IRT1119)+4 种基金Programs for Guangxi Innovative Research Team (No.2011GXNSFF018005)Program of Science and Technology of Guangxi(No.1140003A-16)Project of Scientific Research of Hunan Province the Administration Bureau of Traditional Chinese Medicine(No.2010081)the Project of Scientific Research of Hunan Province the Department of Education (No.10C0975Major Project of Scientific Research of Hunan Province the Department of Education(No.09A054)
文摘Objective:To assess if casticin induces caspase-mediated apoptosis via activation of mitochondrial pathway and upregulation of DR5 in human lung cancer ceils.Methods:Human non-small-cell lung carcinoma cell lines H460,AS49 and H157 were cultured in vitro.The cytotoxic activities were determined using MTT assay.The apoptotic cells death was examined by flow cytometry using PI staining and DMA agarose gel electrophoresis.The activities of caspase-3, -8 and -9 were measured via ELISA.Cellular fractionation was determined by flow cytometry to assess release of cytochrome c and the mitochondrial transmembrane potential.Bcl-2/Bcl-XL/ XIAP/Bid/ DR5 and DR4 proteins were analyzed using western blot.Results:The concentrations required for a 50%decrease in cell growth(IC<sub>50</sub>) ranged from 1.8 to 3.2 Jt M.Casticin induced rapid apoptosis and triggered a series of effects associated with apoptosis by way of mitochondrial pathway,including the depolarization of the mitochondrial membrane,release of cytochrome c from mitochondria,activation of procaspase-9 and -3,and increase of DNA fragments.Moreover, the pan caspase inhibitor zVAD-FMK and the caspase-3 inhibitor zOEVD-FMK suppressed casticin-induced apoptosis.In addition,casticin induced XIAP and Bcl-XL down-regulation, Bax upregulation and Bid clearage.In H157 cell line,casticin increased expression of DRS at protein levels but not affect the expression of DR4.The prelreatmenl with DR5/Fc chimera protein effectively attenuated casticin-induced apoptosis in H157 cells.No correlation was found between cell sensitivity to casticin and that to p53 status,suggesting that casticin induce a p53- independent apoptosis.Conclusions:Our results demonstrate that casticin induces caspase-mediated apoptosis via activation of mitochondrial pathway and upregulation of DRS in human lung cancer cells.
文摘Pancreatic cancer has become the fourth leading cause of cancer death in the last two decades. Only 3%-15% of patients diagnosed with pancreatic cancer had 5 year survival rate. Drug resistance, high metastasis, poor prognosis and tumour relapse contributed to the malignancies and difficulties in treating pancreatic cancer. The current standard chemotherapy for pancreatic cancer is gemcitabine, however its efficacy is far from satisfactory, one of the reasons is due to the complex tumour microenvironment which decreases effective drug delivery to target cancer cell. Studies of the molecular pathology of pancreatic cancer have revealed that activation of KRAS, overexpression of cyclooxygenase-2, inactivation of p16<sup>INK4A</sup> and loss of p53 activities occurred in pancreatic cancer. Co-administration of gemcitabine and targeting the molecular pathological events happened in pancreatic cancer has brought an enhanced therapeutic effectiveness of gemcitabine. Therefore, studies looking for novel targets in hindering pancreatic tumour growth are emerging rapidly. In order to give a better understanding of the current findings and to seek the direction in future pancreatic cancer research; in this review we will focus on targets suppressing tumour metastatsis and progression, KRAS activated downstream effectors, the relationship of Notch signaling and Nodal/Activin signaling with pancreatic cancer cells, the current findings of non-coding RNAs in inhibiting pancreatic cancer cell proliferation, brief discussion in transcription remodeling by epigenetic modifiers (e.g., HDAC, BMI1, EZH2) and the plausible therapeutic applications of cancer stem cell and hyaluronan in tumour environment.
