Resistance to cancer therapy continues to be a major limitation for the successful treatment of cancer. There are many published studies on therapy resistance in breast and prostate cancers; however, there are current...Resistance to cancer therapy continues to be a major limitation for the successful treatment of cancer. There are many published studies on therapy resistance in breast and prostate cancers; however, there are currently no data on molecular markers associated with resistance. The conflicting data were reported regarding the AKT/m-TOR signaling pathway components as markers predicting resistance. The AKT/m-TOR signaling pathway is involved in the development of many human cancers; its activation is related to cell proliferation, angiogenesis, apoptosis, as well as to therapy resistance. Molecular alterations in the AKT/m-TOR signaling pathway provide a platform to identify universal markers associated with the development of resistance to cancer therapy.展开更多
OBJECTIVE We previously showed that human pancreatic stellate cells(HPSCs)promote pancreatic ductal adenocarcinoma(PDAC)cell growth by activating nuclear factor erythroid2-related factor 2(Nrf2),a key transcriptional ...OBJECTIVE We previously showed that human pancreatic stellate cells(HPSCs)promote pancreatic ductal adenocarcinoma(PDAC)cell growth by activating nuclear factor erythroid2-related factor 2(Nrf2),a key transcriptional regulator of cytoprotective genes.We aim to investigate whether Nrf2-mediated metabolic reprogramming and reactive oxygen species(ROS)detoxification are involved in HPSCs-mediated cell growth.METHODS Nrf2-mediated metabolic genes expression of pentose phosphate pathway(PPP)for purine nucleotide synthesis;glutamine metabolism for nicotinamide adenine dinucleotide phosphate(NADPH)-equivalent producers and also glutathione biosynthesis both for intracellular ROS inactivation were examined using quantitative real-time PCR(qRT-PCR)after treated with conditioned media derived from HPSCs(HPSC-CM)in human PDAC cells(BxPC-3and AsPC-1)with or without Nrf2 gene silencing using siRNA-mediated technique.Metabolites involved in PPP for purine nucleotide and NADPH generation were selected and their concentration was measured using UHPLC-MS/MS.Antioxidants,tiron and N-acetylcysteine(NAC)were used to attenuate the intracellular ROS rendered by Nrf2 before measuring PDAC cell growth and also phosphorylation of extracellular signal-regulated kinase(ERK)1/2and protein kinase B(AKT)using MTT and Western blotting,respectively.RESULTS Metabolically,HPSC-CMupregulated Nrf2-mediated genes involved in three metabolic pathways(G6PD,PGD,TKT,PPAT,MTHFD2,ME1,IDH1,GCLC and GCLM)in BxPC-3and AsPC-1cells.HPSC-CM was able to upregulate all the metabolic genes after Nrf2 gene silencing,and also significantly increased the metabolite concentration of ribose 5-phosphate and inosine 5′-monophosphate,which are involved in nucleotide synthesis for cell growth.Decreasing the intracellular ROS rendered by Nrf2 suppressed PDAC cell growth and also phosphorylation of ERK 1/2and AKT protein.CONCLUSION Our findings reveal that HPSC-CM activates Nrf2-mediated metabolic reprogramming,which leads to purine nucleotide synthesis and ROS detoxification to promote PDAC cell growth.展开更多
Objective To observe the apoptosis of osteosarcoma MG-63 cells induced by As2O3 and to explore its possible mechanisms. Methods The flowcytometric analysis and transmission electronmicroscope were performed to investi...Objective To observe the apoptosis of osteosarcoma MG-63 cells induced by As2O3 and to explore its possible mechanisms. Methods The flowcytometric analysis and transmission electronmicroscope were performed to investigate the inducing apoptosis and inhibitative of As2O3 on osteosarcoma MG-63 cells. In order to study mechanism of apoptosis in MG-63 cells treated with As2 O3, microarray was performed. The down-regulated gene was confirmed by RT-PCR, Northern-blotting. Results After treated with As2O3, hypodiploid peak before G0/G1 phase was observed in MG-63 cells through FCM analysis. Loss of microvilli, condensation and fragmentation of nuclear chromatin, condensation of cytoplasmic organelles, dilatation of the endoplasmic reticulum shrinkage of cells and alterations in cell membranes and apoptosis bodies which were observed in MG-63 cells treated with As2O3 by transmission electronmicroscope. The results of microarray show that As2 O3 induced MG-63 cell apoptosis involves down-regulation of IEX-1 and the down-regulated gene is confirmed by RT-PCR and Northern-blotting.Conclusion The results show that As2 O3 selectively inhibits growth of the solid tumor MG-63 cells by triggering apoptosis and indicates MG-63 induced by As2O3 cell apoptosis may through the IEX-1 pathway.展开更多
北京协和医院公认是中国最好的医院,我们共同的想法是创立一个最好的转化医学中心,或许加利福尼亚大学旧金山分校（The University of California,SanFrancisco,UCSF）的作法不能机械照搬,但今天我将介绍一些在任何一片土壤上均能实现...北京协和医院公认是中国最好的医院,我们共同的想法是创立一个最好的转化医学中心,或许加利福尼亚大学旧金山分校（The University of California,SanFrancisco,UCSF）的作法不能机械照搬,但今天我将介绍一些在任何一片土壤上均能实现的基本架构,希望提供一些UCSF和北京协和医学院（Peking Union Medical College,PUMC）可以共同实现的目标。展开更多
Despite progresses achieved in the therapy of tumors, the prognosis of patients is still limited by reccurence of residual tumor cells. Cancer cell dormancy plays a pivotal role in cancer relapse and drug resistance. ...Despite progresses achieved in the therapy of tumors, the prognosis of patients is still limited by reccurence of residual tumor cells. Cancer cell dormancy plays a pivotal role in cancer relapse and drug resistance. In recent years, tumor cells undergoing EMT(epithelial-mesenchymal transition), CSCs(cancer stem cells) and CTCs(circulating tumor cells) are proved to share some common characteristics and show a cell cycle arrest phenotype. Thus, understanding the dormant stage of tumor cells could facilitate us in discovering ways to accelerate the development of tumor therapy and prevent its reccurence. In this review, we summarize the specific process of tumor cell dormancy induced by pharmacotherapy, and consider that dormancy is an initiative response rather than a passive defense to cytotoxicity. Besides, we probe into the mechanisms of tumor cell dormancy-mediated drug resistance, anticipating paving a way to target dormant tumor cells and result in better clinical outcomes.展开更多
HLCDG1, which locates in chromosome 5q33, is a novel gene cloned recently. The HLCDG1 expression was significantly down regulated in the primary lung carcinoma. It was previously studied that HLCDG1 acted like a tumor...HLCDG1, which locates in chromosome 5q33, is a novel gene cloned recently. The HLCDG1 expression was significantly down regulated in the primary lung carcinoma. It was previously studied that HLCDG1 acted like a tumor suppressor gene. In this paper, proteomics studies were performed to analyze the proteomic expression patterns in the HLCDG1-transfected human lung carcinoma cell line (A549-HLCDG1) and in the control vector-transfecred human lung carcinoma cell line (A549-vector). Employing two dimensional gel eleetrophoresis (2DE), the global pattern of protein expressions in A549-HLCDG1 human lung adenocarcinoma cell line expressing stably HL-CDG1 gene were compared with those of control A549-vector cell line to generate a differential protein expression catalog. Forty-two differentially expressed proteins were screened. Thirteen differential proteins were identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS), which were 6 upregulated (MSH5, MOD, MDH precursor, ETFβ, Prxd Ⅵ and JM23) and 7 downregulated (PLC-δ1, hnRNPA2,hnRNPB1, TIM, TCTP, nm23H-1 and PrxdⅤ) proteins in A549-HLCDG1 cells compared to control A549-vector cells. The above identified proteins were involved in energy metabolism, transcription regulation, antioxidation,cell cycle, metastasis, DNA methylation and mismatch repair. Therefore, these differential expression proteins by HLCDG1 transfection may play some important roles for investigation of the biochemical basis of growth suppression of HLCDG1 gene in lung carcinoma cells A549. Further understanding of this data base may provide valuable resources for the developing novel diagnostic markers and therapeutic targets of lung cancer.展开更多
文摘Resistance to cancer therapy continues to be a major limitation for the successful treatment of cancer. There are many published studies on therapy resistance in breast and prostate cancers; however, there are currently no data on molecular markers associated with resistance. The conflicting data were reported regarding the AKT/m-TOR signaling pathway components as markers predicting resistance. The AKT/m-TOR signaling pathway is involved in the development of many human cancers; its activation is related to cell proliferation, angiogenesis, apoptosis, as well as to therapy resistance. Molecular alterations in the AKT/m-TOR signaling pathway provide a platform to identify universal markers associated with the development of resistance to cancer therapy.
基金The project supported by University of Malaya HIR/MOHE/MED-12(Chung)
文摘OBJECTIVE We previously showed that human pancreatic stellate cells(HPSCs)promote pancreatic ductal adenocarcinoma(PDAC)cell growth by activating nuclear factor erythroid2-related factor 2(Nrf2),a key transcriptional regulator of cytoprotective genes.We aim to investigate whether Nrf2-mediated metabolic reprogramming and reactive oxygen species(ROS)detoxification are involved in HPSCs-mediated cell growth.METHODS Nrf2-mediated metabolic genes expression of pentose phosphate pathway(PPP)for purine nucleotide synthesis;glutamine metabolism for nicotinamide adenine dinucleotide phosphate(NADPH)-equivalent producers and also glutathione biosynthesis both for intracellular ROS inactivation were examined using quantitative real-time PCR(qRT-PCR)after treated with conditioned media derived from HPSCs(HPSC-CM)in human PDAC cells(BxPC-3and AsPC-1)with or without Nrf2 gene silencing using siRNA-mediated technique.Metabolites involved in PPP for purine nucleotide and NADPH generation were selected and their concentration was measured using UHPLC-MS/MS.Antioxidants,tiron and N-acetylcysteine(NAC)were used to attenuate the intracellular ROS rendered by Nrf2 before measuring PDAC cell growth and also phosphorylation of extracellular signal-regulated kinase(ERK)1/2and protein kinase B(AKT)using MTT and Western blotting,respectively.RESULTS Metabolically,HPSC-CMupregulated Nrf2-mediated genes involved in three metabolic pathways(G6PD,PGD,TKT,PPAT,MTHFD2,ME1,IDH1,GCLC and GCLM)in BxPC-3and AsPC-1cells.HPSC-CM was able to upregulate all the metabolic genes after Nrf2 gene silencing,and also significantly increased the metabolite concentration of ribose 5-phosphate and inosine 5′-monophosphate,which are involved in nucleotide synthesis for cell growth.Decreasing the intracellular ROS rendered by Nrf2 suppressed PDAC cell growth and also phosphorylation of ERK 1/2and AKT protein.CONCLUSION Our findings reveal that HPSC-CM activates Nrf2-mediated metabolic reprogramming,which leads to purine nucleotide synthesis and ROS detoxification to promote PDAC cell growth.
文摘Objective To observe the apoptosis of osteosarcoma MG-63 cells induced by As2O3 and to explore its possible mechanisms. Methods The flowcytometric analysis and transmission electronmicroscope were performed to investigate the inducing apoptosis and inhibitative of As2O3 on osteosarcoma MG-63 cells. In order to study mechanism of apoptosis in MG-63 cells treated with As2 O3, microarray was performed. The down-regulated gene was confirmed by RT-PCR, Northern-blotting. Results After treated with As2O3, hypodiploid peak before G0/G1 phase was observed in MG-63 cells through FCM analysis. Loss of microvilli, condensation and fragmentation of nuclear chromatin, condensation of cytoplasmic organelles, dilatation of the endoplasmic reticulum shrinkage of cells and alterations in cell membranes and apoptosis bodies which were observed in MG-63 cells treated with As2O3 by transmission electronmicroscope. The results of microarray show that As2 O3 induced MG-63 cell apoptosis involves down-regulation of IEX-1 and the down-regulated gene is confirmed by RT-PCR and Northern-blotting.Conclusion The results show that As2 O3 selectively inhibits growth of the solid tumor MG-63 cells by triggering apoptosis and indicates MG-63 induced by As2O3 cell apoptosis may through the IEX-1 pathway.
文摘北京协和医院公认是中国最好的医院,我们共同的想法是创立一个最好的转化医学中心,或许加利福尼亚大学旧金山分校（The University of California,SanFrancisco,UCSF）的作法不能机械照搬,但今天我将介绍一些在任何一片土壤上均能实现的基本架构,希望提供一些UCSF和北京协和医学院（Peking Union Medical College,PUMC）可以共同实现的目标。
基金supported by grants from The Fundamental ResearchFunds for the Central Universities(223000/862631)The National Natural Science Foundation of China(81773147)The Natural Science Foundation of Hunan Province,China(2015JJ2181)
文摘Despite progresses achieved in the therapy of tumors, the prognosis of patients is still limited by reccurence of residual tumor cells. Cancer cell dormancy plays a pivotal role in cancer relapse and drug resistance. In recent years, tumor cells undergoing EMT(epithelial-mesenchymal transition), CSCs(cancer stem cells) and CTCs(circulating tumor cells) are proved to share some common characteristics and show a cell cycle arrest phenotype. Thus, understanding the dormant stage of tumor cells could facilitate us in discovering ways to accelerate the development of tumor therapy and prevent its reccurence. In this review, we summarize the specific process of tumor cell dormancy induced by pharmacotherapy, and consider that dormancy is an initiative response rather than a passive defense to cytotoxicity. Besides, we probe into the mechanisms of tumor cell dormancy-mediated drug resistance, anticipating paving a way to target dormant tumor cells and result in better clinical outcomes.
基金Projects(30000074, 30471954) supported by the National Natural Science Foundation of China project(2003034467)supported by the Postdoctoral Science Foundation of China
文摘HLCDG1, which locates in chromosome 5q33, is a novel gene cloned recently. The HLCDG1 expression was significantly down regulated in the primary lung carcinoma. It was previously studied that HLCDG1 acted like a tumor suppressor gene. In this paper, proteomics studies were performed to analyze the proteomic expression patterns in the HLCDG1-transfected human lung carcinoma cell line (A549-HLCDG1) and in the control vector-transfecred human lung carcinoma cell line (A549-vector). Employing two dimensional gel eleetrophoresis (2DE), the global pattern of protein expressions in A549-HLCDG1 human lung adenocarcinoma cell line expressing stably HL-CDG1 gene were compared with those of control A549-vector cell line to generate a differential protein expression catalog. Forty-two differentially expressed proteins were screened. Thirteen differential proteins were identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS), which were 6 upregulated (MSH5, MOD, MDH precursor, ETFβ, Prxd Ⅵ and JM23) and 7 downregulated (PLC-δ1, hnRNPA2,hnRNPB1, TIM, TCTP, nm23H-1 and PrxdⅤ) proteins in A549-HLCDG1 cells compared to control A549-vector cells. The above identified proteins were involved in energy metabolism, transcription regulation, antioxidation,cell cycle, metastasis, DNA methylation and mismatch repair. Therefore, these differential expression proteins by HLCDG1 transfection may play some important roles for investigation of the biochemical basis of growth suppression of HLCDG1 gene in lung carcinoma cells A549. Further understanding of this data base may provide valuable resources for the developing novel diagnostic markers and therapeutic targets of lung cancer.
