Objective Tumor necrosis factor-related apoptosis-inducing ligand （TRAIL） resistance greatly limits the clinical therapeutic efficacy of TRAIL. Elucidating the molecular mechanism underlying TRAIL resistance will be...Objective Tumor necrosis factor-related apoptosis-inducing ligand （TRAIL） resistance greatly limits the clinical therapeutic efficacy of TRAIL. Elucidating the molecular mechanism underlying TRAIL resistance will be fundamental to resolving this problem. Methods Nuclear and cytoplasmic protein extraction and immunofluorescence （IF） assay were used to detect changes in heterogeneous nuclear ribonucleoprotein K （hnRNPK） localization in H1299 cells. The evaluation of cell apoptosis in cells transfected with GFP-hnRNPK, GFP-hnRNPK S284/353A, or GFP-hnRNPK S284/353D mutant was performed using cleaved caspase-3 antibody. The gene expression of XIAP was tested by quantitative RT-PCR. Results Previously, we reported that hnRNPK antagonized TRAIL-induced apoptosis through inhibition of PKC-mediated GSK313 phosphorylation. In this study, we further demonstrate that TRAIL treatment induces cytoplasmic accumulation of hnRNPK in H1299 cells. The hnRNPK localized in the cytoplasm has a higher capacity to antagonize TRAIL-induced apoptosis. Both ERK1/2 signaling inhibitor U0126 and ERK-phosphoacceptor-site mutant （GFP-hnRNPK S284/353A） diminish cytoplasmic accumulation of hnRNPK induced by TRAIL. Moreover, we show that XlAP is involved in hnRNPK-mediated TRAIL resistance in H2299 cells. Conclusion Taken together, these results give new insights into the understanding of the molecular mechanism associated with TRAIL resistance in lung adenocarcinoma.展开更多
Upon activation, naive T-helper cells can differentiate into two major distinct subsets, T helper 1 (Th1) and T helper 2 (Th2), as defined by their effector functions and cytokine secretion patterns. Cytokine milieu a...Upon activation, naive T-helper cells can differentiate into two major distinct subsets, T helper 1 (Th1) and T helper 2 (Th2), as defined by their effector functions and cytokine secretion patterns. Cytokine milieu and costimulatory molecules have been shown to play an essential role in determining T helper differentiation. However, it is still unclear how the effects of signals of co-stimulatory molecules and cytokines are exerted during T helper differentiation. We show evidence suggesting that while cytokine signals initiate differentiation program, the selective action of death effectors determines the endpoint balance of differenti-展开更多
目的:探讨肿瘤坏死因子相关凋亡诱导配体(TNF related apoptosis indncing ligmard,TRAIL)联合化疗药物顺铂(Cisplatin,DDP)、5氟尿嘧啶(5-Fluorouracil,5-FU)作用于鼻咽癌HNE-1细胞株后对其细胞毒作用及凋亡影响的研究。方法:MTT法检...目的:探讨肿瘤坏死因子相关凋亡诱导配体(TNF related apoptosis indncing ligmard,TRAIL)联合化疗药物顺铂(Cisplatin,DDP)、5氟尿嘧啶(5-Fluorouracil,5-FU)作用于鼻咽癌HNE-1细胞株后对其细胞毒作用及凋亡影响的研究。方法:MTT法检测其增殖抑制率,FCM法检测细胞凋亡率。结果:TRAIL与DDP、5-FU联合作用于HNE-1细胞株时,其抑制率、凋亡率高于单独作用时的抑制率和凋亡率。结论:TRAIL与5-FU、DDP联合作用有协同抑制HNE-1细胞株增殖、诱导HNE-1细胞株凋亡的作用。展开更多
基金supported by National Program on Key Basic Research Project(973 Program)(Grant No.2011CB910700)Natural Science Foundation of Guangdong Province(2016A030313083,2016A030313420)+2 种基金Guangzhou Science and Technology Project(20160701175)Fundamental Research Funds for the Central Universities(Grant No.21615407,21609317)Science and Technology Program of Huadu District of Guangzhou,Guangdong Province(15-HDWS-016)
文摘Objective Tumor necrosis factor-related apoptosis-inducing ligand （TRAIL） resistance greatly limits the clinical therapeutic efficacy of TRAIL. Elucidating the molecular mechanism underlying TRAIL resistance will be fundamental to resolving this problem. Methods Nuclear and cytoplasmic protein extraction and immunofluorescence （IF） assay were used to detect changes in heterogeneous nuclear ribonucleoprotein K （hnRNPK） localization in H1299 cells. The evaluation of cell apoptosis in cells transfected with GFP-hnRNPK, GFP-hnRNPK S284/353A, or GFP-hnRNPK S284/353D mutant was performed using cleaved caspase-3 antibody. The gene expression of XIAP was tested by quantitative RT-PCR. Results Previously, we reported that hnRNPK antagonized TRAIL-induced apoptosis through inhibition of PKC-mediated GSK313 phosphorylation. In this study, we further demonstrate that TRAIL treatment induces cytoplasmic accumulation of hnRNPK in H1299 cells. The hnRNPK localized in the cytoplasm has a higher capacity to antagonize TRAIL-induced apoptosis. Both ERK1/2 signaling inhibitor U0126 and ERK-phosphoacceptor-site mutant （GFP-hnRNPK S284/353A） diminish cytoplasmic accumulation of hnRNPK induced by TRAIL. Moreover, we show that XlAP is involved in hnRNPK-mediated TRAIL resistance in H2299 cells. Conclusion Taken together, these results give new insights into the understanding of the molecular mechanism associated with TRAIL resistance in lung adenocarcinoma.
文摘Upon activation, naive T-helper cells can differentiate into two major distinct subsets, T helper 1 (Th1) and T helper 2 (Th2), as defined by their effector functions and cytokine secretion patterns. Cytokine milieu and costimulatory molecules have been shown to play an essential role in determining T helper differentiation. However, it is still unclear how the effects of signals of co-stimulatory molecules and cytokines are exerted during T helper differentiation. We show evidence suggesting that while cytokine signals initiate differentiation program, the selective action of death effectors determines the endpoint balance of differenti-
文摘目的:探讨肿瘤坏死因子相关凋亡诱导配体(TNF related apoptosis indncing ligmard,TRAIL)联合化疗药物顺铂(Cisplatin,DDP)、5氟尿嘧啶(5-Fluorouracil,5-FU)作用于鼻咽癌HNE-1细胞株后对其细胞毒作用及凋亡影响的研究。方法:MTT法检测其增殖抑制率,FCM法检测细胞凋亡率。结果:TRAIL与DDP、5-FU联合作用于HNE-1细胞株时,其抑制率、凋亡率高于单独作用时的抑制率和凋亡率。结论:TRAIL与5-FU、DDP联合作用有协同抑制HNE-1细胞株增殖、诱导HNE-1细胞株凋亡的作用。
