Objectives:Chemotherapy-induced lung inflammation limits the efficacy of anticancer therapies such as gefitinib in non-small cell lung cancer(NSCLC).Glutamic acid-leucine-arginine positive(ELR+)CXC chemokines and thei...Objectives:Chemotherapy-induced lung inflammation limits the efficacy of anticancer therapies such as gefitinib in non-small cell lung cancer(NSCLC).Glutamic acid-leucine-arginine positive(ELR+)CXC chemokines and their receptors,CXC chemokine receptor 1 and 2(CXCR1 and CXCR2),mediate both inflammatory responses and tumor progression.This study evaluated the effects of CXCR1/2 antagonism by G31P,a CXC motif chemokine ligand 8(CXCL8)-mutated peptide,alone or in combination with gefitinib,on lung cancer growth and chemotherapyinduced pulmonary inflammation.Methods:Human NSCLC cell lines(A549 and H460)were treated with gefitinib and/or G31P.Cell proliferation,apoptosis,and signaling pathways,including protein kinase B(AKT)and extracellular signal-regulated kinase(ERK)phosphorylation,were evaluated by cell counting kit-8(CCK-8)assay,flow cytometry,and Western blotting.An orthotopic lung tumor xenograft model was established in BALB/c nude mice to evaluate tumor growth,metastasis,cytokine expression,and lung histopathology.A bleomycin-induced lung injury model was also used to assess the anti-inflammatory effects of G31P,with or without gefitinib,by quantitative reverse transcription polymerase chain reaction(qRT-PCR)and flowcytometry of inflammatorymarkers.Results:G31P andGefitinib,either alone or combined,inhibited proliferation and migration of A549 and H460 cells in vitro.Combination treatment effectively reducedAKT and ERKphosphorylation in both cell lines.In vivo,G31Pwith gefitinib significantly suppressed tumor growth,metastasis,and increased apoptosis.G31P decreased CXCL1 and CXCL2,and tumor necrosis factoralpha(TNF-α)mRNA levels,lung hydroxyproline content,and myeloperoxidase(MPO)activity in the lungs of mice.In the bleomycin-induced lung injury model,G31P similarly reduced inflammatory responses.Conclusion:CXCR1/2 antagonismby G31P attenuates chemotherapy-induced pulmonary inflammation and enhances the anti-tumor efficacy of gefitinib in NSCLC.These findings support the therapeutic potential of G31P as an adjuvant to epidermal growth factor receptor tyrosine kinase inhibitors(EGFR-TKIs)to improve clinical outcomes by limiting inflammation.展开更多
Background:KIT proto-oncogene,receptor tyrosine kinase(KIT,CD117)and platelet-derived growth factor-alpha(PDGFRA)are key drivers of gastrointestinal stromal tumors(GIST),but resistance to targeted therapy often arises...Background:KIT proto-oncogene,receptor tyrosine kinase(KIT,CD117)and platelet-derived growth factor-alpha(PDGFRA)are key drivers of gastrointestinal stromal tumors(GIST),but resistance to targeted therapy often arises from tumor protein p53(p53)alterations and loss of cell cycle control.However,the role of p53 status in GIST therapeutic potential has rarely been studied,so this study aimed to employ both wild-type andmutant p53 GIST models to investigate how p53 dysfunction influences the efficacy of p53 pathway-targeted therapies.Methods:The efficacy of the mouse double minute 2 homolog(MDM2)inhibitor(HDM201)and the Wee1 G2 checkpoint kinase(Wee1)inhibitor(adavosertib)was confirmed in both p53 wild-type(p53 WT)and p53 mutant(p53 MT)GIST cells.The anti-proliferative effects were assessed using the Cell Counting Kit-8(CCK-8)assay.Flow cytometry(FACS)and immunoblotting were employed to evaluate apoptosis and the expression of proteins related to drug efficacy.These findings were further validated in a xenograft model.Results:HDM201 selectively inhibited growth and triggered apoptosis in p53WT GIST cells,while adavosertib was effective mainly in p53 MT cells.Western blot analysis revealed thatHDM201 increased p53 and p21 levels in p53WT cells,and adavosertib affectedWee1 and phospho-cdc2 expression in both p53WT and p53 MT cells.In a xenograft mouse model,HDM201 significantly reduced the tumor volume and weight in p53WTGIST cells,whereas p53MT tumors showed only a moderate size reduction with adavosertib,without significant changes.Conclusions:Our results highlight the importance of p53 status in guiding GIST treatment.p53 WT tumors respond toMDM2 inhibitors,while p53 MTtumors show greater sensitivity toWee1 inhibitors,supporting p53 pathway targeting as a promising strategy for GIST patients.展开更多
Although previous studies showed that the principal oncoprotein encoded by Epstein-Barr virus, latent membrane protein 1 (LMP1), could induce the nasopharyngeal carcinoma cells in G2/M phase increased, littleis known ...Although previous studies showed that the principal oncoprotein encoded by Epstein-Barr virus, latent membrane protein 1 (LMP1), could induce the nasopharyngeal carcinoma cells in G2/M phase increased, littleis known about the target molecules and mechanisms. The present study demonstrated that LMP1 couldinduce the accumulation of p53 protein and upregulate its transactivity in a dose dependent manner, whichresulted in the decrease of the kinase activity of cdc2/cyclin B complex and inducing arrest at G2/M phasethrough the activation of NF-kB and AP-1 signaling pathways, and the effect of NF-kB was more obviousthan that of AP-1. This study provided some significant evidence for further elucidating the molecular mechanisms that LMP1 had effects on the surveillance mechanism of cell cycle and promoting the survivalof transformed cells and tumorigenesis.展开更多
Previous work has demonstrated that the sensitization of spinal neurons and microglia is important in the development of pain behaviors induced by BmK I,a Na^+ channel activator and a major peptide component of the ve...Previous work has demonstrated that the sensitization of spinal neurons and microglia is important in the development of pain behaviors induced by BmK I,a Na^+ channel activator and a major peptide component of the venom of the scorpion Buthus martensi Karsch(BmK).We found that the expression of P2X7 receptors(P2X7Rs)was up-regulated in the ipsilateral spinal dorsal horn after BmK I injection in rats.P2X7R was selectively localized in microglia but not astrocytes or neurons.Similarly,interleukin 1β(IL-1β)was selectively up-regulated in microglia in the spinal dorsal horn after BmK I injection.Intrathecal injection of P2X7R antagonists largely reduced BmK I-induced spontaneous and evoked pain behaviors,and the up-regulation of P2X7R and IL-1β in the spinal cord.These data suggested that the up-regulation of P2X7Rs mediates microglial activation in the spinal dorsal horn,and therefore contributes to the development of BmK I-induced pain.展开更多
目的:验证G2/S期应答相关蛋白1(G2 and S phase-expressed protein 1,GTSE1)在原发性肝癌组织及细胞株中高表达,并探讨其表达下调对肝癌细胞凋亡的影响。方法:采用内置R程序对GEO(Gene Expression Omnibus)数据库中4项原发性肝癌临床研...目的:验证G2/S期应答相关蛋白1(G2 and S phase-expressed protein 1,GTSE1)在原发性肝癌组织及细胞株中高表达,并探讨其表达下调对肝癌细胞凋亡的影响。方法:采用内置R程序对GEO(Gene Expression Omnibus)数据库中4项原发性肝癌临床研究的基因表达矩阵数据进行分析,筛选肝癌与癌旁组织差异表达基因。采用蛋白质印迹法验证3例肝癌组织和3株肝癌细胞中GTSE1蛋白表达水平。向肝癌Hep-G2和Bel-7402细胞中分别转染GTSE1 siRNA后,采用CCK-8和FCM法分别检测GTSE1表达下调对肝癌细胞增殖、凋亡及5-氟尿嘧啶(5-fluorouracil,5-FU)敏感性的影响,进一步采用蛋白质印迹法检测p53含量及其亚细胞定位,以及Bcl-2家族成员的表达变化。结果:4项研究中原发性肝癌组织表达水平均上调和下调的基因分别为51和146个。其中,上调基因产物GTSE1蛋白在3例临床肝癌组织标本和3株肝癌细胞Hep-G2、Bel-7402和SMMC-7721中均得到进一步验证(P值均<0.001)。siRNA转染后,肝癌Hep-G2和Bel-7402细胞中GTSE1基因沉默效率>70%。与对照组相比,GTSE1表达下调后肝癌Hep-G2和Bel-7402细胞增殖被明显抑制,并随着时间延长,抑制作用逐渐增强(P值均<0.05);GTSE1表达下调后肝癌Hep-G2和Bel-7402细胞周期被阻滞在G1期,同时细胞凋亡率明显升高,且细胞对5-FU的敏感性明显增强(P值均<0.01)。GTSE1表达下调后的肝癌Hep-G2和Bel-7402细胞中总p53及磷酸化P53表达水平均明显上调(P值均<0.05),且细胞核内p53水平升高(P值均<0.01);Bcl-2家族促凋亡成员Bax和Bak表达水平均上调,而抗凋亡成员Bcl-2表达水平下调(P值均<0.05)。结论:下调肝癌中GTSE1蛋白表达能通过活化p53通路促进细胞凋亡,并增加肝癌细胞对化疗药物的敏感性。展开更多
基金supported by grants from the National Natural Science Foundation of China,No.U1604174Health Commission of Henan Province-Province and Ministry Co-Construction Project,Nos.20170123,SBGJ202002004Health Commission of Henan Province-Research Innovative Talents Project(51282).
文摘Objectives:Chemotherapy-induced lung inflammation limits the efficacy of anticancer therapies such as gefitinib in non-small cell lung cancer(NSCLC).Glutamic acid-leucine-arginine positive(ELR+)CXC chemokines and their receptors,CXC chemokine receptor 1 and 2(CXCR1 and CXCR2),mediate both inflammatory responses and tumor progression.This study evaluated the effects of CXCR1/2 antagonism by G31P,a CXC motif chemokine ligand 8(CXCL8)-mutated peptide,alone or in combination with gefitinib,on lung cancer growth and chemotherapyinduced pulmonary inflammation.Methods:Human NSCLC cell lines(A549 and H460)were treated with gefitinib and/or G31P.Cell proliferation,apoptosis,and signaling pathways,including protein kinase B(AKT)and extracellular signal-regulated kinase(ERK)phosphorylation,were evaluated by cell counting kit-8(CCK-8)assay,flow cytometry,and Western blotting.An orthotopic lung tumor xenograft model was established in BALB/c nude mice to evaluate tumor growth,metastasis,cytokine expression,and lung histopathology.A bleomycin-induced lung injury model was also used to assess the anti-inflammatory effects of G31P,with or without gefitinib,by quantitative reverse transcription polymerase chain reaction(qRT-PCR)and flowcytometry of inflammatorymarkers.Results:G31P andGefitinib,either alone or combined,inhibited proliferation and migration of A549 and H460 cells in vitro.Combination treatment effectively reducedAKT and ERKphosphorylation in both cell lines.In vivo,G31Pwith gefitinib significantly suppressed tumor growth,metastasis,and increased apoptosis.G31P decreased CXCL1 and CXCL2,and tumor necrosis factoralpha(TNF-α)mRNA levels,lung hydroxyproline content,and myeloperoxidase(MPO)activity in the lungs of mice.In the bleomycin-induced lung injury model,G31P similarly reduced inflammatory responses.Conclusion:CXCR1/2 antagonismby G31P attenuates chemotherapy-induced pulmonary inflammation and enhances the anti-tumor efficacy of gefitinib in NSCLC.These findings support the therapeutic potential of G31P as an adjuvant to epidermal growth factor receptor tyrosine kinase inhibitors(EGFR-TKIs)to improve clinical outcomes by limiting inflammation.
基金financially supported by grants from the Chang-Gung Memorial Hospital(CMRPG3J0971~3,CMRPVVP0111,and CMRPVVQ0041 to CEWCMRPG3P0101 to HJS)the National Science and Technology Council(113-2628-B-182-001-MY3 and 113-2811-B-182-024 to CEW).
文摘Background:KIT proto-oncogene,receptor tyrosine kinase(KIT,CD117)and platelet-derived growth factor-alpha(PDGFRA)are key drivers of gastrointestinal stromal tumors(GIST),but resistance to targeted therapy often arises from tumor protein p53(p53)alterations and loss of cell cycle control.However,the role of p53 status in GIST therapeutic potential has rarely been studied,so this study aimed to employ both wild-type andmutant p53 GIST models to investigate how p53 dysfunction influences the efficacy of p53 pathway-targeted therapies.Methods:The efficacy of the mouse double minute 2 homolog(MDM2)inhibitor(HDM201)and the Wee1 G2 checkpoint kinase(Wee1)inhibitor(adavosertib)was confirmed in both p53 wild-type(p53 WT)and p53 mutant(p53 MT)GIST cells.The anti-proliferative effects were assessed using the Cell Counting Kit-8(CCK-8)assay.Flow cytometry(FACS)and immunoblotting were employed to evaluate apoptosis and the expression of proteins related to drug efficacy.These findings were further validated in a xenograft model.Results:HDM201 selectively inhibited growth and triggered apoptosis in p53WT GIST cells,while adavosertib was effective mainly in p53 MT cells.Western blot analysis revealed thatHDM201 increased p53 and p21 levels in p53WT cells,and adavosertib affectedWee1 and phospho-cdc2 expression in both p53WT and p53 MT cells.In a xenograft mouse model,HDM201 significantly reduced the tumor volume and weight in p53WTGIST cells,whereas p53MT tumors showed only a moderate size reduction with adavosertib,without significant changes.Conclusions:Our results highlight the importance of p53 status in guiding GIST treatment.p53 WT tumors respond toMDM2 inhibitors,while p53 MTtumors show greater sensitivity toWee1 inhibitors,supporting p53 pathway targeting as a promising strategy for GIST patients.
文摘目的:探讨CXCR1/CXCR2受体拮抗剂-G31P对人前列腺癌细胞PC-3增殖的体内外抑制作用。方法:采用CCK-8法研究不同浓度G31P对PC-3细胞体外增殖的抑制作用。建立体内绿色荧光蛋白(Green fluorescent protein,GFP)标记人雄激素非依赖性前列腺癌细胞PC-3裸鼠原位移植瘤模型,观察G31P对裸鼠前列腺癌原位移植瘤的体积、重量的影响。结果:CCK-8结果显示100 ng/ml G31P与对照组相比分别作用1、3 d和5 d差异具有统计学意义(1 d P=0.007、3 d P=0.001、5 d P=0.028,均为P<0.05)。前列腺癌PC-3细胞裸鼠模型体内实验显示,与对照组(100μl N.S)相比,G31P处理组(0.5 mg/kg)从给药第18天起能显著抑制前列腺肿瘤的体积(P=0.026,P<0.05);与对照组相比G31P处理组在抑制前列腺肿瘤重量方面有明显作用(P=0.027,P<0.05)。结论:G31P体内外实验均能抑制人雄激素非依赖性前列腺癌细胞系PC-3的增殖。
文摘Although previous studies showed that the principal oncoprotein encoded by Epstein-Barr virus, latent membrane protein 1 (LMP1), could induce the nasopharyngeal carcinoma cells in G2/M phase increased, littleis known about the target molecules and mechanisms. The present study demonstrated that LMP1 couldinduce the accumulation of p53 protein and upregulate its transactivity in a dose dependent manner, whichresulted in the decrease of the kinase activity of cdc2/cyclin B complex and inducing arrest at G2/M phasethrough the activation of NF-kB and AP-1 signaling pathways, and the effect of NF-kB was more obviousthan that of AP-1. This study provided some significant evidence for further elucidating the molecular mechanisms that LMP1 had effects on the surveillance mechanism of cell cycle and promoting the survivalof transformed cells and tumorigenesis.
基金supported by grants from the National Natural Science Foundation of China (31571032 and 31771191)supported by an Indiana Spinal Cord and Brain Injury Research Fund grant from Indiana State Department of Health, USA (2017)
文摘Previous work has demonstrated that the sensitization of spinal neurons and microglia is important in the development of pain behaviors induced by BmK I,a Na^+ channel activator and a major peptide component of the venom of the scorpion Buthus martensi Karsch(BmK).We found that the expression of P2X7 receptors(P2X7Rs)was up-regulated in the ipsilateral spinal dorsal horn after BmK I injection in rats.P2X7R was selectively localized in microglia but not astrocytes or neurons.Similarly,interleukin 1β(IL-1β)was selectively up-regulated in microglia in the spinal dorsal horn after BmK I injection.Intrathecal injection of P2X7R antagonists largely reduced BmK I-induced spontaneous and evoked pain behaviors,and the up-regulation of P2X7R and IL-1β in the spinal cord.These data suggested that the up-regulation of P2X7Rs mediates microglial activation in the spinal dorsal horn,and therefore contributes to the development of BmK I-induced pain.
文摘目的:验证G2/S期应答相关蛋白1(G2 and S phase-expressed protein 1,GTSE1)在原发性肝癌组织及细胞株中高表达,并探讨其表达下调对肝癌细胞凋亡的影响。方法:采用内置R程序对GEO(Gene Expression Omnibus)数据库中4项原发性肝癌临床研究的基因表达矩阵数据进行分析,筛选肝癌与癌旁组织差异表达基因。采用蛋白质印迹法验证3例肝癌组织和3株肝癌细胞中GTSE1蛋白表达水平。向肝癌Hep-G2和Bel-7402细胞中分别转染GTSE1 siRNA后,采用CCK-8和FCM法分别检测GTSE1表达下调对肝癌细胞增殖、凋亡及5-氟尿嘧啶(5-fluorouracil,5-FU)敏感性的影响,进一步采用蛋白质印迹法检测p53含量及其亚细胞定位,以及Bcl-2家族成员的表达变化。结果:4项研究中原发性肝癌组织表达水平均上调和下调的基因分别为51和146个。其中,上调基因产物GTSE1蛋白在3例临床肝癌组织标本和3株肝癌细胞Hep-G2、Bel-7402和SMMC-7721中均得到进一步验证(P值均<0.001)。siRNA转染后,肝癌Hep-G2和Bel-7402细胞中GTSE1基因沉默效率>70%。与对照组相比,GTSE1表达下调后肝癌Hep-G2和Bel-7402细胞增殖被明显抑制,并随着时间延长,抑制作用逐渐增强(P值均<0.05);GTSE1表达下调后肝癌Hep-G2和Bel-7402细胞周期被阻滞在G1期,同时细胞凋亡率明显升高,且细胞对5-FU的敏感性明显增强(P值均<0.01)。GTSE1表达下调后的肝癌Hep-G2和Bel-7402细胞中总p53及磷酸化P53表达水平均明显上调(P值均<0.05),且细胞核内p53水平升高(P值均<0.01);Bcl-2家族促凋亡成员Bax和Bak表达水平均上调,而抗凋亡成员Bcl-2表达水平下调(P值均<0.05)。结论:下调肝癌中GTSE1蛋白表达能通过活化p53通路促进细胞凋亡,并增加肝癌细胞对化疗药物的敏感性。
