BACKGROUND The objectives of this study were to identify hub genes and biological pathways involved in lung adenocarcinoma(LUAD)via bioinformatics analysis,and investigate potential therapeutic targets.AIM To determin...BACKGROUND The objectives of this study were to identify hub genes and biological pathways involved in lung adenocarcinoma(LUAD)via bioinformatics analysis,and investigate potential therapeutic targets.AIM To determine reliable prognostic biomarkers for early diagnosis and treatment of LUAD.METHODS To identify potential therapeutic targets for LUAD,two microarray datasets derived from the Gene Expression Omnibus(GEO)database were analyzed,GSE3116959 and GSE118370.Differentially expressed genes(DEGs)in LUAD and normal tissues were identified using the GEO2R tool.The Hiplot database was then used to generate a volcanic map of the DEGs.Weighted gene co-expression network analysis was conducted to cluster the genes in GSE116959 and GSE-118370 into different modules,and identify immune genes shared between them.A protein-protein interaction network was established using the Search Tool for the Retrieval of Interacting Genes database,then the CytoNCA and CytoHubba components of Cytoscape software were used to visualize the genes.Hub genes with high scores and co-expression were identified,and the Database for Annotation,Visualization and Integrated Discovery was used to perform enrichment analysis of these genes.The diagnostic and prognostic values of the hub genes were calculated using receiver operating characteristic curves and Kaplan-Meier survival analysis,and gene-set enrichment analysis was conducted.The University of Alabama at Birmingham Cancer data analysis portal was used to analyze relationships between the hub genes and normal specimens,as well as their expression during tumor progression.Lastly,validation of protein expression was conducted on the identified hub genes via the Human Protein Atlas database.RESULTS Three hub genes with high connectivity were identified;cellular retinoic acid binding protein 2(CRABP2),matrix metallopeptidase 12(MMP12),and DNA topoisomerase II alpha(TOP2A).High expression of these genes was associated with a poor LUAD prognosis,and the genes exhibited high diagnostic value.CONCLUSION Expression levels of CRABP2,MMP12,and TOP2A in LUAD were higher than those in normal lung tissue.This observation has diagnostic value,and is linked to poor LUAD prognosis.These genes may be biomarkers and therapeutic targets in LUAD,but further research is warranted to investigate their usefulness in these respects.展开更多
目的通过脂多糖(Lipopolysaccharide,LPS)诱导的脓毒症模型验证G蛋白偶联受体120(G-protein coupled receptor120,GPR120)基因对NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor thermal protein domain associated protein 3,NLRP3...目的通过脂多糖(Lipopolysaccharide,LPS)诱导的脓毒症模型验证G蛋白偶联受体120(G-protein coupled receptor120,GPR120)基因对NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)炎症小体及肺损伤的影响,并探索其调控分子机制。方法通过C57BL/6小鼠构建体内脓毒症模型,通过GPR120基因激动剂TUG891进行干预,验证GPR120基因对脓毒症小鼠肺损伤的保护作用;然后进行转录组测序,筛选差异信号通路,并在动物模型中验证NLRP3炎症小体及调控蛋白的差异表达。通过慢病毒转染构建GPR120基因过表达/低表达的Raw264.7单核巨噬细胞株,观察GPR120基因对NLRP3炎症小体的调控作用。结果与脓毒症组相比,LPS+TUG891组小鼠肺组织中包括cAMP通路基因在内的77个基因表达显著上调,37个基因表达下降。LPS组的GPR120水平较正常对照组显著降低,同时cAMP/PKA信号通路关键蛋白CREB及PKA表达减少,NLRP3、Caspase-1及IL-1β等炎症小体激活相关蛋白水平升高(P<0.01),予以TUG891处理后,组织内GPR120表达回升,cAMP/PKA信号通路重新被激活(P<0.01),NLRP3炎症小体蛋白活化程度下降(P<0.05)。体外实验中,LPS诱导的脓毒症可引起细胞增殖活性下降,GPR120基因在脓毒症巨噬细胞中表达减低(P<0.001),通过干预GPR120基因表达,证实GPR120基因可负性调控NLRP3炎症小体的活化程度及细胞炎症反应(P<0.01)。结论脓毒症中GPR120基因的激活可通过抑制NLRP3炎症小体的活化,减轻脓毒症的炎症反应及肺损伤。展开更多
Objective: To study the expressions of two isoforms of Livin in tissues of lung cancer and their relations to histological types and chemotherapy, and to study their correlations to the expression of caspase-3 as well...Objective: To study the expressions of two isoforms of Livin in tissues of lung cancer and their relations to histological types and chemotherapy, and to study their correlations to the expression of caspase-3 as well. Methods: Ex-pressions of Livin isoforms α, β and caspase-3 were detected by reverse transcription polymerase chain reaction (RT-PCR) assay in lung cancer tissues as well as in controls. Results: Livin isoforms α and β were expressed in 12 of 27, and 19 of 27 lung cancer tissues respectively, much more than those in lung para-cancerous [both were (0/6)] or benign disease lung tissues (0/12, 1/12; P < 0.01 and P < 0.01). Moreover, they were detected in 7/14, 9 /14 lung adenocarcinomas and 4/12, 9/12 squamocellular and large cell carcinomas, respectively, and both showed expressions in one small cell carcinoma. The levels of these two isoforms in lung cancer were significantly higher than those in controls by Gel imaging system (P < 0.05 and P < 0.05), the former was higher in adenocarcinoma than that in squamocellular carcinoma (P < 0.05), while the latter was the same in both (P > 0.05). Meanwhile, the levels of caspase-3 in lung cancer were significantly lower than those in controls, and it was suggested to be negatively associated with either each of two isoforms or their sum (P < 0.05, P < 0.01 and P < 0.01). Two isoforms of Livin expression seemed to increase after chemotherapy but not related to clinical stages (P > 0.05). Conclusion: Two isoforms of Livin are differently expressed in different histological types of lung cancer and may contribute to corresponding cancerous development; the levels of Livin are negatively associated with those of caspase-3, this may be due to the fact that Livin could resist against apoptosis; high expression of Livin seems to be related to chemotherapy but not clinical stages.展开更多
Objective: To observe the efficacy and safety of cetuximab combined with chemotherapy in advanced non-small-cell lung cancer (NSCLC), and to investigate the association of status of K-RAS gene mutation and epiderma...Objective: To observe the efficacy and safety of cetuximab combined with chemotherapy in advanced non-small-cell lung cancer (NSCLC), and to investigate the association of status of K-RAS gene mutation and epidermal growth factor receptor (EGFR) genotype with clinical outcome. Methods: Between Jan. 2006 and Sep. 2009, nineteen patients with advanced NSCLC received cetuximab (〉4 weeks) combined with chemotherapy in Department of Thoracic Oncology at Beijing Cancer Hospital. Response, survival and toxicity were retrospectively assessed, epidermal growth factor receptor (EGFR) protein expression was evaluated by ELISA Kit. The status of K-RAS gene mutation was tested by PCR-RFLP and EGFR gene amplification was measured by EGFR fluorescence in situ hybridization (FISH). Results: Partial response(PR) was observed in 26.3%(5/19) of the patients and stable disease(SD) in 52.6%(10/19). Median progression free survival(PFS) was 6 months (95% CI: 3.6-8.4). Median overall survival (MST) and 1-year survival rate(SR) were 10.6 months (95% CI: 6.6-14.6) and 47.6%, respectively. Mild or moderate skin rash was the most common toxicity related with cetuximab. K-RAS gene mutation, EGFR protein level and amplification have little correlation with prognosis. Conclusion: Cetuximab combined with chemotherapy was tolerable and the skin rash related with cetuximab was mild to moderate. Cetuximab may prolong survival of the patients who failed to previous chemotherapy.展开更多
Antigen containing, allogeneic cells secreting the genetically modified protein and peptide-chaperone gp96-Ig cross, prime and expand antigen specific CD8 T cells with therapeutic antitumor activity in mice. In a firs...Antigen containing, allogeneic cells secreting the genetically modified protein and peptide-chaperone gp96-Ig cross, prime and expand antigen specific CD8 T cells with therapeutic antitumor activity in mice. In a first in man phase I study, we now report the results of therapeutic vaccination of non-small cell lung cancer (NSCLC) patients with an established, allogeneic non-small cell lung adenocarcinoma cell line secreting gp96-Ig. Advanced NSCLC-patients stage IIIB or IV of any histological subtype were enrolled and treated with up to 36 vaccinations over the course of 18 weeks. Primary endpoint was safety, secondary endpoints tumor response and overall survival. Measurement of tumor antigen specific CD8 CTL responses is precluded by the lack of known NSCLC associated antigens. Therefore, we measured patient CD8 T cell-IFN-γ responses to allo-antigens of the vaccine cells as surrogate for tumor antigen specific CD8 CTL. In 7 of 18 treated patients tumor growth was stabilized, however none of the 18 patients had an objective tumor response by RECIST criteria. Of 15 patients evaluable for immune response, 11 responded to vaccination with more than twofold increase in CD8-IFN-γ frequency above baseline. These patients had a median survival time of 16.5 months. Four patients who had no CD8 response above base line had survival times from 2.1 to 6.7 months. Our data are consistent with the concept that generation of CD8 CTL by therapeutic vaccination may delay tumor growth and progression and mediate prolonged survival even in the absence of objective tumor responses. Further studies will be required to test this concept and promising result.展开更多
Objective:To investigate the effects of Intron on the EMT capability of non-small cell lung cancer cell line PC-9.Methods:Firstly,using the psiCHECK-2 plasmid as a basic framework to construct the recombinant plasmid ...Objective:To investigate the effects of Intron on the EMT capability of non-small cell lung cancer cell line PC-9.Methods:Firstly,using the psiCHECK-2 plasmid as a basic framework to construct the recombinant plasmid of psiCHECK-2-Intron dual-luciferase reporter gene;secondly,the psiCHECK-2-Intron and psiCHECK-2 were transfected into PC-9 cells respectively.The migration and invasion abilities of PC-9 cells were analyzed by Matrigel assay.The expression changes of EMT related hallmarks,including N-cadherin,β-catenin and snail,were detected by qRT-PCR and Western Blotting.Results:Compared with the control group,the migration and invasion abilities of PC-9 cells in Intron group significantly decreased(p<0.001).The expression of N-cadherin,β-catenin and snail also down-regulated(p<0.001).Conclusion:The introns could inhibit the EMT of PC-9 cells.展开更多
背景与目的:可变剪接在细胞增殖、生长、凋亡及分化等复杂的蛋白质功能调控中发挥着至关重要的作用,能参与包括肿瘤在内的多种疾病的发生、发展过程,目前大多数可变剪接调控因子的功能及其作用机制尚不清楚。本研究旨在探讨KH型剪接调...背景与目的:可变剪接在细胞增殖、生长、凋亡及分化等复杂的蛋白质功能调控中发挥着至关重要的作用,能参与包括肿瘤在内的多种疾病的发生、发展过程,目前大多数可变剪接调控因子的功能及其作用机制尚不清楚。本研究旨在探讨KH型剪接调控蛋白(KH-type splicing regulatory protein,KHSRP)在肺腺癌中的差异剪接靶基因及靶基因的表达与患者预后及免疫细胞功能的相关性。方法:通过高通量可变剪接测序,筛选KHSRP下游差异剪接靶基因。通过肿瘤免疫评估资源(Tumor IMmune Estimation Resource,TIMER)和阿拉巴马大学伯明翰分校癌症数据分析门户(the University of Alabama at Birmingham cancer data analysis portal,ULCAN)分析KHSRP下游差异剪接靶基因在肺腺癌中的表达。通过Kaplan数据库分析KHSRP下游差异剪接靶基因的表达与患者预后的相关性。通过TIMER免疫模块分析KHSRP下游差异剪接靶基因与免疫细胞功能的相关性。结果:NUMB、ADD3及LIMCH1为KHSRP的下游关键可变剪接靶基因。NUMB、ADD3及LIMCH1在肺腺癌肿瘤组织中均呈低表达,其低表达者预后不良。NUMB、ADD3及LIMCH1与免疫细胞功能呈正相关。结论:在肺腺癌中,NUMB、ADD3及LIMCH1为KHSRP的差异剪接靶基因,且NUMB、ADD3及LIMCH1低表达会导致预后不良,并且与免疫细胞功能呈正相关。展开更多
文摘BACKGROUND The objectives of this study were to identify hub genes and biological pathways involved in lung adenocarcinoma(LUAD)via bioinformatics analysis,and investigate potential therapeutic targets.AIM To determine reliable prognostic biomarkers for early diagnosis and treatment of LUAD.METHODS To identify potential therapeutic targets for LUAD,two microarray datasets derived from the Gene Expression Omnibus(GEO)database were analyzed,GSE3116959 and GSE118370.Differentially expressed genes(DEGs)in LUAD and normal tissues were identified using the GEO2R tool.The Hiplot database was then used to generate a volcanic map of the DEGs.Weighted gene co-expression network analysis was conducted to cluster the genes in GSE116959 and GSE-118370 into different modules,and identify immune genes shared between them.A protein-protein interaction network was established using the Search Tool for the Retrieval of Interacting Genes database,then the CytoNCA and CytoHubba components of Cytoscape software were used to visualize the genes.Hub genes with high scores and co-expression were identified,and the Database for Annotation,Visualization and Integrated Discovery was used to perform enrichment analysis of these genes.The diagnostic and prognostic values of the hub genes were calculated using receiver operating characteristic curves and Kaplan-Meier survival analysis,and gene-set enrichment analysis was conducted.The University of Alabama at Birmingham Cancer data analysis portal was used to analyze relationships between the hub genes and normal specimens,as well as their expression during tumor progression.Lastly,validation of protein expression was conducted on the identified hub genes via the Human Protein Atlas database.RESULTS Three hub genes with high connectivity were identified;cellular retinoic acid binding protein 2(CRABP2),matrix metallopeptidase 12(MMP12),and DNA topoisomerase II alpha(TOP2A).High expression of these genes was associated with a poor LUAD prognosis,and the genes exhibited high diagnostic value.CONCLUSION Expression levels of CRABP2,MMP12,and TOP2A in LUAD were higher than those in normal lung tissue.This observation has diagnostic value,and is linked to poor LUAD prognosis.These genes may be biomarkers and therapeutic targets in LUAD,but further research is warranted to investigate their usefulness in these respects.
文摘目的通过脂多糖(Lipopolysaccharide,LPS)诱导的脓毒症模型验证G蛋白偶联受体120(G-protein coupled receptor120,GPR120)基因对NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)炎症小体及肺损伤的影响,并探索其调控分子机制。方法通过C57BL/6小鼠构建体内脓毒症模型,通过GPR120基因激动剂TUG891进行干预,验证GPR120基因对脓毒症小鼠肺损伤的保护作用;然后进行转录组测序,筛选差异信号通路,并在动物模型中验证NLRP3炎症小体及调控蛋白的差异表达。通过慢病毒转染构建GPR120基因过表达/低表达的Raw264.7单核巨噬细胞株,观察GPR120基因对NLRP3炎症小体的调控作用。结果与脓毒症组相比,LPS+TUG891组小鼠肺组织中包括cAMP通路基因在内的77个基因表达显著上调,37个基因表达下降。LPS组的GPR120水平较正常对照组显著降低,同时cAMP/PKA信号通路关键蛋白CREB及PKA表达减少,NLRP3、Caspase-1及IL-1β等炎症小体激活相关蛋白水平升高(P<0.01),予以TUG891处理后,组织内GPR120表达回升,cAMP/PKA信号通路重新被激活(P<0.01),NLRP3炎症小体蛋白活化程度下降(P<0.05)。体外实验中,LPS诱导的脓毒症可引起细胞增殖活性下降,GPR120基因在脓毒症巨噬细胞中表达减低(P<0.001),通过干预GPR120基因表达,证实GPR120基因可负性调控NLRP3炎症小体的活化程度及细胞炎症反应(P<0.01)。结论脓毒症中GPR120基因的激活可通过抑制NLRP3炎症小体的活化,减轻脓毒症的炎症反应及肺损伤。
文摘Objective: To study the expressions of two isoforms of Livin in tissues of lung cancer and their relations to histological types and chemotherapy, and to study their correlations to the expression of caspase-3 as well. Methods: Ex-pressions of Livin isoforms α, β and caspase-3 were detected by reverse transcription polymerase chain reaction (RT-PCR) assay in lung cancer tissues as well as in controls. Results: Livin isoforms α and β were expressed in 12 of 27, and 19 of 27 lung cancer tissues respectively, much more than those in lung para-cancerous [both were (0/6)] or benign disease lung tissues (0/12, 1/12; P < 0.01 and P < 0.01). Moreover, they were detected in 7/14, 9 /14 lung adenocarcinomas and 4/12, 9/12 squamocellular and large cell carcinomas, respectively, and both showed expressions in one small cell carcinoma. The levels of these two isoforms in lung cancer were significantly higher than those in controls by Gel imaging system (P < 0.05 and P < 0.05), the former was higher in adenocarcinoma than that in squamocellular carcinoma (P < 0.05), while the latter was the same in both (P > 0.05). Meanwhile, the levels of caspase-3 in lung cancer were significantly lower than those in controls, and it was suggested to be negatively associated with either each of two isoforms or their sum (P < 0.05, P < 0.01 and P < 0.01). Two isoforms of Livin expression seemed to increase after chemotherapy but not related to clinical stages (P > 0.05). Conclusion: Two isoforms of Livin are differently expressed in different histological types of lung cancer and may contribute to corresponding cancerous development; the levels of Livin are negatively associated with those of caspase-3, this may be due to the fact that Livin could resist against apoptosis; high expression of Livin seems to be related to chemotherapy but not clinical stages.
文摘Objective: To observe the efficacy and safety of cetuximab combined with chemotherapy in advanced non-small-cell lung cancer (NSCLC), and to investigate the association of status of K-RAS gene mutation and epidermal growth factor receptor (EGFR) genotype with clinical outcome. Methods: Between Jan. 2006 and Sep. 2009, nineteen patients with advanced NSCLC received cetuximab (〉4 weeks) combined with chemotherapy in Department of Thoracic Oncology at Beijing Cancer Hospital. Response, survival and toxicity were retrospectively assessed, epidermal growth factor receptor (EGFR) protein expression was evaluated by ELISA Kit. The status of K-RAS gene mutation was tested by PCR-RFLP and EGFR gene amplification was measured by EGFR fluorescence in situ hybridization (FISH). Results: Partial response(PR) was observed in 26.3%(5/19) of the patients and stable disease(SD) in 52.6%(10/19). Median progression free survival(PFS) was 6 months (95% CI: 3.6-8.4). Median overall survival (MST) and 1-year survival rate(SR) were 10.6 months (95% CI: 6.6-14.6) and 47.6%, respectively. Mild or moderate skin rash was the most common toxicity related with cetuximab. K-RAS gene mutation, EGFR protein level and amplification have little correlation with prognosis. Conclusion: Cetuximab combined with chemotherapy was tolerable and the skin rash related with cetuximab was mild to moderate. Cetuximab may prolong survival of the patients who failed to previous chemotherapy.
文摘Antigen containing, allogeneic cells secreting the genetically modified protein and peptide-chaperone gp96-Ig cross, prime and expand antigen specific CD8 T cells with therapeutic antitumor activity in mice. In a first in man phase I study, we now report the results of therapeutic vaccination of non-small cell lung cancer (NSCLC) patients with an established, allogeneic non-small cell lung adenocarcinoma cell line secreting gp96-Ig. Advanced NSCLC-patients stage IIIB or IV of any histological subtype were enrolled and treated with up to 36 vaccinations over the course of 18 weeks. Primary endpoint was safety, secondary endpoints tumor response and overall survival. Measurement of tumor antigen specific CD8 CTL responses is precluded by the lack of known NSCLC associated antigens. Therefore, we measured patient CD8 T cell-IFN-γ responses to allo-antigens of the vaccine cells as surrogate for tumor antigen specific CD8 CTL. In 7 of 18 treated patients tumor growth was stabilized, however none of the 18 patients had an objective tumor response by RECIST criteria. Of 15 patients evaluable for immune response, 11 responded to vaccination with more than twofold increase in CD8-IFN-γ frequency above baseline. These patients had a median survival time of 16.5 months. Four patients who had no CD8 response above base line had survival times from 2.1 to 6.7 months. Our data are consistent with the concept that generation of CD8 CTL by therapeutic vaccination may delay tumor growth and progression and mediate prolonged survival even in the absence of objective tumor responses. Further studies will be required to test this concept and promising result.
基金National Natural Science Foundation of China(No.81660474)。
文摘Objective:To investigate the effects of Intron on the EMT capability of non-small cell lung cancer cell line PC-9.Methods:Firstly,using the psiCHECK-2 plasmid as a basic framework to construct the recombinant plasmid of psiCHECK-2-Intron dual-luciferase reporter gene;secondly,the psiCHECK-2-Intron and psiCHECK-2 were transfected into PC-9 cells respectively.The migration and invasion abilities of PC-9 cells were analyzed by Matrigel assay.The expression changes of EMT related hallmarks,including N-cadherin,β-catenin and snail,were detected by qRT-PCR and Western Blotting.Results:Compared with the control group,the migration and invasion abilities of PC-9 cells in Intron group significantly decreased(p<0.001).The expression of N-cadherin,β-catenin and snail also down-regulated(p<0.001).Conclusion:The introns could inhibit the EMT of PC-9 cells.
文摘背景与目的:可变剪接在细胞增殖、生长、凋亡及分化等复杂的蛋白质功能调控中发挥着至关重要的作用,能参与包括肿瘤在内的多种疾病的发生、发展过程,目前大多数可变剪接调控因子的功能及其作用机制尚不清楚。本研究旨在探讨KH型剪接调控蛋白(KH-type splicing regulatory protein,KHSRP)在肺腺癌中的差异剪接靶基因及靶基因的表达与患者预后及免疫细胞功能的相关性。方法:通过高通量可变剪接测序,筛选KHSRP下游差异剪接靶基因。通过肿瘤免疫评估资源(Tumor IMmune Estimation Resource,TIMER)和阿拉巴马大学伯明翰分校癌症数据分析门户(the University of Alabama at Birmingham cancer data analysis portal,ULCAN)分析KHSRP下游差异剪接靶基因在肺腺癌中的表达。通过Kaplan数据库分析KHSRP下游差异剪接靶基因的表达与患者预后的相关性。通过TIMER免疫模块分析KHSRP下游差异剪接靶基因与免疫细胞功能的相关性。结果:NUMB、ADD3及LIMCH1为KHSRP的下游关键可变剪接靶基因。NUMB、ADD3及LIMCH1在肺腺癌肿瘤组织中均呈低表达,其低表达者预后不良。NUMB、ADD3及LIMCH1与免疫细胞功能呈正相关。结论:在肺腺癌中,NUMB、ADD3及LIMCH1为KHSRP的差异剪接靶基因,且NUMB、ADD3及LIMCH1低表达会导致预后不良,并且与免疫细胞功能呈正相关。