RBM5、EGFR和KRAS在非小细胞肺癌中的表达及其意义
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摘要
目的检测非小细胞肺癌组织RBM5、EGFR及KRAS表达水平,并进一步分析上述三种基因表达与非小细胞肺癌患者临床、病理相关性,为肺癌早期诊断及分子靶向治疗提供依据。方法收集2017年8月至2018年8月在吉林大学第二医院接受手术的42例NSCLC患者癌组织及癌旁组织标本,采用Western blot及实时荧光定量PCR(RT-qPCR)方法检测RBM5、EGFR和KRAS蛋白及mRNA相对表达水平,同时采用ELISA方法检测44例患者外周血清中上述基因表达水平,并进一步分析其临床病理相关性。结果在组织学分析中,与癌旁组织比较,肺癌组织中RBM5mRNA表达水平降低(P<0.05),降低水平与吸烟史、淋巴结转移等相关;EGFR mRNA表达水平升高(P<0.05),其中伴有淋巴结转移组中EGFR相对表达量高于无淋巴结转移组(P<0.05);KRAS mRNA表达水平增高与临床分期、淋巴结转移等相关。在血清学ELISA分析中,与健康人比较,肺癌患者血清中RBM5表达水平降低(P<0.01),其中伴有淋巴结转移组中RBM5表达量低于无淋巴结转移组(P<0.05);EGFR和KRAS表达水平升高(P<0.01),表达水平的升高与肺癌患者临床分期及淋巴结转移无显著相关性。结论肿瘤抑制基因RBM5在NSCLC发生发展过程中起到负性调控作用,而EGFR和KRAS起到正性调控作用;血清中RBM5和EGFR可能成为判断肺癌早期诊断及预测转移的新指标,提供肺癌治疗新靶点。
Objective The aim of the study is to detect the protein and mRNA expression level of RBM5,EGFR and KRAS in serum and tissues in patients with NSCLC,to analyze the correlation between the expression level and clinical pathological parameters.Accordingly,it can be helpful to provide clues for early diagnosis of lung cancer and targeted treatment.Methods The fresh postoperative specimens of 42 cases of carcinoma tissue and para-carcinoma tissue from August 2017 to August 2018 at the Second Hospital of Jilin University were selected.The protein and mRNA relative expression level of RBM5、EGFR and KRAS in tissues were measured by Western blot and the real-time fluorescent quantitative PCR(RT-qPCR).And the protein expression level of the above genes in serum of 44 patients were detected by ELISA.Results Compared to para-cancerous tissues,the relative expression of RBM5 mRNA was significantly decreased in NSCLC tissues(P<0.05),and the decrease was related to smoking history and lymph node metastasis;the expression of EGFR mRNA was significantly increased in NSCLC tissues(P<0.05),and the relative expression of EGFR in the lymph node metastasis group was higher than that in the non-lymph node metastasis group(P<0.05).The expression level of serum RBM5 protein of the patients in NSCLC was significantly lower than that in controls(P<0.01),and the extent of decrease was related to the status of smoking and metastasis.Increased expression of KRAS mRNA is associated with clinical stage,lymph node metastasis,etc.The expression of RBM5,EGFR and KRAS mRNA in NSCLC tissues and para-cancerous tissues was related to some clinical pathological factors.The expression level of EGFR and KRAS protein in patients' serum were significantly higher than those in healthy control group(P<0.001).Conclusion The tumor suppressor gene RBM5 plays a negative regulatory role in the development of NSCLC,while EGFR and KRAS play apositive regulatory role;serum RBM5 and EGFR may become new indicators for early diagnosis and predictive metastasis of lung cancer,providing new treatment for lung cancer.
Objective The aim of the study is to detect the protein and mRNA expression level of RBM5,EGFR and KRAS in serum and tissues in patients with NSCLC,to analyze the correlation between the expression level and clinical pathological parameters.Accordingly,it can be helpful to provide clues for early diagnosis of lung cancer and targeted treatment.Methods The fresh postoperative specimens of 42 cases of carcinoma tissue and para-carcinoma tissue from August 2017 to August 2018 at the Second Hospital of Jilin University were selected.The protein and mRNA relative expression level of RBM5、EGFR and KRAS in tissues were measured by Western blot and the real-time fluorescent quantitative PCR(RT-qPCR).And the protein expression level of the above genes in serum of 44 patients were detected by ELISA.Results Compared to para-cancerous tissues,the relative expression of RBM5 mRNA was significantly decreased in NSCLC tissues(P<0.05),and the decrease was related to smoking history and lymph node metastasis;the expression of EGFR mRNA was significantly increased in NSCLC tissues(P<0.05),and the relative expression of EGFR in the lymph node metastasis group was higher than that in the non-lymph node metastasis group(P<0.05).The expression level of serum RBM5 protein of the patients in NSCLC was significantly lower than that in controls(P<0.01),and the extent of decrease was related to the status of smoking and metastasis.Increased expression of KRAS mRNA is associated with clinical stage,lymph node metastasis,etc.The expression of RBM5,EGFR and KRAS mRNA in NSCLC tissues and para-cancerous tissues was related to some clinical pathological factors.The expression level of EGFR and KRAS protein in patients' serum were significantly higher than those in healthy control group(P<0.001).Conclusion The tumor suppressor gene RBM5 plays a negative regulatory role in the development of NSCLC,while EGFR and KRAS play apositive regulatory role;serum RBM5 and EGFR may become new indicators for early diagnosis and predictive metastasis of lung cancer,providing new treatment for lung cancer.
引文
[1]Bray F,Ferlay J,Soerjomataram I,et al.Global cancer statistics2018:GLOBOCAN estimates of incidence and mortality worldwide for 36cancers in 185countries[J].Ca-Cancer J Clin,2018,68:394.
[2]McIntyre A,Ganti AK.Lung cancer-A global perspective[J].J Surg Oncol,2017,115:550.
[3]Wang H,Zeng C,Li X,Wang Y,et al.Cost-utility of afatinib and gefitinib as first-line treatment for EGFR-mutated advanced nonsmall-cell lung cancer[J].Future oncology,2018.
[4]Ettinger DS,Wood DE,Akerley W,et al.Non-Small Cell Lung Cancer,Version 6.2015[J].Journal of the National Comprehensive Cancer Network:JNCCN,2015,13:515.
[5]Wei MH,Latif F,Bader S,et al.Construction of a 600-kilobase cosmid clone contig and generation of a transcriptional map surrounding the lung cancer tumor suppressor gene(TSG)locus on human chromosome 3p21.3:progress toward the isolation of a lung cancer TSG[J].Cancer research,1996,56:1487.
[6]Oh JJ,Razfar A,Delgado I,et al.3p21.3tumor suppressor gene H37/Luca15/RBM5inhibits growth of human lung cancer cells through cell cycle arrest and apoptosis[J].Cancer research,2006,66:3419.
[7]Ji L,Minna JD,Roth JA.3p21.3tumor suppressor cluster:prospects for translational applications[J].Future oncology,2005,1:79.
[8]Sutherland LC,Wang K,Robinson AG.RBM5as a putative tumor suppressor gene for lung cancer[J].Journal of thoracic oncology:official publication of the International Association for the Study of Lung Cancer,2010,5:294.
[9]Ciardiello F,Tortora G.EGFR antagonists in cancer treatment[J].The New England journal of medicine,2008,358:1160.
[10]Hirsch FR,Varella-Garcia M,Cappuzzo F.Predictive value of EGFR and HER2 overexpression in advanced non-small-cell lung cancer[J].Oncogene,2009,28Suppl 1:S32.
[11]Bae NC,Chae MH,Lee MH,et al.EGFR,ERBB2,and KRAS mutations in Korean non-small cell lung cancer patients[J].Cancer genetics and cytogenetics,2007,173:107.
[12]Liang H,Zhang J,Shao C,et al.Differential expression of RBM5,EGFR and KRAS mRNA and protein in non-small cell lung cancer tissues[J].Journal of experimental&clinical cancer research:CR,2012,31:36.
[13]Detterbeck FC,Chansky K,Groome P,et al.The IASLC Lung Cancer Staging Project:Methodology and Validation Used in the Development of Proposals for Revision of the Stage Classification of NSCLC in the Forthcoming(Eighth)Edition of the TNM Classification of Lung Cancer[J].Journal of Thoracic Oncology,2016,11:1433.
[14]Oh JJ,West AR,Fishbein MC,et al.A candidate tumor suppressor gene,H37,from the human lung cancer tumor suppressor locus 3p21.3[J].Cancer research,2002,62:3207.
[15]Welling DB,Lasak JM,Akhmametyeva E,et al.cDNA microarray analysis of vestibular schwannomas[J].Otology&neurotology:official publication of the American Otological Society,American Neurotology Society[and]European Academy of Otology and Neurotology,2002,23:736.
[16]Gately K,Forde L,Cuffe S,et al.High coexpression of both EGFR and IGF1Rcorrelates with poor patient prognosis in resected non-small-cell lung cancer[J].Clin Lung Cancer,2014,15:58.
[17]Westcott PM,To MD.The genetics and biology of KRAS in lung cancer[J].Chinese Journal of Cancer,2013,32:63.
[18]Amos CI,Xu W,Spitz MR.Is there a genetic basis for lung cancer susceptibility[J]? Recent results in cancer research Fortschritte der Krebsforschung Progres dans les recherches sur le cancer,1999,151:3.
[19]Oh JJ,Koegel AK,Phan DT,et al.The two single nucleotide polymorphisms in the H37/RBM5tumour suppressor gene at3p21.3correlated with different subtypes of non-small cell lung cancers[J].Lung Cancer,2007,58:7.
[20]Dogan S,Shen R,Ang DC,et al.Molecular epidemiology of EGFR and KRAS mutations in 3,026lung adenocarcinomas:higher susceptibility of women to smoking-related KRAS-mutant cancers[J].Clin Cancer Res,2012,18:6169.
[21]Herbst RS,Shin DM.Monoclonal antibodies to target epidermal growth factor receptor-positive tumors:a new paradigm for cancer therapy[J].Cancer,2002,94:1593.
[22]Haghgoo SM,Khosravi A,Mortaz E,et al.Prognostic value of rare and complex mutations in EGFR and serum levels of soluble EGFR and its ligands in non-small cell lung carcinoma patients[J].Clinical biochemistry,2017,50:293.
[23]Ryan BM,Lefort F,McManus R,et al.A prospective study of circulating mutant KRAS2in the serum of patients with colorectal neoplasia:strong prognostic indicator in postoperative follow up[J].Gut,2003,52:101.
[24]Peng J,Valeshabad AK,Li Q,Wang Y.Differential expression of RBM5and KRAS in pancreatic ductal adenocarcinoma and their association with clinicopathological features[J].Oncology letters,2013,5:1000.
[2]McIntyre A,Ganti AK.Lung cancer-A global perspective[J].J Surg Oncol,2017,115:550.
[3]Wang H,Zeng C,Li X,Wang Y,et al.Cost-utility of afatinib and gefitinib as first-line treatment for EGFR-mutated advanced nonsmall-cell lung cancer[J].Future oncology,2018.
[4]Ettinger DS,Wood DE,Akerley W,et al.Non-Small Cell Lung Cancer,Version 6.2015[J].Journal of the National Comprehensive Cancer Network:JNCCN,2015,13:515.
[5]Wei MH,Latif F,Bader S,et al.Construction of a 600-kilobase cosmid clone contig and generation of a transcriptional map surrounding the lung cancer tumor suppressor gene(TSG)locus on human chromosome 3p21.3:progress toward the isolation of a lung cancer TSG[J].Cancer research,1996,56:1487.
[6]Oh JJ,Razfar A,Delgado I,et al.3p21.3tumor suppressor gene H37/Luca15/RBM5inhibits growth of human lung cancer cells through cell cycle arrest and apoptosis[J].Cancer research,2006,66:3419.
[7]Ji L,Minna JD,Roth JA.3p21.3tumor suppressor cluster:prospects for translational applications[J].Future oncology,2005,1:79.
[8]Sutherland LC,Wang K,Robinson AG.RBM5as a putative tumor suppressor gene for lung cancer[J].Journal of thoracic oncology:official publication of the International Association for the Study of Lung Cancer,2010,5:294.
[9]Ciardiello F,Tortora G.EGFR antagonists in cancer treatment[J].The New England journal of medicine,2008,358:1160.
[10]Hirsch FR,Varella-Garcia M,Cappuzzo F.Predictive value of EGFR and HER2 overexpression in advanced non-small-cell lung cancer[J].Oncogene,2009,28Suppl 1:S32.
[11]Bae NC,Chae MH,Lee MH,et al.EGFR,ERBB2,and KRAS mutations in Korean non-small cell lung cancer patients[J].Cancer genetics and cytogenetics,2007,173:107.
[12]Liang H,Zhang J,Shao C,et al.Differential expression of RBM5,EGFR and KRAS mRNA and protein in non-small cell lung cancer tissues[J].Journal of experimental&clinical cancer research:CR,2012,31:36.
[13]Detterbeck FC,Chansky K,Groome P,et al.The IASLC Lung Cancer Staging Project:Methodology and Validation Used in the Development of Proposals for Revision of the Stage Classification of NSCLC in the Forthcoming(Eighth)Edition of the TNM Classification of Lung Cancer[J].Journal of Thoracic Oncology,2016,11:1433.
[14]Oh JJ,West AR,Fishbein MC,et al.A candidate tumor suppressor gene,H37,from the human lung cancer tumor suppressor locus 3p21.3[J].Cancer research,2002,62:3207.
[15]Welling DB,Lasak JM,Akhmametyeva E,et al.cDNA microarray analysis of vestibular schwannomas[J].Otology&neurotology:official publication of the American Otological Society,American Neurotology Society[and]European Academy of Otology and Neurotology,2002,23:736.
[16]Gately K,Forde L,Cuffe S,et al.High coexpression of both EGFR and IGF1Rcorrelates with poor patient prognosis in resected non-small-cell lung cancer[J].Clin Lung Cancer,2014,15:58.
[17]Westcott PM,To MD.The genetics and biology of KRAS in lung cancer[J].Chinese Journal of Cancer,2013,32:63.
[18]Amos CI,Xu W,Spitz MR.Is there a genetic basis for lung cancer susceptibility[J]? Recent results in cancer research Fortschritte der Krebsforschung Progres dans les recherches sur le cancer,1999,151:3.
[19]Oh JJ,Koegel AK,Phan DT,et al.The two single nucleotide polymorphisms in the H37/RBM5tumour suppressor gene at3p21.3correlated with different subtypes of non-small cell lung cancers[J].Lung Cancer,2007,58:7.
[20]Dogan S,Shen R,Ang DC,et al.Molecular epidemiology of EGFR and KRAS mutations in 3,026lung adenocarcinomas:higher susceptibility of women to smoking-related KRAS-mutant cancers[J].Clin Cancer Res,2012,18:6169.
[21]Herbst RS,Shin DM.Monoclonal antibodies to target epidermal growth factor receptor-positive tumors:a new paradigm for cancer therapy[J].Cancer,2002,94:1593.
[22]Haghgoo SM,Khosravi A,Mortaz E,et al.Prognostic value of rare and complex mutations in EGFR and serum levels of soluble EGFR and its ligands in non-small cell lung carcinoma patients[J].Clinical biochemistry,2017,50:293.
[23]Ryan BM,Lefort F,McManus R,et al.A prospective study of circulating mutant KRAS2in the serum of patients with colorectal neoplasia:strong prognostic indicator in postoperative follow up[J].Gut,2003,52:101.
[24]Peng J,Valeshabad AK,Li Q,Wang Y.Differential expression of RBM5and KRAS in pancreatic ductal adenocarcinoma and their association with clinicopathological features[J].Oncology letters,2013,5:1000.