乏氧和低糖条件下低表达ESE3促进胰腺癌细胞的侵袭
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摘要
目的观察乏氧和低糖条件下胰腺癌细胞中上皮特异性ETS转录因子3(ESE3)表达的变化及其对胰腺癌细胞侵袭能力的影响。方法收集在天津医科大学肿瘤医院2005年1月—2014年10月期间行根治性手术治疗的99例胰腺癌患者的胰腺癌组织和16例邻近正常胰腺组织标本。采用免疫组化法分析癌组织和邻近正常胰腺组织中ESE3的表达水平,探究其表达水平与胰腺癌患者生存预后间的关系;进一步在TCGA数据库中分析ESE3的mRNA表达水平与147例胰腺癌患者的临床病理学特征间的关系;构建胰腺癌细胞PANC-1过表达ESE3稳定转染细胞系并验证;Transwell法探究过表达ESE3的胰腺癌细胞侵袭能力的变化;利用蛋白质免疫印迹法检测乏氧和低糖条件下胰腺癌细胞中乏氧诱导因子1α(HIF-1α)、葡萄糖调节蛋白78(GRP78)和ESE3蛋白表达水平的变化。结果胰腺癌组织中ESE3的表达量明显低于正常胰腺组织,ESE3的低表达与患者总生存期(OS)和无复发生存期(DFS)缩短密切相关(均P<0.05)。TCGA数据库分析表明ESE3表达高低与肿瘤转移相关(P<0.05);在乏氧和低糖条件下胰腺癌细胞侵袭能力增强,ESE3的表达降低,HIF-1α与GRP78的表达增加(P<0.05)。结论在乏氧和低糖条件下,胰腺癌细胞中ESE3的表达降低,促进胰腺癌细胞的侵袭。
ObjectiveTo observe the changes of epithelial specific ETS transcription factor 3(ESE3) expression in PDAC cell line under hypoxia and glucose deprived conditions, and its effect on the invasive ability of PDAC cells.MethodsPancreatic cancer tissues and 16 adjacent normal pancreas tissues from 99 patients with PDAC who underwent radical surgery from January 2005 to October 2014 in Tianjin Medical University Cancer Hospital were collected.Immunohistochemistry was used to analyze the expression of ESE3 in tumor tissues and adjacent normal tissues, and explore the relationship between ESE3 expression level and survival prognosis in PDAC patients. The relationship between ESE3 mRNA expression level and clinic pathological features of 147 patients with PDAC was further validated in TCGA database.The PANC-1 overexpressing ESE3 stably transfected cell line was constructed and validated. Transwell assay was used to observe the invasive ability of PDAC cells with overexpressing ESE3. The expressions of HIF-1α, GRP78 and ESE3 under hypoxic and glucose deprived conditions were detected by Western blot assay.ResultsThe expression of ESE3 was significantly lower in PDAC tissues than that in normal pancreatic tissues. The low expression of ESE3 was closely related to the shorten of overall survival(OS) and non-recurrence survival(DFS, P<0.05). The TCGA database demonstrated that ESE3 expression was associated with metastasis(P<0.05). The invasion ability of PDAC cells was increased under hypoxic and glucose deprived conditions, the expression of ESE3 was decreased, and the expressions of HIF-1α and GRP78 were increased(P<0.05).ConclusionUnder hypoxic and glucose deprived conditions, the expression of ESE3 is decreased in PDAC cells, which promotes PDAC cells invasion.
ObjectiveTo observe the changes of epithelial specific ETS transcription factor 3(ESE3) expression in PDAC cell line under hypoxia and glucose deprived conditions, and its effect on the invasive ability of PDAC cells.MethodsPancreatic cancer tissues and 16 adjacent normal pancreas tissues from 99 patients with PDAC who underwent radical surgery from January 2005 to October 2014 in Tianjin Medical University Cancer Hospital were collected.Immunohistochemistry was used to analyze the expression of ESE3 in tumor tissues and adjacent normal tissues, and explore the relationship between ESE3 expression level and survival prognosis in PDAC patients. The relationship between ESE3 mRNA expression level and clinic pathological features of 147 patients with PDAC was further validated in TCGA database.The PANC-1 overexpressing ESE3 stably transfected cell line was constructed and validated. Transwell assay was used to observe the invasive ability of PDAC cells with overexpressing ESE3. The expressions of HIF-1α, GRP78 and ESE3 under hypoxic and glucose deprived conditions were detected by Western blot assay.ResultsThe expression of ESE3 was significantly lower in PDAC tissues than that in normal pancreatic tissues. The low expression of ESE3 was closely related to the shorten of overall survival(OS) and non-recurrence survival(DFS, P<0.05). The TCGA database demonstrated that ESE3 expression was associated with metastasis(P<0.05). The invasion ability of PDAC cells was increased under hypoxic and glucose deprived conditions, the expression of ESE3 was decreased, and the expressions of HIF-1α and GRP78 were increased(P<0.05).ConclusionUnder hypoxic and glucose deprived conditions, the expression of ESE3 is decreased in PDAC cells, which promotes PDAC cells invasion.
引文
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[13]Takeuchi S,Baghdadi M,Tsuchikawa T,et al.Chemotherapyderived inflammatory responses accelerate the formation of immunosuppressive myeloid cells in the tissue microenvironment of human Pancreatic cancer[J].Cancer Res,2015,75(13):2629-2640.doi:10.1158/0008-5472.CAN-14-2921.
[14]Zhao T,Jiang W,Wang X,et al.ESE3 inhibits pancreatic cancer metastasis by upregulating E-cadherin[J].Cancer Res,2017,77(4):874-885.doi:10.1158/0008-5472.CAN-16-2170.
[2]Ko AH.Progress in the treatment of metastatic pancreatic cancer and the search for next opportunities[J].J Clin Oncol,2015,33(16):1779-1786.doi:10.1200/JCO.2014.59.7625.
[3]Cooper CD,Newman JA,Gileadi O.Recent advances in the structural molecular biology of Ets transcription factors:interactions,interfaces and inhibition[J].Biochem Soc Trans,2014,42(1):130-138.doi:10.1042/BST20130227.
[4]Lv Y,Sui F,Ma J,et al.Increased expression of EHF contributes to thyroid tumorigenesis through transcriptionally regulating HER2and HER3[J].Oncotarget,2016,7(36):57978-57990.doi:10.18632/oncotarget.11154.
[5]Pothuraju R,Rachagani S,Junker WM,et al.Pancreatic cancer associated with obesity and diabetes:An alternative approach for its targeting[J].J Exp Clin Cancer Res,2018,37(1):267-272.doi:10.1186/s13046-018-0963-4.
[6]Gao J,Aksoy BA,Dogrusoz U,et al.Integrative analysis of complex cancer genomics and clinical profiles using the c BioPortal[J].Sci Signal,2013,6(269):pl1.doi:10.1126/scisignal.2004088.
[7]Hwang HK,Jung MJ,Lee SH,et al.Adverse oncologic effects of intraoperative transfusion during pancreatectomy for left-sided pancreatic cancer:the need for strict transfusion policy[J].JHepatobiliary Pancreat Sci,2016,23(8):497-507.doi:10.1002/jhbp.368.
[8]Wang L,Xing J,Cheng R,et al.Abnormal localization and tumor suppressor function of epithelial tissue-specific transcription factor ESE3 in esophageal squamous cell carcinoma[J].PLoS One,2015,10(5):e0126319.doi:10.1371/journal.pone.0126319.
[9]Dallavalle C,Albino D,Civenni G,et al.MicroRNA-424 impairs ubiquitination to activate STAT3 and promote prostate tumor progression[J].J Clin Invest,2016,126(12):4585-4602.doi:10.1172/JCI86505.
[10]Albino D,Civenni G,Dallavalle C,et al.Activation of the Lin28/let-7 axis by loss of ESE3/EHF promotes a tumorigenic and stemlike phenotype in prostate cancer[J].Cancer Res,2016,76(12):3629-3643.doi:10.1158/0008-5472.CAN-15-2665.
[11]Zhu G,Zhou L,Liu H,et al.MicroRNA-224 promotes pancreatic cancer cell proliferation and migration by targeting the TXNIP-mediated HIF1αpathway[J].Cell Physiol and Biochem,2018,48(4):1735-1746.doi:10.1159/000492309.
[12]Zhao T,Ren H,Jia L,et al.Inhibition of HIF-1αby PX-478enhances the anti-tumor effect of gemcitabine by inducing immunogenic cell death in pancreatic ductal adenocarcinoma[J].Oncotarget,2015,6(4):2250-2262.doi:10.18632/oncotarget.2948.
[13]Takeuchi S,Baghdadi M,Tsuchikawa T,et al.Chemotherapyderived inflammatory responses accelerate the formation of immunosuppressive myeloid cells in the tissue microenvironment of human Pancreatic cancer[J].Cancer Res,2015,75(13):2629-2640.doi:10.1158/0008-5472.CAN-14-2921.
[14]Zhao T,Jiang W,Wang X,et al.ESE3 inhibits pancreatic cancer metastasis by upregulating E-cadherin[J].Cancer Res,2017,77(4):874-885.doi:10.1158/0008-5472.CAN-16-2170.