FOXA2在胰腺癌组织中的表达及其对胰腺癌细胞增殖和侵袭的影响
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摘要
目的探讨FOXA2在胰腺癌组织中的表达,以及特异性沉默FOXA2基因表达对PANC-1细胞增殖、迁移和侵袭能力的影响。方法选取2010年1月至2013年1月在郑州市中心医院择期行手术切除的胰腺癌患者71例,所有患者从手术时间开始进行随访,截止时间为2018年1月31日或患者死亡时间,采用免疫组化SP法检测胰腺癌和癌旁组织中FOXA2蛋白表达,生存分析采用Kaplan-Meier法和Log-Rank检验,影响患者预后的危险因素分析采用Cox比例风险回归模型。培养人胰腺癌PANC-1细胞,分为siRNAFOXA2组、siRNA-NC组和对照组,实时荧光定量PCR技术和Westernblotting法检测细胞中FOXA2mRNA和蛋白表达,CCK-8法检测细胞增殖活性,Transwell法检测细胞迁移和侵袭能力。结果胰腺癌组织中FOXA2蛋白阳性表达率(32.39%)低于癌旁组织(83.10%,P<0.001);胰腺癌组织中FOXA2蛋白阳性表达与TNM分期、淋巴结转移及脉管癌栓有关(P<0.05);Kaplan-Meier生存分析结果显示,阳性组患者平均生存时间高于阴性组,Log-Rank检验差异有统计学意义(χ2=11.135,P=0.001);Cox比例风险回归模型结果显示,淋巴结转移、脉管癌栓和FOXA2蛋白阴性表达是影响患者预后的风险因素(P<0.05);siRNAFOXA2组细胞中FOXA2 mRNA和蛋白表达量低于siRNA-NC组和对照组(P<0.05);siRNA-FOXA2组24 h、48 h、72 h和96 h时OD值均高于siRNA-NC组和对照组(P<0.05);siRNA-FOXA2组PANC-1和AsPC-1细胞中迁移细胞数和侵袭细胞数均高于siRNA-NC组和对照组(P<0.05)。结论胰腺癌组织中FOXA2蛋白呈低表达,是影响患者预后的风险因素;沉默FOXA2后会促进胰腺癌细胞增殖、迁移和侵袭。
Objective To investigate the expression of FOXA2 in pancreatic carcinoma, and to explore the effects of specific silencing of FOXA2 gene expression on the proliferation, migration and invasion of PANC-1 cells. Methods A total of 71 cases of patients with pancreatic carcinoma underwent elective surgical resection in Zhengzhou Central Hospital were selected from Jan. 2010 to Jan. 2013. All patients were followed-up from the time of surgery treatment, and the deadline was Jan. 31, 2018 or the time of death. Immunohistochemical SP method was used to detect the expressions of FOXA2 proteins in pancreatic carcinoma and adjacent tissues.Kaplan-Meier method and Log-Rank test was used for survival analysis. Cox proportional risk regression model was used to analysis the risk factors that affect patient prognosis. Human pancreatic carcinoma PANC-1 cells were cultured and divided into siRNA-FOXA2 group, siRNA-NC group and control group. Real-time fluorescence quantitative PCR and Western blotting were used to detect the expressions of FOXA2 mRNA and protein in cells. CCK-8 assay was used to detect cell proliferation activity. Transwell assay was used to detect cell migration and cell invasion. Results The positive expression rate of FOXA2 protein in pancreatic carcinoma tissues was 32.39%, which was lower than that in adjacent tissues(83.1%, P<0.001). The positive expression of FOXA2 protein in pancreatic carcinoma was related to TNM stage, lymph node metastasis and vascular tumor embolus(P<0.05). Kaplan-Meier survival analysis showed that the average survival time in the positive group was higher than that in the negative group, the Log-Rank test showed that the difference was statistically significant(X2=11.135, P=0.001). Cox proportional hazards regression model results showed that lymph node metastasis, vascular tumor embolus, and negative expression of FOXA2 protein were risk factors that affecting patient prognosis(P<0.05). The expression levels of FOXA2 mRNA and protein in the siRNA-FOXA2 group were lower than those in the siRNA-NC group and the control group(P<0.05). The OD values at 24 h, 48 h,72 h and 96 h in the siRNA-FOXA2 group were higher than those in the siRNA-NC group and the control group(P<0.05). The number of migrating cells and the number of invasive cells in the siRNA-FOXA2 group were higher than those in the siRNA-NC group and the control group(P<0.05). Conclusion The expression of FOXA2 protein in pancreatic carcinoma tissues is low, which is a risk factor influencing prognosis. Silencing of FOXA2 expression can promote proliferation, migration and invasion of pancreatic cancer cells.
Objective To investigate the expression of FOXA2 in pancreatic carcinoma, and to explore the effects of specific silencing of FOXA2 gene expression on the proliferation, migration and invasion of PANC-1 cells. Methods A total of 71 cases of patients with pancreatic carcinoma underwent elective surgical resection in Zhengzhou Central Hospital were selected from Jan. 2010 to Jan. 2013. All patients were followed-up from the time of surgery treatment, and the deadline was Jan. 31, 2018 or the time of death. Immunohistochemical SP method was used to detect the expressions of FOXA2 proteins in pancreatic carcinoma and adjacent tissues.Kaplan-Meier method and Log-Rank test was used for survival analysis. Cox proportional risk regression model was used to analysis the risk factors that affect patient prognosis. Human pancreatic carcinoma PANC-1 cells were cultured and divided into siRNA-FOXA2 group, siRNA-NC group and control group. Real-time fluorescence quantitative PCR and Western blotting were used to detect the expressions of FOXA2 mRNA and protein in cells. CCK-8 assay was used to detect cell proliferation activity. Transwell assay was used to detect cell migration and cell invasion. Results The positive expression rate of FOXA2 protein in pancreatic carcinoma tissues was 32.39%, which was lower than that in adjacent tissues(83.1%, P<0.001). The positive expression of FOXA2 protein in pancreatic carcinoma was related to TNM stage, lymph node metastasis and vascular tumor embolus(P<0.05). Kaplan-Meier survival analysis showed that the average survival time in the positive group was higher than that in the negative group, the Log-Rank test showed that the difference was statistically significant(X2=11.135, P=0.001). Cox proportional hazards regression model results showed that lymph node metastasis, vascular tumor embolus, and negative expression of FOXA2 protein were risk factors that affecting patient prognosis(P<0.05). The expression levels of FOXA2 mRNA and protein in the siRNA-FOXA2 group were lower than those in the siRNA-NC group and the control group(P<0.05). The OD values at 24 h, 48 h,72 h and 96 h in the siRNA-FOXA2 group were higher than those in the siRNA-NC group and the control group(P<0.05). The number of migrating cells and the number of invasive cells in the siRNA-FOXA2 group were higher than those in the siRNA-NC group and the control group(P<0.05). Conclusion The expression of FOXA2 protein in pancreatic carcinoma tissues is low, which is a risk factor influencing prognosis. Silencing of FOXA2 expression can promote proliferation, migration and invasion of pancreatic cancer cells.
引文
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[10]Le Gallo M,Rudd ML,Urick ME,et al.The FOXA2 transcription factor is frequently somatically mutated in uterine carcinosarcomas and carcinomas[J].Cancer,2018,124(1):65-73.
[11]李正平,张宇,罗道蕴,等.叉头框转录因子A2对肝癌细胞迁移和侵袭能力的影响[J].广西医学,2015,37(6):755-757.
[12]Ashaie MA,Chowdhury EH.Cadherins:the superfamily critically involved in breast cancer[J].Curr Pharm Des,2016,22(5):616-638.
[13]Li CM,Gocheva V,Oudin MJ,et al.Foxa2 and Cdx2 cooperate with Nkx2-1 to inhibit lung adenocarcinoma metastasis[J].Genes Dev,2015,29(17):1850-1862.
[14]彭俊文,陈健,许海林,等.FOXA2在人胃癌中的表达及预后意义[J].实用肿瘤杂志,2016,31(2):150-153.
[15]王曦,曹佳,王丹妮,等.过表达FOXA2对结直肠癌LOVO细胞增殖和凋亡的影响[J].宁夏医科大学学报,2018,40(4):378-383.
[16]Shi W,Wang X,Ruan L,et al.MiR-200a promotes epithelial-mesenchymal transition of endometrial cancer cells by negatively regulating FOXA2 expression[J].Pharmazie,2017,72(11):694-699.
[2]Zhang L,Sanagapalli S,Stoita A.Challenges in diagnosis of pancreatic cancer[J].World J Gastroenterol,2018,24(19):2047-2060.
[3]Benzel J,Fendrich V.Chemoprevention and treatment of pancreatic cancer:update and review of the literature[J].Digestion,2018,97(4):275-287.
[4]King J,Bouvet M,Singh G,et al.Improving theranostics in pancreatic cancer[J].J Surg Oncol,2017,116(1):104-113.
[5]姚磊,刘泰荣,陈华.FoxA家族与恶性肿瘤的发生发展关系研究进展[J].转化医学电子杂志,2018,5(2):45-48.
[6]Smith B,Neff R,Cohn DE,et al.The mutational spectrum of FOXA2 in endometrioid endometrial cancer points to a tumor suppressor role[J].Gynecol Oncol,2016,143(2):398-405.
[7]胡伟,王磊,孙磊,等.IQ结构域GTP酶激活蛋白1在胰腺癌中表达及与预后的相关性研究[J].中国现代医学杂志,2017,27(30):41-45.
[8]潘成文,朱励民,吴育连.胰腺癌肝转移外科治疗进展[J].中华普通外科杂志,2017,32(3):279-281.
[9]Han H,Wang D,Yang M,et al.High expression of RACK1is associated with poor prognosis in patients with pancreatic ductal adenocarcinoma[J].Oncol Lett,2018,15(2):2073-2078.
[10]Le Gallo M,Rudd ML,Urick ME,et al.The FOXA2 transcription factor is frequently somatically mutated in uterine carcinosarcomas and carcinomas[J].Cancer,2018,124(1):65-73.
[11]李正平,张宇,罗道蕴,等.叉头框转录因子A2对肝癌细胞迁移和侵袭能力的影响[J].广西医学,2015,37(6):755-757.
[12]Ashaie MA,Chowdhury EH.Cadherins:the superfamily critically involved in breast cancer[J].Curr Pharm Des,2016,22(5):616-638.
[13]Li CM,Gocheva V,Oudin MJ,et al.Foxa2 and Cdx2 cooperate with Nkx2-1 to inhibit lung adenocarcinoma metastasis[J].Genes Dev,2015,29(17):1850-1862.
[14]彭俊文,陈健,许海林,等.FOXA2在人胃癌中的表达及预后意义[J].实用肿瘤杂志,2016,31(2):150-153.
[15]王曦,曹佳,王丹妮,等.过表达FOXA2对结直肠癌LOVO细胞增殖和凋亡的影响[J].宁夏医科大学学报,2018,40(4):378-383.
[16]Shi W,Wang X,Ruan L,et al.MiR-200a promotes epithelial-mesenchymal transition of endometrial cancer cells by negatively regulating FOXA2 expression[J].Pharmazie,2017,72(11):694-699.