远端上游元件结合蛋白1在幽门螺杆菌相关性胃炎中的表达及机制研究
|
摘要
目的研究远端上游元件结合蛋白1(far upstream element-binding protein 1,FUBP1)在幽门螺杆菌(Helicobacter pylori,H. pylori)相关性胃炎中的表达,并初步探讨H. pylori引起FUBP1表达变化的可能机制。方法分别采用实时荧光定量逆转录聚合酶链式反应(RT-q PCR)、蛋白质印迹法(Western blotting)和免疫组织化学技术检测H. pylori阴性和H. pylori阳性胃炎组织中FUBP1mRNA和蛋白的表达。国际标准测序株H. pylori 26695及其提取的LPS分别刺激AGS、SGC7901细胞,RT-q PCR和Western blotting检测FUBP1 mRNA和蛋白的表达。分别使用H. pylori和H. pylori-LPS灌胃,构建小鼠感染模型,于灌胃结束后4周、8周取小鼠胃黏膜组织,RT-q PCR和Western blotting检测FUBP1 mRNA和蛋白表达。结果 H. pylori阳性组织中FUBP1 mRNA及蛋白表达水平均低于H. pylori阴性组织,FUBP1蛋白水平的表达差异有统计学意义(P <0. 05)。H. pylori刺激AGS和SGC7901细胞后FUBP1mRNA和蛋白水平降低,且呈时间依赖性,H. pylori-LPS刺激细胞后,FUBP1 mRNA和蛋白水平表达降低,但无时间及浓度依赖性。动物模型中,与对照组相比,H. pylori组和H. pylori-LPS组的FUBP1 mRNA和蛋白表达水平均明显降低,差异有统计学意义(P <0. 05)。结论 FUBP1在H. pylori相关性胃炎中表达下降,H. pylori和H. pylori毒力因子LPS均能诱导FUBP1下调,FUBP1可能成为评价慢性胃炎患者H. pylori感染的一个潜在指标。
Objective To investigate the expression of far upstrearn element-binding protein 1(FUBP1) in H. pyloriassociated gastritis and to explore the possible mechanism of H. pylori inducing FUBP1 expression. Methods The expressions of FUBP1 mRNA and protein in H. pylori negative and positive gastritis tissues were detected by RT-q PCR,Western blotting and immunohistochemistry,respectively. The expressions of FUBP1 mRNA and protein were detected in human gastric epithelial cancer cells line AGS and SGC7901 infected by standard H. pylori 26695 and its extracted Lipopolysaccharide. The infected animal models were established by intragastric administration of H. pylori and H. pyloriLPS,respectively. The gastric mucosal tissues of mice were taken in the 4 th and 8 th weeks,and the expressions of FUBP1 mRNA and protein were detected by RT-q PCR and Western blotting. Results The expressions of FUBP1 mRNA and protein in H. pylori positive tissues were down-regulated compared with H. pylori negative tissues. The difference of FUBP1 protein expression was statistically significant(P < 0. 05). In H. pylori-stimulated AGS and SGC7901 cells,the expressions of FUBP1 mRNA and protein were gradually decreased and had time dependent. In AGS and SGC7901 cells stimulated with H. pylori-LPS,the expressions of FUBP1 mRNA and protein were decreased,but there was no time and concentration-dependent change in FUBP1. Compared with control group,the expressions of FUBP1 mRNA and proteins in H. pylori group and H. pylori-LPS group were significantly decreased in the 4 th and 8 th weeks infected animal models.The difference was statistically significant(P < 0. 05). Conclusion The expression of FUBP1 in H. pylori-associated gastritis decreases. H. pylori and H. pylori virulence factor LPS can induce down regulation of FUBP1. FUBP1 may be a potential indicator of H. pylori infection in patients with chronic gastritis.
Objective To investigate the expression of far upstrearn element-binding protein 1(FUBP1) in H. pyloriassociated gastritis and to explore the possible mechanism of H. pylori inducing FUBP1 expression. Methods The expressions of FUBP1 mRNA and protein in H. pylori negative and positive gastritis tissues were detected by RT-q PCR,Western blotting and immunohistochemistry,respectively. The expressions of FUBP1 mRNA and protein were detected in human gastric epithelial cancer cells line AGS and SGC7901 infected by standard H. pylori 26695 and its extracted Lipopolysaccharide. The infected animal models were established by intragastric administration of H. pylori and H. pyloriLPS,respectively. The gastric mucosal tissues of mice were taken in the 4 th and 8 th weeks,and the expressions of FUBP1 mRNA and protein were detected by RT-q PCR and Western blotting. Results The expressions of FUBP1 mRNA and protein in H. pylori positive tissues were down-regulated compared with H. pylori negative tissues. The difference of FUBP1 protein expression was statistically significant(P < 0. 05). In H. pylori-stimulated AGS and SGC7901 cells,the expressions of FUBP1 mRNA and protein were gradually decreased and had time dependent. In AGS and SGC7901 cells stimulated with H. pylori-LPS,the expressions of FUBP1 mRNA and protein were decreased,but there was no time and concentration-dependent change in FUBP1. Compared with control group,the expressions of FUBP1 mRNA and proteins in H. pylori group and H. pylori-LPS group were significantly decreased in the 4 th and 8 th weeks infected animal models.The difference was statistically significant(P < 0. 05). Conclusion The expression of FUBP1 in H. pylori-associated gastritis decreases. H. pylori and H. pylori virulence factor LPS can induce down regulation of FUBP1. FUBP1 may be a potential indicator of H. pylori infection in patients with chronic gastritis.
引文
[1] SELGRAD M,BORNSCHEIN J,ROKKAS T,et al. Helicobacter pylori:gastric cancer and extragastric intestinal malignancies[J]. Helicobacter,2012,17 Suppl 1:30-35. DOI:10. 1111/j. 1523-5378.2012. 00980. x.
[2] WECK M N,GAO L,BRENNER H. Helicobacter pylori infection and chronic atrophic gastritis:associations according to severity of disease[J]. Epidemiology,2009,20(4):569-574. DOI:10. 1097/EDE. 0b013e3181a3d5f4.
[3] BENJAMIN L R,CHUNG H J,SANFORD S,et al. Hierarchical mechanisms build the DNA-binding specificity of FUSE binding protein[J].Proc Natl Acad Sci U S A,2008,105(47):18296-18301. DOI:10.1073/pnas. 0803279105.
[4] ZHANG J,CHEN Q M. Far upstream element binding protein 1:a commander of transcription,translation and beyond[J]. Oncogene,2013,32(24):2907-2916. DOI:10. 1038/onc. 2012. 350.
[5] ZHANG F,TIAN Q,WANG Y. Far upstream element-binding protein1(FUBP1)is overexpressed in human gastric cancer tissue compared to non-cancerous tissue[J]. Onkologie,2013,36(11):650-655.DOI:10. 1159/000355659.
[6]李丹,黄宝俊,赵丹懿.远端上游元件结合蛋白1在胃癌中的表达及其对胃癌细胞增殖和侵袭能力的影响[J].中国医科大学学报,2017,46(7):587-590. DOI:10. 12007/j. issn. 0258-4646.2017. 07. 003.LI D,HUANG B J,ZHAO D Y. Expression of far upstream element binding protein 1 in gastric cancer and its effect on proliferation and invasion of gastric cancer cells[J]. Journal of China Medical University,2017,46(7):587-590. DOI:10. 12007/j. issn. 0258-4646. 2017.07. 003.
[7] YE F,TANG C,SHI W,et al. A MDM2-dependent positive-feedback loop is involved in inhibition of miR-375 and miR-106b induced by Helicobacter pylori lipopolysaccharide[J]. Int J Cancer,2015,136(9):2120-2131. DOI:10. 1002/ijc. 29268.
[8]徐良中,杨文涛.免疫组织化学反应结果的判断标准[J].中国癌症杂志,1996,6(4):229-231.
[9] DIACONU S,PREDESCU A,MOLDOVEANU A,et al. Helicobacter pylori infection:old and new[J]. J Med Life,2017,10(2):112-117.
[10]高鹏,王玉平,周永宁.胃癌与幽门螺杆菌相关性研究进展[J].胃肠病学和肝病学杂志,2016,25(1):1-4. DOI:10. 3969/j.issn. 1006-5709. 2016. 01. 001.GAO P,WANG Y P,ZHOU Y N. Research progress of the correlation between gastric cancer and Helicobacter pylori[J]. Chin J Gastroenterol Hepatol,2016,25(1):1-4. DOI:10. 3969/j. issn. 1006-5709. 2016. 01. 001.
[11]刘文忠,谢勇,陆红,等.第五次全国幽门螺杆菌感染处理共识报告[J].胃肠病学,2017,37(6):346-360. DOI:10. 3960/cma. j. issn. 0254-1432. 2017. 06. 002.
[12] LUO Y H,YAN J,MAO Y F. Helicobacter pylori lipopolysaccharide:biological activities in vitro and in vivo,pathological correlation to human chronic gastritis and peptic ulcer[J]. World J Gastroenterol,2004,10(14):2055-2059.
[13] MATTSBY-BALTZER I,MIELNICZUK Z,LARSSON L,et al. Lipid A in Helicobacter pylori[J]. Infect Immun,1992,60(10):4383-4387.
[14] MORAN A P. Helicobacter pylori lipopolysaccharide-mediated gastric and extragastric pathology[J]. J Physiol Pharmacol,1999,50(5):787-805.
[15] JIA M Y,WANG Y J. Far upstream element-binding protein 1(FUBP1)expression differs between human colorectal cancer and non-cancerous tissue[J]. Neoplasma,2014,61(5):533-540.DOI:10. 4149/neo_2014_065.
[16] TANG Q,XIA W,JI Q,et al. Role of far upstream element binding protein 1 in colonic epithelial disruption during dextran sulphate sodium-induced murine colitis[J]. Int J Clin Exp Pathol,2014,7(5):2019-2031.
[2] WECK M N,GAO L,BRENNER H. Helicobacter pylori infection and chronic atrophic gastritis:associations according to severity of disease[J]. Epidemiology,2009,20(4):569-574. DOI:10. 1097/EDE. 0b013e3181a3d5f4.
[3] BENJAMIN L R,CHUNG H J,SANFORD S,et al. Hierarchical mechanisms build the DNA-binding specificity of FUSE binding protein[J].Proc Natl Acad Sci U S A,2008,105(47):18296-18301. DOI:10.1073/pnas. 0803279105.
[4] ZHANG J,CHEN Q M. Far upstream element binding protein 1:a commander of transcription,translation and beyond[J]. Oncogene,2013,32(24):2907-2916. DOI:10. 1038/onc. 2012. 350.
[5] ZHANG F,TIAN Q,WANG Y. Far upstream element-binding protein1(FUBP1)is overexpressed in human gastric cancer tissue compared to non-cancerous tissue[J]. Onkologie,2013,36(11):650-655.DOI:10. 1159/000355659.
[6]李丹,黄宝俊,赵丹懿.远端上游元件结合蛋白1在胃癌中的表达及其对胃癌细胞增殖和侵袭能力的影响[J].中国医科大学学报,2017,46(7):587-590. DOI:10. 12007/j. issn. 0258-4646.2017. 07. 003.LI D,HUANG B J,ZHAO D Y. Expression of far upstream element binding protein 1 in gastric cancer and its effect on proliferation and invasion of gastric cancer cells[J]. Journal of China Medical University,2017,46(7):587-590. DOI:10. 12007/j. issn. 0258-4646. 2017.07. 003.
[7] YE F,TANG C,SHI W,et al. A MDM2-dependent positive-feedback loop is involved in inhibition of miR-375 and miR-106b induced by Helicobacter pylori lipopolysaccharide[J]. Int J Cancer,2015,136(9):2120-2131. DOI:10. 1002/ijc. 29268.
[8]徐良中,杨文涛.免疫组织化学反应结果的判断标准[J].中国癌症杂志,1996,6(4):229-231.
[9] DIACONU S,PREDESCU A,MOLDOVEANU A,et al. Helicobacter pylori infection:old and new[J]. J Med Life,2017,10(2):112-117.
[10]高鹏,王玉平,周永宁.胃癌与幽门螺杆菌相关性研究进展[J].胃肠病学和肝病学杂志,2016,25(1):1-4. DOI:10. 3969/j.issn. 1006-5709. 2016. 01. 001.GAO P,WANG Y P,ZHOU Y N. Research progress of the correlation between gastric cancer and Helicobacter pylori[J]. Chin J Gastroenterol Hepatol,2016,25(1):1-4. DOI:10. 3969/j. issn. 1006-5709. 2016. 01. 001.
[11]刘文忠,谢勇,陆红,等.第五次全国幽门螺杆菌感染处理共识报告[J].胃肠病学,2017,37(6):346-360. DOI:10. 3960/cma. j. issn. 0254-1432. 2017. 06. 002.
[12] LUO Y H,YAN J,MAO Y F. Helicobacter pylori lipopolysaccharide:biological activities in vitro and in vivo,pathological correlation to human chronic gastritis and peptic ulcer[J]. World J Gastroenterol,2004,10(14):2055-2059.
[13] MATTSBY-BALTZER I,MIELNICZUK Z,LARSSON L,et al. Lipid A in Helicobacter pylori[J]. Infect Immun,1992,60(10):4383-4387.
[14] MORAN A P. Helicobacter pylori lipopolysaccharide-mediated gastric and extragastric pathology[J]. J Physiol Pharmacol,1999,50(5):787-805.
[15] JIA M Y,WANG Y J. Far upstream element-binding protein 1(FUBP1)expression differs between human colorectal cancer and non-cancerous tissue[J]. Neoplasma,2014,61(5):533-540.DOI:10. 4149/neo_2014_065.
[16] TANG Q,XIA W,JI Q,et al. Role of far upstream element binding protein 1 in colonic epithelial disruption during dextran sulphate sodium-induced murine colitis[J]. Int J Clin Exp Pathol,2014,7(5):2019-2031.