沈和德,杨文秀,褚明亮,易韦,张著学
SHEN Hede,YANG Wenxiu,CHU Mingliang,YI Wei,ZHANG Zhuxue

• 1:贵州医科大学基础医学院
• 2:贵州医科大学附属医院病理科
• 3:贵州医科大学附属人民医院病理科

摘要(Abstract)：

目的:探讨沉默高表达尾核同源盒转录因子2(CDX2)胃癌细胞中后对肝细胞核因子4α(HNF4α)表达水平的变化。方法:采用Western blot检测CDX2和HNF4α两种蛋白在AGS、BGC-823、MGC-803、MKN-45和SGC-7901五种胃癌细胞的表达,筛选出2者在同一种细胞蛋白中均高表达的胃癌细胞;选择高表达CDX2和HNF4α的AGS细胞,采用Western lot检测沉默CDX2表达后细胞中HNF4α蛋白表达情况。结果:Western blot结果显示,CDX2与HNF4a蛋白在AGS胃癌细胞均有较高表达;与阴性对照相比,经沉默CDX2表达后的AGS胃癌细胞中CDX2蛋白为低表达,HNF4α蛋白也为低表达,差异有统计学意义(P <0.05)。结论:在高表达CDX2的AGS胃癌细胞中沉默CDX2后,CDX2与HNF4α蛋白均呈低表达水平。
Objective: To investigate the expression level changes of HNF4α after silenced high expression CDX2 of gastric cancer cells. Methods: Western blot was adopted to detect the expression of CDX2 and HNF4α proteins in five gastric cancer cells( AGS,BGC-823,MGC-803,MKN-45,and SGC-7901) and screen the gastric cancer cells with high expression of both CDX2 and HNF4α in the same one. Selecting high expression CDX2 and HNF4α AGS cells,then Western blot was used to detect HNF4α protein expression after silenced expression of CDX2. Results: Western blot showed that CDX2 and HNF4 a proteins are highly expressed in AGS gastric cancer cells. Compared with the negative control,the expression of CDX2 in AGS gastric cancer cells was low after CDX2 gene silencing,and the expression of HNF4α protein was also low,differences were statistically significant( P < 0. 05). Conclusion: After silencing CDX2 in high expression CDX2 of AGS gastric cancer cell,CDX2 and HNF4α protein show low expression level.

关键词(KeyWords)： 胃肿瘤;基因沉默;尾核同源盒转录因子2蛋白;肝细胞核因子4α蛋白;蛋白免疫印迹
stomach neoplasms;gene silencing;CDX2 protein;HNF4α protein;western blot

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金地区基金项目(81560088);; 贵州省科技厅科研项目[黔科合J字(2015)2086];贵州省科技厅基金[LH字(2015)7108]

作者(Author): 沈和德,杨文秀,褚明亮,易韦,张著学
SHEN Hede,YANG Wenxiu,CHU Mingliang,YI Wei,ZHANG Zhuxue

DOI: 10.19367/j.cnki.2096-8388.2020.08.006

参考文献(References)：

• [1]JEMAL A,BRAY F,CENTER M M,et al.Global cancer statistics[J].Ca A Cancer Journal for Clinicians,2015,61(2):69-90．
• [2]LI Z X,KAMINISHI M.A comparison of gastric cancer between Japan and China[J].Gastric Cancer,2009,12(1):52-53．
• [3]MANDAL P K,CHAKRABARTI S,RAY A,et al.Mucin histochemistry of stomach in metaplasia and adenocarcinoma:an observation[J].Indian Journal of Medical&Paediatric Oncology Official Journal of Indian Society of Medical&Paediatric Oncology,2014,34 (4):229-233．
• [4]FILIPE M I,MUNOZ N,MATKO I,et al.Intestinal metaplasia types and the risk of gastric cancer:a cohort study in Slovenia[J].International Journal of Cancer,1994,57(3):324-329．
• [5]ROKKAS T,FILIPE M I,SLADEN G E.Detection of an increased incidence of early gastric cancer in patients with intestinal metaplasia type III who are closely followed up[J].Gut,1991,32(10):1110-1113．
• [6]PANTERIS V,NIKOLOPOULOU S,LOUNTOU A,et al.Diagnostic capabilities of high-definition white light endoscopy for the diagnosis of gastric intestinal metaplasia and correlation with histologic and clinical data[J].European Journal of Gastroenterology&Hepatology,2014,26(6):594-601．
• [7]CORREA P,HAENSZEL W,CUELLO C,et al.A model for gastric cancer epidemiology[J].Lancet,1975,2(7924):58．
• [8]GUZMAN P,ARAYA J,VILLASECA M,et al.Immunohistochemical expression of the E-cadherin-catenin complex in gastric cancer[J].Revista Médica De Chile,2006,134(8):1002-1009．
• [9]NAKAJIMA T,MAEKITA T,ODA I,et al.Higher methylation levels in gastric mucosae significantly correlate with higher risk of gastric cancers[J].Cancer Epidemiology,Biomarkers&Prevention:A Publication of The American Association for Cancer Research,Cosponsored by The American Society of Preventive Oncology,2006,15(11):2317．
• [10]方义湖，来茂德．结直肠肿瘤的新标记---CDX2[J]．国际遗传学杂志，2006,29(4):311-316．
• [11]MESQUITA P,JONCKHEERE N,ALMEIDA R,et al.Human MUC2 mucin gene is transcriptionally regulated by Cdx homeodomain proteins in gastrointestinal carcinoma cell lines.[J].Journal of Biological Chemistry,2003,278(51):51549．
• [12]SIBERG D G,SULLIVAN J,KANG E,et al.Cdx2 ectopic expression induces gastric intestinal metaplasia in transgenic mice[J].Gastroenterology,2002,122(3):689．
• [13]MUTOH H,HAKAMATA Y,SATO K,et al.Conversion of gastric mucosa to intestinal metaplasia in Cdx2-expressing transgenic mice[J].Biochem Biophys Res Commun,2002,294(2):470-479．
• [14]CHEN H X,CHEN M H,SONG X.Expression and significance of caudal type transcription factor-2 in gastric mucosa at different stages of gastric carcinogenesis[J].National Medical Journal of China,2006,86(4):247．
• [15]VANDER P R,BULT P,VOGELAAR I P,et al.HNF4A immunohistochemistry facilitates distinction between primary and metastatic breast and gastric carcinoma[J].Virchows Arch,2014,464:673-679．
• [16]SLADEK F M,ZHONG W M,LAI E,et al.Liver-enriched transcription factor HNF-4 is a novel member of the steroid hormone receptor superfamily[J].Genes&Development,1990,4(12):2353．
• [17]SLADEK F M.What are nuclear receptor ligands[J].Molecular&Cellular Endocrinology,2011,334(1-2):3-13．
• [18]KIM J,HOFFMAN J P,ALPAUGH R K,et al.An i P-SC line from human pancreatic ductal adenocarcinoma undergoes early to invasive stages of pancreatic cancer progression[J].Cell Rep,2013,3(6):2088-2099．
• [19]CHELLAPPA K,ROBERTSON G R,SLADEK F M.HNF4alpha:a new biomarker in colon cancer[J].Bromark med,2004,6:297-300．
• [20]SUGANO M,NAGASAKA T,SASAKI E,et al.HNF4a as a marker for invasive mucinous adenocarcinoma of the lung[J].Am J Surg Pathol,2013,37:211-218．
• [21]KOJIMA K,KISHIMOTO T,NAGAI Y,et al.The expression of hepatocyte nuclear factor-4alpha,a developmental regulator of visceral endoderm,correlates with the intestinal phenotype of gastric adenocarcinomas[J].Pathology,2006,38(6):548．
• [22]SAANDI T,BARAILLE,FDERBAL-WOLFROM L,et al.Regulation of the tumor suppressor homeogene Cdx2 by HNF4 in intestinal cancer[J].Oncogene,2013,32:3782-3788．
• [23]TARAVIRAS S,MONAGHAN A P,SCHUTZ G,et al.Characterization of the mouse HNF-4 gene and its expression during mouse embryogenesis[J].Mechanisms of Development,1994,48(2):67．
• [24]CHU M,CHANG Y,GUO Y,et al.Regulation and methylation of tumor suppressor miR-124 by androgen receptor in prostate cancer cells[J].Plos One,2015,10(4):e0116197．
• [25]FILIPE M I,MUNOZ N,MATKO I,et al.Intestinal metaplasia types and the risk of gastric cancer:a cohort study in Slovenia[J].International Journal of Cancer,1994,57(3):324．
• [26]GONZALEZ C A,SANZ-ANQUELA J M,GISBERT JP,et al.Utility of subtyping intestinal metaplasia as marker of gastric cancer risk.A review of the evidence[J].International Journal of Cancer,2013,133 (5):1023-1032．
• [27]YOU W C,LI J Y,BLOT W J,et al.Evolution of precancerous lesions in a rural Chinese population at high risk of gastric cancer[J].International Journal of Cancer,1999,83(5):615-619.

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