IBP与p53蛋白在胃腺癌中的表达及临床意义
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摘要
目的探讨干扰素调节因子4结合蛋白(IRF-4 binding protein,IBP)在胃腺癌细胞及临床病理组织中的表达、与p53蛋白表达的相关性及临床意义。方法通过免疫荧光、Real-time PCR和Western blot检测胃腺癌细胞株MKN-45、SGC-7901中IBP的表达情况;收集2015-2016年在本科确诊的胃腺癌HE切片对应存档蜡块150例,采用免疫组化EnVision法检测并标记IBP在150例胃腺癌标本中的表达,并检测突变型p53蛋白的表达,分析IBP在胃腺癌组织中的表达及其与胃腺癌各临床病理资料因子相应的关系,采用Spearman等级相关分析IBP与p53蛋白表达的相关性。结果免疫荧光检测显示IBP主要表达在胃腺癌细胞株MKN-45的胞质和胞膜中,主要以胞膜表达为主。Western blot及Real-time PCR均证实胃腺癌细胞株MKN-45、SGC-7901中均有IBP的表达,且在MKN-45细胞中的表达高于SGC-7901细胞(P <0. 05)。在胃腺癌组织中,IBP的阳性表达率显著高于正常胃黏膜组织(P <0. 01);p53蛋白的阳性表达率也显著高于正常胃黏膜组织(P <0. 01); IBP阳性表达与病理分级呈正相关(P <0. 05),与患者年龄、性别、胃壁浸润深度、TNM分期、Lauren分型及肿瘤部位无相关性(P> 0. 05); p53蛋白的阳性表达与患者的年龄、性别、病理分级、Lauren分型具有相关性(P <0. 05),与患者胃壁浸润深度、TNM分期及肿瘤部位无相关性(P> 0. 05);在胃腺癌中IBP与p53蛋白表达没有显著的相关关系(r=0. 013,P> 0. 05)。结论 IBP在胃腺癌细胞株及临床病理组织中高表达,主要表达于胞膜、胞质;p53蛋白在胃腺癌临床病理组织中高表达,主要表达于胞核; IBP在胃腺癌组织中高表达与患者病理分级正相关,在胃腺癌组织中IBP与p53蛋白没有明显的相关关系。
Objective To investigate the expression of IRF-4 binding protein( IBP) in gastric adenocarcinoma,and explore its correlation with p53 expression and clinical significance. Methods The expression of IBP in gastric adenocarcinoma cell lines MKN-45 and SGC-7901 was detected with immunofluorescence assay,real-time PCR and Western blotting. Immunohistochemical staining with the aid of EnVision system was used to detect and mark the expression of IBP in 150 cases of gastric adenocarcinoma identified in our department from 2015 to 2016. The expression of mutant p53 protein was also detected. The expression of IBP in gastric adenocarcinoma and its relationship with the clinical pathological data were analyzed,and the correlation between IBP and p53 expression was tested by Spearman grade correlation analysis. Results Immunofluorescence assay showed that IBP was expressed in the cytoplasm and membrane of the MKN-45 cells,mainly in the latter. Western blotting and real-time PCR confirmed that IBP was expressed in the MKN-45 and SGC-7901 cells,and the expression level was significantly higher in the former than the latter cells( P < 0. 05). The positive expression rate of IBP was obviously higher in the gastric adenocarcinoma tissues than the adjacent normal tissues( P < 0. 01),so was that of p53( P < 0. 01). The positive expression of IBP was positively correlated with pathological grade( P < 0. 05),but not with the patient's age,gender,gastric wall invasion depth,TNM stage,Lauren type or tumor site( P > 0. 05). The positive expression of p53 was correlated with age,sex,pathological grade,and Lauren type( P < 0. 05),but not with depth of gastric wall invasion,TNM stage or tumor site( P > 0. 05). No significant correlation was seen between the expression of IBP and p53 in gastric adenocarcinoma( r = 0. 013,P > 0. 05). Conclusion IBP is highly expressed in the gastric adenocarcinoma cell lines and gastric adenocarcinoma tissues,mainly in the cytoplasm and membrane,and p53 protein was highly expressed in the tumor tissues,mainly in the nucleus. The high expression of IBP in gastric adenocarcinoma is positively correlated with pathological grade,and there was no significant correlation between their expression.
Objective To investigate the expression of IRF-4 binding protein( IBP) in gastric adenocarcinoma,and explore its correlation with p53 expression and clinical significance. Methods The expression of IBP in gastric adenocarcinoma cell lines MKN-45 and SGC-7901 was detected with immunofluorescence assay,real-time PCR and Western blotting. Immunohistochemical staining with the aid of EnVision system was used to detect and mark the expression of IBP in 150 cases of gastric adenocarcinoma identified in our department from 2015 to 2016. The expression of mutant p53 protein was also detected. The expression of IBP in gastric adenocarcinoma and its relationship with the clinical pathological data were analyzed,and the correlation between IBP and p53 expression was tested by Spearman grade correlation analysis. Results Immunofluorescence assay showed that IBP was expressed in the cytoplasm and membrane of the MKN-45 cells,mainly in the latter. Western blotting and real-time PCR confirmed that IBP was expressed in the MKN-45 and SGC-7901 cells,and the expression level was significantly higher in the former than the latter cells( P < 0. 05). The positive expression rate of IBP was obviously higher in the gastric adenocarcinoma tissues than the adjacent normal tissues( P < 0. 01),so was that of p53( P < 0. 01). The positive expression of IBP was positively correlated with pathological grade( P < 0. 05),but not with the patient's age,gender,gastric wall invasion depth,TNM stage,Lauren type or tumor site( P > 0. 05). The positive expression of p53 was correlated with age,sex,pathological grade,and Lauren type( P < 0. 05),but not with depth of gastric wall invasion,TNM stage or tumor site( P > 0. 05). No significant correlation was seen between the expression of IBP and p53 in gastric adenocarcinoma( r = 0. 013,P > 0. 05). Conclusion IBP is highly expressed in the gastric adenocarcinoma cell lines and gastric adenocarcinoma tissues,mainly in the cytoplasm and membrane,and p53 protein was highly expressed in the tumor tissues,mainly in the nucleus. The high expression of IBP in gastric adenocarcinoma is positively correlated with pathological grade,and there was no significant correlation between their expression.
引文
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[20]简从相,李晨军,黄晓山. IRF-4结合蛋白对人口腔鳞癌细胞生长作用的研究[J].西南国防医药,2012,22(5):470-472. DOI:10. 3969/j. issn. 1004-0188. 2012. 05. 002.JIAN C X,LI C J,HUANG X S. Effects of IRF-4 binding protein on cell growth of oral squamous cell carcinoma[J].Med J Nat Defending Forces Southwest China,2012,22(5):470-472. DOI:10. 3969/j. issn. 1004-0188. 2012.05. 002.
[21]熊剑,简从相,李鹏,等. IRF-4结合蛋白对人涎腺腺样囊性癌细胞增殖的抑制作用[J].第三军医大学学报,2011,33(6):554-557. DOI:10. 16016/j. 1000-5404.2011. 06. 006.XIONG J,JIAN C X,LI P,et al. Inhibitory effect of IRF-4binding protein on proliferation of human salivary adenoid cystic carcinoma cells[J]. J Third Mil Med Univ,2011,33(6):554-557. DOI:10. 16016/j. 1000-5404. 2011. 06.006.
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[23] WARAYA M,YAMASHITA K,EMA A,et al. Exclusive association of p53 mutation with super-high methylation of tumor suppressor genes in the p53 pathway in a unique gastric cancer phenotype[J]. PLo S ONE,2015,10(10):e0139902. DOI:10. 1371/journal. pone. 0139902.
[2] DURAES C,ALMEIDA G M,SERUCA R,et al. Biomarkers for gastric cancer:prognostic,predictive or targets of therapy?[J]. Virchows Arch,2014,464(3):367-378. DOI:10. 1007/s00428-013-1533-y.
[3] BANIAK N,SENGER J L,AHMED S,et al. Gastric biomarkers:a global review[J]. World J Surg Oncol,2016,14(1):212. DOI:10. 1186/s12957-016-0969-3.
[4] BECART S,CHARVET C,CANONIGO BALANCIO A J,et al.SLAT regulates Th1 and Th2 inflammatory responses by controlling Ca2+/NFAT signaling[J]. J Clin Invest,2007,117(8):2164-2175. DOI:10. 1172/JCI31640.
[5] FANZO J C,YANG W,JANG S Y,et al. Loss of IRF-4-binding protein leads to the spontaneous development of systemic autoimmunity[J]. J Clin Invest,2006,116(3):703-714. DOI:10. 1172/JCI24096.
[6] LI P,ZHANG Z,WANG Q,et al. The ectopic expression of IFN regulatory factor 4-binding protein is correlated with the malignant behavior of human breast cancer cells[J]. Int Immunopharmacol,2009,9(7/8):1002-1009. DOI:10.1016/j. intimp. 2009. 04. 008.
[7] ZHANG Z,WANG Q,LI P,et al. Overexpression of the Interferon regulatory factor 4-binding protein in human colorectal cancer and its clinical significance[J]. Cancer Epidemiol,2009,33(2):130-136. DOI:10. 1016/j. canep. 2009.05. 004.
[8] MULLER P A,VOUSDEN K H. p53 mutations in cancer[J].Nat Cell Biol,2013,15(1):2-8. DOI:10. 1038/ncb2641.
[9] FANG L,GAO H,ZHANG W,et al. Resveratrol alleviates nerve injury after cerebral ischemia and reperfusion in mice by inhibiting inflammation and apoptosis[J]. Int J Clin Exp Med,2015,8(3):3219-3226.
[10] PRASS K,SCHARFF A,RUSCHER K,et al. Hypoxiainduced stroke tolerance in the mouse is mediated by erythropoietin[J]. Stroke,2003,34(8):1981-1986. DOI:10.1161/01. STR. 0000080381. 76409. B2.
[11] YANG M,YUAN F,LI P,et al. Interferon regulatory factor4 binding protein is a novel p53 target gene and suppresses cisplatin-induced apoptosis of breast cancer cells[J]. Mol Cancer,2012,11:54. DOI:10. 1186/1476-4598-11-54.
[12]赵志华,田研,陈奎生.多发性骨髓瘤中c-myc和MDM2蛋白的表达及其临床意义[J].临床与实验病理学杂志,2017,33(2):165-168. DOI:10. 13315/j. cnki. cjcep.2017. 02. 011.ZHAO Z H,TIAN Y,CHEN K S. Expression of c-myc and MDM2 protein in multiple myeloma and their clinical significance[J]. Chin J Clin Exp Pathol,2017,33(2):165-168. DOI:10. 13315/j. cnki. cjcep. 2017. 02. 011.
[13] GUPTA S,LEE A,HU C,et al. Molecular cloning of IBP,a SWAP-70 homologous GEF,which is highly expressed in the immune system[J]. Hum Immunol,2003,64(4):389-401. DOI:10. 1016/S0198-8859(03)00024-7.
[14] VEGA F M,RIDLEY A J. Rho GTPases in cancer cell biology[J]. FEBS Lett,2008,582(14):2093-2101. DOI:10. 1016/j. febslet. 2008. 04. 039.
[15] MAVRAKIS K J,MCKINLAY K J,JONES P,et al.DEF6,a novel PH-DH-like domain protein,is an upstream activator of the Rho GTPases Rac1,Cdc42,and Rho A[J].Exp Cell Res,2004,294(2):335-344. DOI:10. 1016/j.yexcr. 2003. 12. 004.
[16] JAFFE A B,HALL A. Rho GTPases:Biochemistry and biology[J]. Annu Rev Cell Dev Biol,2005,21:247-269. DOI:10. 1146/annurev. cellbio. 21. 020604. 150721.
[17] BOS J L,REHMANN H,WITTINGHOFER A. GEFs and GAPs:Critical elements in the control of small G proteins[J]. Cell,2007,129(5):865-877. DOI:10. 1016/j.cell. 2007. 05. 018.
[18] CHARDIN P. Function and regulation of Rnd proteins[J].Nat Rev Mol Cell Biol,2006,7(1):54-62. DOI:10.1038/nrm1788.
[19] ZHANG Z,YANG M,CHEN R,et al. IBP regulates epithelial-to-mesenchymal transition and the motility of breast cancer cells via Rac1,Rho A and Cdc42 signaling pathways[J]. Oncogene,2014,33(26):3374-3382. DOI:10.1038/onc. 2013. 337.
[20]简从相,李晨军,黄晓山. IRF-4结合蛋白对人口腔鳞癌细胞生长作用的研究[J].西南国防医药,2012,22(5):470-472. DOI:10. 3969/j. issn. 1004-0188. 2012. 05. 002.JIAN C X,LI C J,HUANG X S. Effects of IRF-4 binding protein on cell growth of oral squamous cell carcinoma[J].Med J Nat Defending Forces Southwest China,2012,22(5):470-472. DOI:10. 3969/j. issn. 1004-0188. 2012.05. 002.
[21]熊剑,简从相,李鹏,等. IRF-4结合蛋白对人涎腺腺样囊性癌细胞增殖的抑制作用[J].第三军医大学学报,2011,33(6):554-557. DOI:10. 16016/j. 1000-5404.2011. 06. 006.XIONG J,JIAN C X,LI P,et al. Inhibitory effect of IRF-4binding protein on proliferation of human salivary adenoid cystic carcinoma cells[J]. J Third Mil Med Univ,2011,33(6):554-557. DOI:10. 16016/j. 1000-5404. 2011. 06.006.
[22] HOTFILDER M,BAXENDALE S,CROSS M A,et al. Def-2,-3,-6 and-8,novel mouse genes differentially expressed in the haemopoietic system[J]. Br J Haematol,1999,106:335-344. DOI:10. 1046/j. 1365-2141. 1999. 01551. x.
[23] WARAYA M,YAMASHITA K,EMA A,et al. Exclusive association of p53 mutation with super-high methylation of tumor suppressor genes in the p53 pathway in a unique gastric cancer phenotype[J]. PLo S ONE,2015,10(10):e0139902. DOI:10. 1371/journal. pone. 0139902.