肠型与弥漫型胃癌中Sonic hedgehog与Wnt3A/β-catenin的表达
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摘要
目的:观察音猬因子(Shh)、Wnt3A/β-catenin(β-连环蛋白)及分泌型卷曲相关蛋白-1(Sfrp1)在慢性浅表性胃炎(CSG)、肠型胃癌(IGC)及弥漫型胃癌(DGC)中表达特点,并研究它们的相关性,初步揭示这两种胃癌有完全不同的发病机制.方法:运用免疫组织化学法、Western Blotting方法检测30例CSG(作为对照组)、42例IGC及42例DGC标本中Shh、Wnt3A/β-catenin及Sfrp1蛋白表达特点,比较每种蛋白在各组中表达的差异,并分析它们之间的相关性.结果:免疫组织化学法检测发现:Shh主要表达在细胞浆和细胞膜.在CSG中Shh多阳性或强阳性表达;在IGC中,Shh大多表达减少,呈弱阳性表达,部分甚至不表达;在DGC中,Shh多呈强阳性表达.Shh在3组之间的表达有统计学差异(H=46.064,P<0.001).CSG中Shh表达水平明显高于IGC(P<0.001),显著低于DGC(P<0.05).Shh在DGC中表达水平显著高于IGC(P<0.001).在CSG中,β-catenin主要表达在细胞膜,细胞浆一般不染色或局部淡染,胞核不染色.在IGC中,β-catenin表达水平显著增高,并可观察到胞质与胞核中异位表达,但在DGC中却表达显著减少,有一些甚至不表达.β-catenin在3组之间的表达有统计学差异(H=38.629,P<0.001).β-catenin在CSG中表达水平显著低于IGC(P<0.05),显著高于DGC(P<0.001).β-catenin在IGC中表达显著高于DGC(P<0.001).蛋白免疫印迹试验检测发现:Shh在3组之间的表达有统计学差异(F=739.731,P<0.001).与CSG组比较,Shh蛋白在IGC组表达水平显著减少,在DGC组表达水平显著增高;与DGC组比较,Shh蛋白IGC表达水平显著降低.各组间比较均有显著统计学差异(P<0.001).β-catenin在3组之间的表达有统计学差异(F=253.540,P<0.001).与CSG组比较,β-catenin蛋白在IGC组表达水平显著增高,在DGC组表达水平显著降低;与DGC组比较,β-catenin蛋白在IGC组表达水平显著增高,各组间比较均有显著统计学差异(P<0.001).Shh与β-catenin在IGC与DGC中表达呈负相关(P<0.001).Wnt3A与Sfrp1在3组中多不表达.结论:从CSG到IGC发展进程中,Shh减少,可能打破了Shh与Wnt两个信号通路在胃腺体中的动态平衡,β-catenin表达水平增高,且部分从细胞膜转移至细胞核表达,激活了经典Wnt通路,导致了IGC发生;从CSG到DGC发展进程中,Shh表达显著增多,可能与DGC发病密切相关.
Objective: To observe the expression characteristics of sonic hedgedog(Shh), Wnt3A/β-catenin and secreted frizzled-related protein-1(Sfrp1) in chronic superficial gastritis(CSG), intestinal type gastric cancer(IGC) and diffuse type gastric cancer(DGC), and to study their correlation, thus to preliminarily reveal the completely different pathogenesis of these two types of gastric cancer. Methods: The expression of Shh, Wnt3A/β-catenin and Sfrp1 in 30 cases of CSG(control group), 42 cases of IGC and 42 cases of DGC was detected by immunohistochemical method and Western Blotting method. The difference of expression of each protein in each group was compared and the correlation between them was analyzed. Results: The results of immunohistochemical test showed that Shh was mainly expressed in cytoplasm and cell membrane. In CSG, Shh was mostly positively or strongly positively expressed. In IGC, expression of Shh decreased to be weakly positive expression or even no expression in some cases. In DGC, Shh exhibited mostly strong positive expression. The expression of Shh was significantly different among the three groups(H=46.064, P<0.001). The expression of Shh in CSG was significantly higher than that in IGC(P<0.001), but significantly lower than that in DGC(P<0.05). The expression level of Shh in DGC was significantly higher than that in IGC(P<0.001). In CSG, β-catenin is mainly expressed in cell membrane. The cytoplasm is generally not stained or locally light stained, and the nucleus is not stained. In IGC, the expression of β-catenin was significantly increased, and the ectopic expression in cytoplasm and nucleus was observed. But the expression was significantly decreased in DGC and even not observed in some cases. The expression of β-catenin was significantly different among the three groups(H=38.629, P<0.001).The expression level of β-catenin in CSG was significantly lower than that in IGC(P<0.05),but significantly higher than that in DGC(P<0.001). The expression of β-catenin in IGC was significantly higher than that in DGC(P<0.001). The results of Western Blotting test indicated that the expression of Shh was significantly different among the three groups(F=739.731, P<0.001). Compared with CSG group, the expression level of Shh in IGC group decreased markedly, but increased significantly in DGC group. Compared with DGC group, the expression level of Shh in IGC group decreased significantly. There was significant statistical difference among the groups(P<0.001). The expression of β-catenin was significantly different among the three groups(F=253.540, P<0.001). Compared with CSG group, the expression level of β-catenin in IGC group increased significantly, but decreased significantly in DGC group. Compared with DGC group, the expression level of β-catenin in IGC group increased significantly. There was significant difference among groups(P<0.001). The expression of Shh and β-catenin was negatively correlated in IGC and DGC(P<0.001). Wnt3A and Sfrp1 were mostly not expressed in the three groups. Conclusion:In the development from CSG to IGC, the decrease of Shh may break the dynamic balance between Shh and Wnt signaling pathway in gastric glands. The expression level of β-catenin increased, and its expression was partly transferred from cell membrane to nucleus, which activates the classical Wnt pathway and leads to IGC. In the development from CSG to DGC, the significant increase of Shh expression may be closely related to the pathogenesis of DGC.
Objective: To observe the expression characteristics of sonic hedgedog(Shh), Wnt3A/β-catenin and secreted frizzled-related protein-1(Sfrp1) in chronic superficial gastritis(CSG), intestinal type gastric cancer(IGC) and diffuse type gastric cancer(DGC), and to study their correlation, thus to preliminarily reveal the completely different pathogenesis of these two types of gastric cancer. Methods: The expression of Shh, Wnt3A/β-catenin and Sfrp1 in 30 cases of CSG(control group), 42 cases of IGC and 42 cases of DGC was detected by immunohistochemical method and Western Blotting method. The difference of expression of each protein in each group was compared and the correlation between them was analyzed. Results: The results of immunohistochemical test showed that Shh was mainly expressed in cytoplasm and cell membrane. In CSG, Shh was mostly positively or strongly positively expressed. In IGC, expression of Shh decreased to be weakly positive expression or even no expression in some cases. In DGC, Shh exhibited mostly strong positive expression. The expression of Shh was significantly different among the three groups(H=46.064, P<0.001). The expression of Shh in CSG was significantly higher than that in IGC(P<0.001), but significantly lower than that in DGC(P<0.05). The expression level of Shh in DGC was significantly higher than that in IGC(P<0.001). In CSG, β-catenin is mainly expressed in cell membrane. The cytoplasm is generally not stained or locally light stained, and the nucleus is not stained. In IGC, the expression of β-catenin was significantly increased, and the ectopic expression in cytoplasm and nucleus was observed. But the expression was significantly decreased in DGC and even not observed in some cases. The expression of β-catenin was significantly different among the three groups(H=38.629, P<0.001).The expression level of β-catenin in CSG was significantly lower than that in IGC(P<0.05),but significantly higher than that in DGC(P<0.001). The expression of β-catenin in IGC was significantly higher than that in DGC(P<0.001). The results of Western Blotting test indicated that the expression of Shh was significantly different among the three groups(F=739.731, P<0.001). Compared with CSG group, the expression level of Shh in IGC group decreased markedly, but increased significantly in DGC group. Compared with DGC group, the expression level of Shh in IGC group decreased significantly. There was significant statistical difference among the groups(P<0.001). The expression of β-catenin was significantly different among the three groups(F=253.540, P<0.001). Compared with CSG group, the expression level of β-catenin in IGC group increased significantly, but decreased significantly in DGC group. Compared with DGC group, the expression level of β-catenin in IGC group increased significantly. There was significant difference among groups(P<0.001). The expression of Shh and β-catenin was negatively correlated in IGC and DGC(P<0.001). Wnt3A and Sfrp1 were mostly not expressed in the three groups. Conclusion:In the development from CSG to IGC, the decrease of Shh may break the dynamic balance between Shh and Wnt signaling pathway in gastric glands. The expression level of β-catenin increased, and its expression was partly transferred from cell membrane to nucleus, which activates the classical Wnt pathway and leads to IGC. In the development from CSG to DGC, the significant increase of Shh expression may be closely related to the pathogenesis of DGC.
引文
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[18] KONSTANTINOU D,BERTAUX-SKEIRIK N,ZAVROS Y.Hedgehog signaling in the stomach[J].Curr Opin Pharmacol,2016,31:76-82.
[19] SONG X,XIN N,WANG W,et al.Wnt/β-catenin,an oncogenic pathway targeted by H.pylori in gastric carcinogenesis[J].Oncotarget,2015,6(34):35579-35588.
[20] CHIURILLO M A.Role of the Wnt/β-catenin pathway in gastric cancer:An in-depth literature review[J].World J Exp Med,2015,5(2):84-102.
[21] EBERT M P,FEI G,KAHMANN S,et al.Increased beta-catenin mRNA levels and mutational alterations of the APC and beta-catenin gene are present in intestinal-type gastric cancer[J].Carcinogenesis,2002,23(1):87-91.
[22] XIAO C,OGLE S A,SCHUMACHER M A,et al.Hedgehog signaling regulates E-cadherin expression for the maintenance of the actin cytoskeleton and tight junctions[J].Am J Physiol Gastrointest Liver Physiol,2010,299(6):G1252-1265.
[23] DING M,WANG X.Antagonism between Hedgehog and Wnt signaling pathways regulates tumorigenicity[J].Oncol Lett,2017,14(6):6327-6333.
[24] HAI B,YANG Z,MILLAR S E,et al.Wnt/β-catenin signaling regulates postnatal development and regeneration of the salivary gland[J].Stem Cells Dev,2010,19(11):1793-1801.
[2] CORREA′S P.Human gastric carcinogenesis:a multistep and multifactorial process-First American Cancer Society Award lecture on cancer epidemiology and prevention[J].Cancer Res,1992,52(24):6735-6740.
[3] SONG H,EKHEDEN I G,ZHENG Z,et al.Incidence of gastric cancer among patients with gastric precancerous lesions:observational cohort study in a low risk Western population[J].BMJ,2015,351:h3867.
[4] ANSARI S,GANTUYA B,TUAN V P,et al.Diffuse gastric cancer:A summary of analogous contributing factors for its molecular pathogenicity[J].Int J Mol Sci,2018,19(8):2424.
[5] WESSLER S,KRISCH L M,ELMER D P,et al.From inflammation to gastric cancer-the importance of Hedgehog/GLI signaling in Helicobacter pylori-induced chronic inflammatory and neoplastic diseases[J].Cell Commun Signal.2017,15(1):15.
[6] YANAI K,NAKAMURA M,AKIYOSHI T,et al.Crosstalk of hedgehog and Wnt pathways in gastric cancer[J].Cancer Lett,2008,263(1):145-156.
[7] MORRIS S L,HUANG S.Crosstalk of the Wnt/β-catenin pathway with other pathways in cancer cells[J].Genes Dis,2016,3(1):41-47.
[8] CARBALLO G B,HONORATO J R,de LOPES G P F,et al.A highlight on Sonic hedgehog pathway[J].Cell Commun Signal,2018,16(1):11.
[9] WANG L,TAN C,QIAO F,et al.Upregulated expression of DIXDC1 in intestinal-type gastric carcinoma:co-localization with β-catenin and correlation with poor prognosis[J].Cancer Cell Int,2015,15(1):120.
[10] AKYALA A I,PEPPELENBOSCH M P.Gastric cancer and Hedgehog signaling pathway:emerging new paradigms[J].Genes Cancer,2018,9(1/2):1-10.
[11] 刘先勇,刘雪梅,杨振斌.附子理中汤加味治疗慢性萎缩性胃炎肠上皮化生的临床疗效及机制[J].云南中医学院学报,2016,39(4):54-57.LIU X Y,LIU X M,YANG Z B.Clinical curative effects on modified fuzi lizhong decoction on chronic atrophic gastritis with intestinal metaplasia and its mechanism[J].Journal of Yunnan University of Traditional Chinese Medicine,2016,39(4):54-57.
[12] 刘先勇,王建宁.Hedgehog与胃腺体分化及Correa级联学说关系的研究进展[J].中国细胞生物学学报,2015,37(11):1572-1580.LIU X Y,WANG J N.Progress in studies of relationship of hedgehog and the gastric gland differentiation,correas cascade[J].Chinese Journal of Cell Biology,2015,37(11):1572-1580.
[13] 赵坤,张翠萍,张琪,等.SHH在肠型胃癌发展中的表达及意义[J].齐鲁医学杂志,2012,27(3):214-216.ZHAO K,ZHANG C P,ZHANG Q,et al.The expression of SHH and its significance in the development of intestinal-type gastric cancer[J].Medical Journal of Qilu,2012,27(3):214-216.
[14] LEE S Y,HAN H S,LEE K Y,et al.Sonic hedgehog expression in gastric cancer and gastric adenoma[J].Oncol Rep,2007,17(5):1051-1055.
[15] FUKAYA M,ISOHATA N,OHTA H,et al.Hedgehog signal activation in gastric pit cell and in diffuse-type gastric cancer[J].Gastroenterology,2006,131(1):14-29.
[16] RIQUELME I,SAAVEDRA K,ESPINOZA J A,et al.Molecular classification of gastric cancer:Towards a pathway-driven targeted therapy[J].Oncotarget,2015,6(28):24750-24779.
[17] OHTA H,AOYAGI K,FUKAYA M,et al.Cross talk between hedgehog and epithelial-mesenchymal transition pathways in gastric pit cells and in diffuse-type gastric cancers[J].Br J Cancer,2008,100(2):389-398.
[18] KONSTANTINOU D,BERTAUX-SKEIRIK N,ZAVROS Y.Hedgehog signaling in the stomach[J].Curr Opin Pharmacol,2016,31:76-82.
[19] SONG X,XIN N,WANG W,et al.Wnt/β-catenin,an oncogenic pathway targeted by H.pylori in gastric carcinogenesis[J].Oncotarget,2015,6(34):35579-35588.
[20] CHIURILLO M A.Role of the Wnt/β-catenin pathway in gastric cancer:An in-depth literature review[J].World J Exp Med,2015,5(2):84-102.
[21] EBERT M P,FEI G,KAHMANN S,et al.Increased beta-catenin mRNA levels and mutational alterations of the APC and beta-catenin gene are present in intestinal-type gastric cancer[J].Carcinogenesis,2002,23(1):87-91.
[22] XIAO C,OGLE S A,SCHUMACHER M A,et al.Hedgehog signaling regulates E-cadherin expression for the maintenance of the actin cytoskeleton and tight junctions[J].Am J Physiol Gastrointest Liver Physiol,2010,299(6):G1252-1265.
[23] DING M,WANG X.Antagonism between Hedgehog and Wnt signaling pathways regulates tumorigenicity[J].Oncol Lett,2017,14(6):6327-6333.
[24] HAI B,YANG Z,MILLAR S E,et al.Wnt/β-catenin signaling regulates postnatal development and regeneration of the salivary gland[J].Stem Cells Dev,2010,19(11):1793-1801.