摘要
目的音猬因子(sonic hedgehog,Shh)与Wnt3A/β-catenin在Correa级联反应中发生了深刻变化,与肠型胃癌(intestinal-type gastric cancer,IGC)的发病密切相关。本研究观察Shh与Wnt3A/β-catenin在IGC的Correa级联反应不同病理阶段中表达特点,揭示IGC重要发病分子生物学机制。方法选取2016-01-01-2018-12-31南京医科大学附属江宁医院内镜室和病理科获得的40例慢性浅表性胃炎(chronic superficial gastritis,CSG)、40例慢性萎缩性胃炎(chronic atrophic gastritis,CAG)伴肠上皮化生(intestinal metaplasia,IM)及40例IGC标本,运用免疫组织化学法及蛋白质印迹法检测各组标本Shh和Wnt3A/β-catenin蛋白表达水平,并分析相关性。结果 CSG组Shh主要表达在细胞浆和细胞膜,着色"+、++及+++"阳性率分别为10%、62.50%和27.50%;CAG伴IM组Shh阳性率分别为55%、35%和10%;IGC组Shh阳性率分别为25%、55%、10%和10%。3组Shh表达差异有统计学意义,H=37.335,P<0.001。CAG伴IM组和IGC组Shh表达水平低于CSG组,Z值分别为-4.055和-5.569,均P<0.001;IGC组Shh表达水平低于CAG伴IM组,Z=-3.083,P=0.002。CSG组β-catenin主要表达在细胞膜,着色阳性率97.5%;细胞核一般不表达,着色阳性率5%;CAG伴IM组和IGC组,β-catenin在细胞膜表达降低,着色阳性率分别为80%和82.5%,在细胞核表达增强,着色阳性率分别是27.5%和57.5%。3组β-catenin在细胞膜(H=10.937,P=0.004)和细胞核(H=31.306,P<0.001)表达差异有统计学意义。CAG伴IM组及IGC组细胞膜上β-catenin表达水平低于CSG组,Z值分别为-2.871和-2.834,P值分别为0.004和0.005;但两组细胞核上β-catenin表达水平高于CSG组,Z值分别为-2.722和-5.135,均P<0.05;β-catenin在IGC组细胞核中表达水平高于CAG伴IM组,Z=-3.533,P<0.001。Wnt3A在3组均不表达。蛋白质印迹法结果显示,从CSG、CAG伴IM发展至IGC过程中,Shh表达水平逐渐降低,β-catenin表达逐渐增加。3组中Shh蛋白相对表达量分别为1.00±0.08、0.51±0.09和0.14±0.11,差异有统计学意义,F=176.758,P<0.001;其中CAG伴IM组和IGC组Shh表达较CSG组减少,IGC组Shh表达较CAG伴IM组减少,差异均有统计学意义,均P<0.001。3组中β-catenin蛋白相对表达量分别为0.61±0.07、1.01±0.06和1.70±0.17,差异有统计学意义,F=71.711,P<0.001;其中CAG伴IM组和IGC组β-catenin表达较CSG组增多,IGC组β-catenin表达较CAG伴IM组增多,均P<0.05。在CSG中,Shh表达水平与β-catenin在细胞膜和细胞核表达水平呈负相关,r值分别为-0.288和-0.930,均P<0.05。在CAG伴IM中,Shh表达水平与β-catenin在细胞膜和细胞核表达水平亦呈负相关,r值分别为-0.221和-0.762,均P<0.05。结论在Correa级联反应中,伴随CSG、CAG伴IM进展到IGC,Shh逐渐减少,可能打破了Shh与Wnt两个信号通路动态平衡,β-catenin从细胞膜逐渐转移至细胞核,激活了经典的Wnt信号通路是IGC发病的重要的分子机制。
OBJECTIVE Sonic hedgehog(Shh)and Wnt3 A/beta catenin have undergone profound changes in Correa cascade,which is closely related to the pathogenesis of intestinal-type gastric cancer(IGC).The aim of the study was to observe the expression of Shh and Wnt3 A/β-catenin in the different pathological stages of Correa’s cascade of IGC,and to reveal the important molecular biological mechanism of IGC.METHODS Forty cases of chronic superficial gastritis(CSG),40 cases of chronic atrophic gastritis(CAG)with intestinal metaplasia(IM)and 40 cases of IGC were selected from Endoscopy Room and Pathology Department of Jiangning Hospital Affiliated to Nanjing Medical University from January 1 st,2016 to December 31 st,2018.Immunohistochemical method and Western Blotting method were used to detect the protein expression levels of Shh and Wnt3 A/β-catenin RESULTS In CSG group,Shh is mainly expressed in cytoplasm and cell membrane,the positive rates of Shh staining"+,++and+++"were 10%,62.50%and 27.50%,respectively.In CAG with IM group,the positive rates of Shh staining were 55%,35%and 10%,respectively.In IGC group,the positive rates of Shh staining were 25%,55%,10%and 10%respectively.There was significant difference in the expression of Shh among the three groups,H=37.335,P<0.001.The expression level of Shh in CAG with IM group and IGC group was significantly lower than that in CSG group,Zvalue were-4.055 and-5.569,respectively,all P<0.001.The expression level of Shh in IGC group was significantly lower than that in CAG with IM group,P=0.002.In CSG group,β-catenin was mainly expressed in the cell membrane,and the positive staining rate was 97.5%,but most of them were not expressed in the nucleus,and the staining positive rate was 5%.In CAG with IM group and IGC group,the expression ofβ-catenin was decreased in the cell membrane,the positive rates of staining were 80%and 82.5%respectively,and the expression ofβ-catenin in the nucleus was enhanced,the positive rate of staining was 27.5%and 57.5%,respectively.The expression ofβ-catenin in cell membrane(H=10.937,P=0.004)and nucleus(H=31.306,P<0.001)of the three groups had significant difference.The expression level ofβ-catenin in CAG with IM group and IGC group was lower than that in CSG group,Zvalue was-2.871 and-2.834,Pvalue was 0.004 and 0.005,respectively,but the expression level ofβ-catenin in nucleus of two groups was higher than that in CSG group,Zvalue was-2.722 and-5.135,respectively,all P<0.05.The expression level ofβ-catenin in the nucleus of IGC group was higher than that of CAG with IM group,Z=-3.533,P<0.001.Wnt3 Awas not expressed among the three groups.During the development from CSG and CAG with IM to IGC,the expression level of Shh gradually decreased and the expression level ofβ-catenin gradually increased.The relative expression levels of Shh protein in the three groups were 1.00±0.08,0.51±0.09 and 0.14±0.11,respectively.The difference was statistically significant(F=176.758,P<0.001).Shh expression in CAG with IM group and IGC group was lower than that in CSG group,and Shh expression in IGC group was lower than that in CAG with IM group,and the difference was statistically significant(all P<0.001).The relative expression levels ofβ-catenin in the three groups were 0.61±0.07,1.01±0.06 and 1.70±0.17,respectively,The difference was statistically significant(F=71.711,P<0.001).The expression ofβ-catenin in CAG with IM group and IGC group was higher than CSG group,and the expression ofβ-catenin in IGC group was higher than CAG with IM group,all P<0.05.In CSG,the expression level of Shh was negatively correlated with that ofβ-catenin in cell membrane and nucleus,and the rvalues were 0.288 and 0.930,respectively,all P<0.05.In CAG with IM,the expression level of Shh was negatively correlated with the expression level ofβ-catenin in cell membrane and nucleus,with r values of-0.221 and-0.762,all P<0.05.CONCLUSIONS In Correa’s cascade reaction,with the progress of CSG,CAG and IM to IGC,Shh decreases gradually,which may break the dynamic balance between Shh and Wnt signaling pathways.β-catenin gradually transfers from cell membrane to nucleus,activating the classical Wnt signaling pathway is an important molecular mechanism of IGC.
作者
刘先勇
王玉红
张若梅
杨进
孟春芹
王建宁
仇伟
李加虎
周宇
LIU Xian-yong;WANG Yu-hong;ZHANG Ruo-mei;YANG Jin;MENG Chun-qin;WANG Jian-ning;QIU Wei;LI Jia-hu;ZHOU Yu(Jiangning Hospital,Nanjing Medical University,Nanjing 211100,P.R.China)
出处
《中华肿瘤防治杂志》
CAS
北大核心
2019年第20期1525-1530,共6页
Chinese Journal of Cancer Prevention and Treatment
基金
南京市卫生计生委医学科技发展项目（YKK16234）
作者简介
第一作者:刘先勇,男,河南商城人,硕士,主任医师,主要从事中西医结合防治胃癌前病变的研究工作。Tel:86-25-52281848 E-mail:liuxianyongl973@163.com;通讯作者:王建宁,男,江苏南京人,主任医师,主要从事胃癌的临床与基础研究工作。Tel:86-25-51190501 E-mail:Jianghuail973@163.com。
