氧化应激诱导的SHP1/Src失衡对晚期胃食管结合部腺癌侵袭生长的影响  

SHP1/Src imbalance induced by oxidative stress promotes the invasion and growth of advanced adenocarcinoma of esophagogastric junction

作　　者：范彩霞 贾慧 范艳明 王志恒 丁晓玲[2] FAN Caixia;JIA Hui;FAN Yanming;WANG Zhiheng;DING Xiaoling(Linfen Central Hospital,Linfen 041000,Shanxi,China;People’s Hospital of Ningxia Hui Autonomous Region,Yinchuan 750011,Ningxia,China)

机构地区：[1]临汾市中心医院,山西临汾041000 [2]宁夏回族自治区人民医院,宁夏银川750011

出　　处：《现代中西医结合杂志》2024年第21期2946-2952,共7页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：宁夏自然科学基金项目(2021AAC03299)。

摘　　要：目的探讨氧化应激诱导的含SH2结构域的蛋白酪氨酸磷酸酶1(SHP1)/酪氨酸激酶(Src)失衡对晚期胃食管结合部腺癌(AEG)侵袭生长的影响。方法①选取宁夏回族自治区人民医院2020年1月—2023年4月手术治疗的100例AEG患者,根据肿瘤TNM分期分为早期AEG组和晚期AEG组,HE染色观察癌组织病理形态,免疫组化法观察癌组织中磷酸化Src(p-Src)蛋白阳性表达情况,Western blot法检测癌组织中p-Src、烟酰胺腺嘌呤二核苷酸磷酸氧化酶4(NOX4)和烟酰胺腺嘌呤二核苷酸磷酸氧化酶2(NOX2)蛋白表达情况。②通过体外培养人AEG肿瘤细胞,CCK-8法检测肿瘤细胞增殖能力,Transwell实验检测肿瘤细胞迁移能力,Western blot法检测细胞中p-SHP1、p-Src、磷酸化信号传导和转录激活因子3(p-STAT3)、磷酸化RUNX家族转录因子3(p-RUNX3)及RUNX3蛋白表达情况。结果与早期AEG组比较,晚期AEG组癌组织出现明显缺血缺氧坏死,癌组织中p-Src蛋白阳性表达率和p-Src、NOX2和NOX4蛋白相对表达量均明显升高(P均<0.05)。体外实验中,与对照组比较,NAC组培养72 h细胞光密度值明显降低(P均<0.05),细胞迁移数目明显减少(P<0.05),细胞中p-SHP1、RUNX3蛋白相对表达量均明显升高(P均<0.05),细胞中p-Src、p-STAT3、p-RUNX3蛋白相对表达量均明显降低(P均<0.05);H_(2)O_(2)组培养72 h细胞光密度值明显升高(P<0.05),细胞迁移数目明显增加(P<0.05),细胞中p-SHP1、RUNX3蛋白相对表达量均明显降低(P均<0.05),细胞中p-Src、p-STAT3、p-RUNX3蛋白相对表达量均明显升高(P均<0.05)。与H_(2)O_(2)组比较,H_(2)O_(2)+NAC组、H_(2)O_(2)+达沙替尼组、H_(2)O_(2)+达沙替尼+NAC组培养72 h细胞光密度值明显降低(P均<0.05),细胞迁移数目均明显减少(P均<0.05),细胞中RUNX3蛋白相对表达量均明显升高(P均<0.05),细胞中p-SHP1、p-Src、p-STAT3、p-RUNX3蛋白相对表达量均明显降低(P均<0.05)。H_(2)O_(2)+达沙替尼组、H_(2)O_(2)+达沙替尼+NObjective It is to investigate the effect of oxidative stress(OS)-induced imbalance of SH2 domain-containing protein tyrosine phosphatase 1(SHP1)/tyrosine kinase(Src)on the invasion and growth of advanced adenocarcinoma of esophagogastric junction(AEG).Methods①One hundred cases of patients with AEG treated with surgery in People’s Hospital of Ningxia Hui Autonomous Region from January 2020 to April 2023 were selected and divided into early AEG group and late AEG group according to TNM stage.The pathological morphology of cancer tissues was observed by HE staining,the positive protein expression of phosphorylated Src(p-Src)in cancer tissues was detected by immunohistochemistry,and the protein expressions of p-Src,NADPH oxidase 4(NOX4),and NADPH oxidase 2(NOX2)in cancer tissues were detected by Western blot assay.②Human AEG tumor cells were cultured in vitro,and their proliferation ability was detected by CCK-8 assay,their migration ability was detected by Transwell assay,and the protein expressions of p-SHP1,p-Src,phosphorylated signal transducer and activator of transcription 3(p-STAT3),phosphorylated RUNX family transcription factor 3(p RUNX3)and RUNX3 were detected by Western blot assay.Results Compared with the early AEG group,the cancer tissues in the advanced AEG group was with obvious ischemia-hypoxia necrosis,and the positive expression rate of p-Src protein and the relative expressions of p-Src,NOX2 and NOX4 proteins in the cancer tissues were significantly increased(all P<0.05).In the in vitro experiments,compared with the control group,the optical density value of cells cultured for 72 h in the NAC group was significantly decreased(P<0.05),the number of migrated cells was significantly reduced(P<0.05),the relative expressions of p-SHP1 and RUNX3 proteins in the cells were significantly increased(all P<0.05),and the relative expressions of p-Src,p-STAT3 and p-RUNX3 proteins in the cells were significantly decreased(all P<0.05);in the H_(2)O_(2) group,the optical density value of cells cultured for

关 键 词：胃食管结合部腺癌 氧化应激 含SH2结构域的蛋白酪氨酸磷酸酶1 酪氨酸激酶 侵袭 

分 类 号：R-332[医药卫生]