黏蛋白16对皮革胃恶性表型的影响  

Effect of mucin 16 on the malignant phenotype of gastric linitis plastica

作　　者：张歆雨 刘铸 洪莲莲[2,3] 凌志强 Zhang Xinyu;Liu Zhu;Hong Lianlian;Ling Zhiqiang(Department of Oncology,the Second Clinical Medical College of Zhejiang Chinese Medicine University,Hangzhou 310053,China;Zhejiang Cancer Institute,Zhejiang Cancer Hospital,Hangzhou 310022,China;Hangzhou Institute of Medicine Chinese Academy of Sciences,Hangzhou 310018,China)

机构地区：[1]浙江中医药大学第二临床医学院肿瘤科,杭州310053 [2]浙江省肿瘤研究所,浙江省肿瘤医院,杭州310022 [3]中国科学院杭州医学研究所,杭州310018

出　　处：《中华实验外科杂志》2024年第5期974-978,共5页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金面上项目(32271238);浙江省自然科学基金面上项目(LY20H160005);浙江省卫生领军人才(浙卫办[2021]40号);浙江省公益技术研究计划实验动物项目(LGD20H160003);浙江省基础公益研究计划项目(LGF21H160010);国家卫生健康委科学研究基金-浙江省卫生健康重大科技计划重点项目(WKJ-ZJ-2117)。

摘　　要：目的:观察黏蛋白16(MUC16)基因在皮革胃中的表达和MUC16对胃癌细胞的增殖、侵袭等生物学行为的影响。方法:收集2009年1月至2017年12月在浙江省肿瘤医院首次经病理诊断的13例皮革胃组织标本,用免疫组织化学染色法检测MUC16的表达。采用荧光定量聚合酶链反应(PCR)和蛋白质印迹法(Western blot)分析MUC16的表达。选取MUC16高表达的细胞系,利用慢病毒构建MUC16敲低的稳转细胞株。将实验分为MUC16敲低组(MUC16-KO组)和阴性对照(NC组),采用细胞计数试剂盒(CCK-8)和克隆形成实验检测不同细胞的增殖能力,用划痕愈合实验和Transwell实验检测不同细胞的迁移和侵袭能力。定性资料用χ^(2)检验,定量资料采用独立样本t检验或单因素方差分析结果。结果:皮革胃组织样本中MUC16的蛋白表达水平明显高于常见胃癌组织(2.99±0.30比1.07±0.48),t=5.942,P<0.05)。人胃癌细胞系KATOⅢ、NUGC4、HGC27和AGS中MUC16的信使RNA(messenger RNA,mRNA)的表达水平均高于人正常胃上皮细胞系GES1(9.93±0.79、8.69±0.54、6.37±0.61、3.58±3.30比1.00±0.10,t=19.368、24.373、15.045、13.848,均P<0.05)。人弥漫型胃癌细胞系KATOⅢ、NUGC4、HGC27中MUC16的mRNA表达水平均高于人胃腺癌细胞系AGS(9.93±0.79、8.69±0.54、6.37±0.61比3.58±0.30,t=12.959、14.331、7.082,均P<0.05)。人胃癌细胞系KATOⅢ、NUGC4、HGC27和AGS中MUC16的蛋白表达水平均高于人正常胃上皮细胞系GES1(0.79±0.01、0.62±0.02、0.53±0.00、0.38±0.01比0.08±0.00,t=122.969、46.765、202.339、36.109,均P<0.05)。且人弥漫型胃癌细胞系KATOⅢ、NUGC4、HGC27中的MUC16蛋白表达水平高于胃腺癌细胞AGS(0.79±0.01、0.62±0.02、0.53±0.00比0.38±0.01,t=50.215、18.590、23.986,均P<0.05)。两种细胞株中的MUC16-KO组在96 h时间点吸光度值均低于NC组[AGS细胞株(0.86±0.02比1.21±0.42),t=12.973,P<0.05;HGC27细胞株(0.45±0.30比1.04±0.11),t=8.936,P<0.05]。两种细胞株MUC16-KO组�Objective To investigate the expression of mucin16(MUC16)gene in gastric linitis plastica and the effects of MUC16 on the proliferation and invasion of gastric cancer cells.Methods In total,13 cases of gastric linitis plastica tissue samples from January 2009 to December 2017 in Zhejiang Cancer Hospital were collected and the expression of MUC16 was detected by immunohistochemical staining.Fluorescence quantitative polymerase chain reaction(PCR)and Western blotting were used to analyze the expression of MUC16.Cells with high MUC16 expression were selected and lentivirus was used to construct stable cell lines with low MUC16 knockdown.The experiment was divided into MUC16 knockdown group(MUC16-KO group)and negative control group(NC group).Cell counting kit-8(CCK-8)and clonal formation assays were used to detect the proliferation ability of different cells,and scratch healing assay and Transwell assay were used to detect the migration and invasion ability of different cells.Chi-square test was used for qualitative data,and independent sample t-test or One-way analysis of variance was used for quantitative data.Results The expression level of MUC16 protein in leather stomach tissue samples was significantly higher than that in common gastric cancer tissues(2.99±0.30 vs.1.07±0.48,t=5.942,P<0.05).The mRNA expression of MUC16 in human gastric cancer cell lines KATOⅢ,NUGC4,HGC27 and AGS was higher than that of human normal gastric epithelial cell line GES1(9.93±0.79,8.69±0.54,6.37±0.61,3.58±3.30 vs.1.00±0.10,t=19.368,24.373,15.045,13.848,P<0.05).The mRNA expression levels of MUC16 in human diffuse gastric cancer cell lines KATOⅢ,NUGC4 and HGC27 were higher than those in human gastric adenocarcinoma cell line AGS(9.93±0.79,8.69±0.54,6.37±0.61 vs.3.58±0.30,t=12.959,14.331,7.082,P<0.05).The expression levels of MUC16 in human gastric cancer cell lines KATOⅢ,NUGC4,HGC27 and AGS were higher than those in human normal gastric epithelial cell lines GES1(0.79±0.01,0.62±0.02,0.53±0.00 and 0.38±0.01 vs.0.08±

关 键 词：胃癌 增殖 转移 侵袭 

分 类 号：R735.2[医药卫生—肿瘤]