Fermitin家族同源蛋白1在胆囊癌中的表达及其在胆囊癌细胞增殖与侵袭中的作用  

Expression level and pathologic function of fermitin family homolog 1 in gallbladder cancer development

作　　者：韩旭晟 李雪川 邹路 崔旭雅 李怀峰[1,2] 张飞 张一鉴[1,2] 龚伟 李茂岚 Han Xusheng;Li Xuechuan;Zou Lu;Cui Xuya;Li Huaifeng;Zhang Fei;Zhang Yijian;Gong Wei;Li Maolan(Department of General Surgery,Xinhua Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200092,China;Shanghai Key Laboratory of Biliary Tract Disease Research,Shanghai 200092,China;Department of Biliary-Pancreatic Surgery,Renji Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200127,China)

机构地区：[1]上海交通大学医学院附属新华医院普外科,200092 [2]上海市胆道疾病研究重点实验室,200092 [3]上海交通大学医学院附属仁济医院胆胰外科,200127

出　　处：《中华实验外科杂志》2022年第5期869-872,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金面上项目(82073206、81773043)。

摘　　要：目的检测Fermitin家族同源蛋白1(FERMT1)在胆囊癌患者组织样本中的表达水平并分析其与患者临床预后的关系,探讨FERMT1对胆囊癌细胞生物学行为的影响。方法回顾性分析上海交通大学医学院附属新华医院2016年6月至2018年12月共30例胆囊癌患者的临床资料,采用门诊及电话的方式对患者进行随访,随访时间截止至2020年12月。利用免疫组织化学染色法检测30例胆囊癌患者癌及其癌旁组织中FERMT1蛋白的表达,并统计分析其与患者临床病理特征及预后的关系。通过小干扰RNA敲减胆囊癌细胞NOZ、GBC-SD中的FERMT1蛋白表达,分别将胆囊癌细胞分为FERMT1敲减组(si-FERMT1)和阴性对照组(si-NC),利用细胞计数试剂盒(CCK-8)检测各组细胞的增殖活性,Transwell法检测细胞侵袭能力,蛋白质印迹法(Western blot)测定上皮-间充质转化(EMT)标志蛋白的表达水平,两组间比较采用t检验。结果免疫组织化学结果显示,胆囊癌组织中FERMT1蛋白表达阳性率(33.00%,9/30)高于癌旁组织(3.33%,1/30,P<0.05),组织样本中FERMT1的表达与患者TNM分期、淋巴结转移密切相关(P<0.05)。FERMT1蛋白高表达组中位生存时间低于低表达组(14个月比18个月,Log-rank=3.875,P<0.05)。CCK-8实验结果显示转染96 h后,NOZ细胞中si-FERMT1组吸光度值为低于si-NC组(0.773±0.046比1.124±0.043,t=12.395,P<0.01);GBC-SD细胞中si-FERMT1组吸光度值为低于si-NC组(0.538±0.022比0.778±0.033,t=13.457,P<0.01)。Transwell实验结果显示,NOZ细胞中si-FERMT1组侵袭细胞数低于si-NC组[(178.00±15.72)个比(430.33±20.60)个,t=16.874,P<0.01];GBC-SD细胞中si-FERMT1组侵袭细胞数低于si-NC组[(130.33±11.72)个比(414.67±20.53)个,t=20.842,P<0.01],差异均有统计学意义。结论FERMT1在胆囊癌组织中呈高表达,并与胆囊癌发展及不良预后明显相关,在胆囊癌形成及发展过程中发挥促癌作用。Objective To investigate the expression of fermitin family homolog 1(FERMT1)protein in tissue samples of patients with gallbladder carcinoma and its relationship with clinical prognosis,and the pathologic impact of FERMT1 on proliferation and invasion of gallbladder cancer(GBC)cells.Methods The clinical data of 30 patients with gallbladder cancer admitted in Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine from June 2016 to December 2018 were retrospectively analyzed.The patients were followed-up by outpatient examination and telephone interview until December 2020.The expression level of FERMT1 protein in tumor tissues and adjacent tissues of the 30 patients with gallbladder cancer was detected by immunohistochemistry and its relation with clinical characteristics and prognosis was analyzed.The FERMT1 gene in gallbladder cancer cells NOZ and GBC-SD was knocked down by small interference RNA.Gallbladder cancer cells were divided into FERMT1 knockdown group(si-FERMT1)and negative control group(si-NC).Cell proliferation activity was detected by cell counting kit-8(CCK-8)assay.Cell invasion ability was evaluated by Transwell method.The expression levels of epithelial-mesenchymal transition(EMT)markers were detected by Western blotting.T-test was used between the two groups.Results The positive rate of FERMT1 protein expression in gallbladder carcinoma(33.00%,9/30)was higher than that in paracancerous tissues(3.33%,1/30,P<0.05).The expression of FERMT1 in tissue samples was closely related to TNM stage and lymph node metastasis(P<0.05).The median survival of GBC patients expressing higher FERMT1 was 14 month compared to 18 months observed in patients expressing lower FERMT1(Log-rank=3.875,P<0.05).The CCK-8 assay indicated that at 96 h after transfection,the absorbance values of si-FERMT1 group in NOZ cells was 0.773±0.046,which was significantly lower than in the si-NC group(1.124±0.043,t=12.395,P<0.01).The absorbance values of the si-FERMT1 group in GBC-SD cells was 0.538±0.022,which

关 键 词：胆囊癌 Fermitin家族同源蛋白1 增殖 侵袭 

分 类 号：R735.8[医药卫生—肿瘤]