肝细胞核因子4γ在胃癌组织中的表达及其对胃癌细胞增殖和干性的影响  

Expression of hepatocyte nuclear factor 4γin gastric carcinoma and its role in cell proliferation and stemness

作　　者：贾永旭[1] 汤虹 常志伟[1] 樊慧杰[1] 关新元[3] 秦艳茹[1] Jia Yongxu;Tang Hong;Chang Zhiwei;Fan Huijie;Guan Xinyuan;Qin Yanru(Department of Oncology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China;Department of Oncology,Cancer Hospital Affiliated Zhengzhou University,Zhengzhou 450003,China;Department of Clinical Oncology,the University of Hong Kong,Hong Kong 999077,China)

机构地区：[1]郑州大学第一附属医院肿瘤内科,450000 [2]郑州大学附属肿瘤医院内科,450003 [3]香港大学李嘉诚医学院临床肿瘤系,999077

出　　处：《中华肿瘤杂志》2021年第11期1164-1169,共6页Chinese Journal of Oncology

基　　金：国家自然科学基金面上项目(81472605)。

摘　　要：目的探讨肝细胞核因子4γ(HNF4γ)在胃癌细胞增殖和干性维持中的作用及机制。方法收集2012—2015年在郑州大学第一附属医院行胃癌根治术患者的胃癌及相应癌旁正常组织蜡块102例,新鲜组织42例。分别采用实时荧光定量聚合酶链反应、逆转录聚合酶链反应和免疫组化法检测HNF4γ的表达情况。建立HNF4γ过表达胃癌细胞株AGS-HNF4γ和HNF4γ沉默细胞株HGC27-shHNF4γ,分别采用XTT细胞增殖、平板克隆实验、干细胞成球实验测定HNF4γ过表达和沉默对胃癌细胞增殖和干性的影响,Western blot检测CD44的表达。结果 42例新鲜胃癌组织中,HNF4γ mRNA的表达水平为12.43±2.702,高于相应癌旁正常组织3.639±1.109(P<0.010)。102例石蜡包埋胃癌组织中,41.2%(42/102)HNF4γ蛋白表达阳性,高于癌旁正常组织的8.8%(9/102,P<0.001)。HNF4γ蛋白表达与胃癌患者的分化程度、浸润深度、临床分期和淋巴结转有关(均P<0.05)。HNF4γ蛋白高表达组患者的中位生存时间为25个月,3年生存率为4.8%(2/42),HNF4γ蛋白正常表达组患者的中位生存时间为38个月,3年生存率为51.7%(31/60),差异均有统计学意义(均P<0.001)。AGS-HNF4γ细胞增殖快于对照AGS-Vector细胞,细胞克隆数高于AGS-Vector细胞[分别为(243.5±24.5)个和(81.0±16.0)个,P=0.030],细胞成球率高于AGS-Vector细胞[分别为(83.5±3.9)%和(21.8±5.6)%,P=0.010],CD44的表达量高于AGS-Vector细胞。HGC27-shHNF4细胞增殖慢于对照HGC27-Vector细胞,细胞克隆数低于HGC27-Vector细胞[分别为(26.0±1.0)个和(83.5±4.5)个,P=0.006],细胞成球率低于HGC27-Vector细胞[分别为(20.8±8.4)%和(72.5±4.8)%,P=0.030],CD44的表达量低于HGC27-Vector细胞。结论 HNF4γ在胃癌组织中异常高表达,与胃癌患者的不良预后有关。HNF4γ可促进胃癌细胞的增殖,并通过促进CD44的表达维持胃癌细胞的干性。Objective To explore the role and molecular mechanism of hepatocyte nuclear factor 4γ(HNF4γ)in proliferation and stemness of gastric cancer.Methods A total of 102 cases of paraffin-embedded gastric cancer tissues and matched adjacent gastric tissues and 42 cases of fresh-frozen tissues derived from gastric patients who received radical gastrectomy were collected from the First Affiliated Hospital of Zhengzhou University between 2012 to 2015.The expression of HNF4γwas tested by immunohistochemical staining,quantitative real-time polymerase chain reaction(qRT-PCR).HNF4γoverexpressed(AGS-HNF4γ)and shRNA silenced(HGC27-shHNF4γ)gastric cell lines were established.The effects of HNF4γon cell proliferation and stemness were verified by XTT,clone formation and sphere formation assay.The expression of CD44 was detected by western blot.Results The mRNA expression level of HNF4γin fresh-frozen gastric cancer tissue was(12.43±2.702),which was significantly higher than(3.639±1.109)in normal tissue(P<0.001).The high protein expression rate of HNF4γin paraffin-embedded gastric cancer tissues was 41.2%(42/102),which was significantly higher than 8.8%(9/102)in normal gastric mucosa tissue(P<0.001).The protein expression of HNF4γwas closely related to the tumor differentiation,infiltration depth,lymph node metastasis and tumor stage(P<0.05).The median survival interval of patients with HNF4γhigh expression was 25 months,the 3-year survival rate was 4.8%(2/42),significantly lower than 38 months and 51.7%(31/60)of patients with normal HNF4γexpression(P<0.001).The proliferation and CD44 protein expression of AGS-HNF4γcells were significantly higher than those of the AGS-Vector cells.The number of clone formation,sphere formation rate of AGS-HNF4γcells were 243.5±24.5 and(83.5±3.9)%,significantly higher than 81.0±16.0 and(21.8±5.6)%of AGS-Vector cells(P=0.030 and P=0.010,respectively).The proliferation and CD44 protein expression of HGC27-shHNF4 cells were significantly lower than those of the HGC27-vector cells.Th

关 键 词：胃肿瘤 肝细胞核因子 预后 增殖 肿瘤干细胞 

分 类 号：R735.2[医药卫生—肿瘤]