脯氨酸羟化酶-2对肝细胞肝癌生物学行为的影响及其机制  

Effect of prolyl hydroxylase domain-2 on biological behaviors of hepatocellular carcinoma and its mechanism

作　　者：李震[1] 王世豪 姚弘虞 庞春 李捷[1] Li Zhen;Wang Shihao;Yao Hongyu;Pang Chun;Li Jie(Department of Surgery,the First Affiliated Hospital of Zhengzhou University,Zhenzhou 450052,China)

机构地区：[1]郑州大学第一附属医院外科医学部,450052

出　　处：《中华实验外科杂志》2020年第2期251-254,共4页Chinese Journal of Experimental Surgery

摘　　要：目的观察脯氨酸羟化酶-2(PHD-2)对肝细胞肝癌(HCC)生物学行为的影响并探讨机制。方法应用2006年1月至2010年12月郑州大学第一附属医院HCC患者组织建立Hep3B肝癌细胞株,建立HCC裸鼠(购自北京维通利华实验动物技术有限公司)移植瘤模型,PHD-2沉默组利用瘤体内注射的方式将胆固醇修饰的PHD-2 RNA干扰(RNAi)注射入瘤体内,同时设立空白组(不做处理)和生理盐水对照组(注射生理盐水)进行对照研究,每4 d注射1次,观察并记录裸鼠的瘤体生长变化,开始注射药物后第16天处死裸鼠,采集瘤体标本用蛋白质印迹法(Western blot)技术检测PHD-2、p-Smad2/3及癌基因c-Myc的表达。采用免疫组织化学技术(IHC)检测220例HCC临床标本中c-Myc的表达,分析其与PHD-2以及HCC患者临床病理特征的相关性。计数资料进行χ^2检验,相关性分析采用Spearman等级相关检验。结果(1)在HCC裸鼠移植瘤模型中PHD-2沉默组裸鼠瘤体增长缓慢(F=15.607,P<0.05),差异有统计学意义。瘤体标本中PHD-2沉默组p-Smad2/3表达水平明显高于空白组和对照组(0.589±0.074比0.086±0.012、0.091±0.008,F=286.537,P<0.05),差异有统计学意义,而PHD-2沉默组c-Myc的表达水平明显低于其他两组(0.108±0.004比0.539±0.023、0.592±0.087,F=189.468,P<0.05),差异有统计学意义。(2)IHC技术检测220例HCC临床标本中PHD-2及c-Myc的表达,c-Myc的高表达与肿瘤体积较大、分化程度较低、TNM分期较晚、高水平的甲胎蛋白显著相关,与PHD-2的表达显著相关(r=0.557,P<0.01)。结论PHD-2的高表达可以促进HCC细胞在体内的增殖,PHD-2促进HCC进展的作用有可能是通过抑制TGF-β1-Smad信号通路中的p-Smad2/3表达,进而上调癌基因c-Myc的表达来实现的。Objective To investigate the effect of prolyl hydroxylase domain-2(PHD-2)on the biological behaviors of hepatocellular carcinoma(HCC)and to explore the related molecular mechanism.Methods HCC cell line Hep3B was selected to establish the transplantation tumor model of HCC in nude mice.The cholesterol modified PHD-2 RNA interference(RNAi)was injected into the tumor in the PHD-2 silence group.Blank group(untreated group)and normal saline group(normal saline was injected into the tumor in this group)were set up for control study.The injection was executed every 4 days and nude mice tumor growth change was recorded.The nude mice were sacrificed at 16th day after the start of the injection,and tumor tissues were collected.The expression of PHD-2,p-Smad2/3 and oncogene c-Myc was detected by Western blotting.Immunohistochemistry(IHC)technique was used to detect the expression of c-Myc in 220 cases of HCC,and the correlation between the expression of PHD-2 and clinicopathological features of patients with HCC was analyzed.Results(1)In HCC nude mice transplanted tumor model,the tumor growth in PHD-2 silence group was slow(F=15.607,P<0.05).The level of p-Smad2/3 in PHD-2 silence group was significantly higher than that in the blank group and NS control group(0.589±0.074 vs.0.086±0.012,0.091±0.008,F=286.537,P<0.05),and c-Myc level in PHD-2 silence group was significantly lower than that in the other two groups(0.108±0.004 vs.0.539±0.023,0.592±0.087,F=189.468,P<0.05).(2)The expression of c-Myc was detected in the 220 cases of clinical specimens by immunohistochemical technique.The result showed that the expression of c-Myc was significantly related with the tumor size,degree of differentiation,tumor node metastasis(TNM)stage,higher level of alpha fetoprotein(AFP),and also was significantly correlated with the expression of PHD-2(r=0.557,P<0.01).Conclusion The over-expression of PHD-2 could promote the proliferation of hepatoma cells probably by up-regulating the expression of c-Myc oncogene through inhibiting the expre

关 键 词：癌 肝细胞 脯氨酸羟化酶-2 P-SMAD2/3 C-MYC 

分 类 号：R735[医药卫生—肿瘤]