国际肿瘤学杂志

国际肿瘤学杂志 ›› 2015, Vol. 42 ›› Issue (8): 580-584.doi: 10.3760/cma.j.issn.1673422X.2015.08.006

• 论著 • 上一篇    下一篇

SH2B在原发性肝癌中的表达及其分子机制研究

华建江, 唐发清, 段朝军, 袁咏梅, 何亚, 陈望, 汪奇云   

  1. 518104 广州医科大学附属深圳沙井医院检验科(华建江、袁咏梅、何亚、陈望、汪奇云)；暨南大学附属珠海医院检验科(唐发清)；中南大学湘雅医院医学实验研究中心(段朝军)
  • 出版日期:2015-08-08 发布日期:2015-06-29
  • 通讯作者: 华建江，Email: hjianjiang13@163.com E-mail:hjianjiang13@163.com
  • 基金资助:

    深圳市科技计划（201203292）

The expression and molecular mechanisms of SH2B in hepatocarcinoma

HUA  Jian-Jiang, TANG  Fa-Qing, DUAN  Chao-Jun, YUAN  Yong-Mei, HE  Ya, CHEN  Wang, WANG  Qi-Yun   

  1. Department of Clinical Laboratory, Shenzhen Shajing Affiliated Hospital of Guangzhou Medical University, Shenzhen 518104, China
  • Online:2015-08-08 Published:2015-06-29
  • Contact: Hua Jianjiang E-mail:hjianjiang13@163.com

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摘要： 目的分析接头蛋白SH2B在原发性肝癌中的表达,探讨其在肝癌癌变中的分子机制。 方法用免疫组织化学SABC法检测SH2B在27例肝炎、29例肝硬化、47例肝癌患者组织中的表达。免疫荧光筛选低表达SH2B的肝癌细胞HepG2，设转染组（转染pcDNA3.1SH2B质粒）、空载体组（转染pcDNA3.1空白载体）和对照组（未转染）。Western blotting检测基因转染效率，MTT法分析细胞增殖情况，集落形成试验分析其对细胞集落形成的影响，流式细胞术分析细胞周期变化。 结果SH2B在肝癌患者组织中的表达阳性率为95.7%，明显高于肝硬化组的55.2%（χ2=18.64, P＜0.01）和肝炎组的25.9%（χ2=40.01, P＜0.01）。肝癌细胞HepG2转染pcDNA3.1SH2B质粒后可获得SH2B有效表达；培养48 h后转染组肝癌HepG2细胞平均吸光度（A）值为1.12±0.19，明显高于空载体组的0.45±0.11（t=-31.55, P＜0.01)，提示SH2B促进肝癌细胞增殖；转染组细胞集落数为166±14，明显高于空载体组的82±8（t=-20.33, P＜0.01）和对照组的78±9（t=-19.64, P＜0.01），提示SH2B显著促进肝癌细胞HepG2细胞集落形成；转染组S期细胞比例为(45.7±5.8)%，明显高于空载体组的(19.4±4.7)%（t=-20.33, P＜0.01)和对照组的(20.5±5.1)%（t=-34.69, P＜0.01），提示SH2B促进肝癌HepG2细胞周期演进。 结论SH2B在肝癌中高表达，可能通过促进人体内肝癌细胞的细胞周期演进、增殖及转化参与肝癌的癌变过程。

关键词: 肝肿瘤, SH2B, 分子机制

Abstract: ObjectiveTo observe the expression and influence of SH2B in hepatocarcinoma, and to investigate the molecular mechanisms of canceration in hepatocarcinoma. MethodsBy using SABC imunohistochemistry, the expressions of SH2B were detected in 27 cases of hepatitis, 29 cases of hepatocirrhosis and 47 cases of hepatocarcinoma. Hepatocarcinoma cell (HepG) 2 with a lowexpressed SH2B was selected using immunofluorescence assay. There were 3 groups: the transfected group (transfected with pcDNA3.1SH2B), the vector group (transfected with pcDNA3.1) and the blank group (without transfection). After gene transfection, SH2B expression was detected by Western blotting; cell proliferation was measured by MTT assay; cell colony was counted by colony formation test; and cell cycle was analyzed by flowcy tometer. ResultsThe positive rate of SH2B in hepatocarcinoma (95.7%) was significantly higher than 55.2% in hepatocirrhosis (χ2=18.64, P＜0.01) and 25.9% in hepatitis (χ2=40.01, P＜0.01). After being transfected with pcDNA 3.1SH2B, SH2B expression dramatically increased in HepG2 cells. After cultured for 48 h, the average optical density value of the transfected group was 1.12±0.19, obviously higher than 0.45±0.11 in the vector group (t=-31.55, P＜0.01), which indicated that cells proliferation was significantly enhanced after being transfected with SH2B. The cell colony numbers of the transfected group was 166±14, significantly higher than 82±8 in the vector group (t=-20.33, P＜0.01) and 78±9 in the blank group (t=-19.64, P＜0.01), which indicated that the cell colony numbers increased after being transfected with SH2B. The S stage cells of the transfected group was (45.7±5.8)%, significantly higher than (19.4±4.7)% in the vector group (t=-20.33, P＜0.01) and (20.5±5.1)% in the blank group (t=-34.69, P＜0.01), which indicated that SH2B could enhance promote cell cycle of HepG2 cells. ConclusionThe expression of SH2B in hepatocarcinoma is high, and it may be involved in the canceration of hepatocarcinoma though promoting cell cycle, cell proliferation and cell transformation.

Key words: Liver neoplasms, SH2B, Molecular mechanisms