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Shc1在肝癌组织中的表达及对肝癌SMMC-7721细胞增殖迁移能力的影响
戴敏1,刘俊1,朱海燕2,顾澄宇3,陶汉川3,乔谦3,杨树东4,蔡兵3*
0
(1. 南通大学附属海安医院普通外科, 南通 226600;
2. 南通大学附属海安医院肿瘤科, 南通 226600;
3. 南京医科大学附属无锡市人民医院肝胆外科, 无锡 214000;
4. 南京医科大学附属无锡市人民医院病理科, 无锡 214000*通信作者)
摘要:
目的 探讨Shc1在肝癌组织中的表达情况及其对人肝癌SMMC-7721细胞增殖、迁移能力的影响。方法 采用实时定量PCR、蛋白质印迹法、免疫组织化学法检测33例肝癌及其对应的癌旁组织中Shc1的表达情况,并分析Shc1蛋白表达量与临床特征间的关系;培养肝癌SMMC-7721细胞,以siRNA技术干扰Shc1的表达,采用MTT法和细胞划痕试验分别检测干扰Shc1对细胞增殖活力和迁移能力的影响。结果 在33对样本中,肝癌组织中Shc1的表达高于癌旁组织(P<0.05);Shc1表达于肝癌细胞的胞质中。无肝硬化的患者Shc1阳性率(100%)高于伴肝硬化者(54.2%),术前Child-Pugh分级A级的患者Shc1阳性率(75.9%)高于B级或C级的患者(0%),术前血AFP阳性患者Shc1阳性率(79.2%)高于AFP阴性患者(33.3%),术后病理Edmondson Ⅲ、Ⅳ级患者Shc1阳性率(84.2%)高于Ⅰ、Ⅱ级的患者(42.9%),差异均有统计学意义(P<0.05)。干扰Shc1表达后,随时间延长,SMMC-7721细胞增殖、迁移明显受到抑制(P<0.05)。结论 Shc1在肝癌组织中高表达,且Shc1的阳性率与有无肝硬化、Child-Pugh分级、术前AFP、病理Edmondson分级相关;干扰Shc1的表达可抑制肝癌SMMC-7721细胞的增殖、迁移能力。
关键词:  Shc1  肝细胞癌  细胞增殖  细胞迁移  RNA干扰
DOI:10.3724/SP.J.1008.2014.00944
投稿时间:2013-11-26修订日期:2014-04-30
基金项目:国家自然科学基金青年项目(81302105),无锡市科技计划项目(CYE00917).
Expression of Shc1 in hepatocellular carcinoma tissues and its effect on proliferation and migration of human hepatocarcinoma SMMC-7721 cells
DAI Min1,LIU Jun1,ZHU Hai-yan2,GU Cheng-yu3,TAO Han-chuan3,QIAO Qian3,YANG Shu-dong4,CAI Bing3*
(1. Department of General Surgery, Haian Hospital of Nantong University, Nantong 226600, Jiangsu, China;
2. Department of Oncology, Haian Hospital of Nantong University, Nantong 226600, Jiangsu, China;
3. Department of Hepatobiliary Surgery, Wuxi People's Hospital, Nanjing Medical University, Wuxi 214000, Jiangsu, China;
4. Department of Pathology, Wuxi People's Hospital, Nanjing Medical University, Wuxi 214000, Jiangsu, China
* Corresponding author.)
Abstract:
Objective To investigate the expression of Shc1 in hepatocellular carcinoma tissues and its effect on the proliferation and migration of human hepatocarcinoma SMMC-7721 cells. Methods The expression of Shc1 in 33 hepatocellular carcinoma tissues and corresponding adjacent tissues was examined by real-time PCR, Western blotting analysis and immunohistochemistry method; the relationship between the expression and clinical features was also analyzed. Shc1 was silenced by RNA interference technique in SMMC-7721 cells to determine its effect on proliferation and migration; MTT assay and wound healing assay were used to examine the effect of Shc1 knockdown on cell vitality and migration. Results Real-time PCR, Western blotting analysis and immunohistochemistry demonstrated that Shc1 expression in hepatocellular carcinoma tissues was significantly higher than that in the adjacent tissues (P<0.05); the expression of Shc1 was mainly in the cytoplasma. Shc1 positive rates in the patients without cirrhosis (100%), of Child-Pugh class A before operation (75.9%), AFP-positive before operation (79.2%) and pathology Edmondson Ⅲ/Ⅳ level after operation (84.2%) were significantly higher than those with cirrhosis (54.2%), of Child-Pugh class B or C (0%), AFP-negative (33.3%), and pathology Edmondson Ⅰ/Ⅱ(42.9%, P all <0.05). Shc1 expression silence resulted in significant decrease in the proliferation and migration of SMMC-7721 cells (P<0.05). Conclusion Shc1 is overexpressed in hepatocarcinoma tissue. The expression of Shc1 is associated with cirrhosis, Child-Pugh class, preoperation AFP and pathology Edmondson grades of the hepatocarcinoma. Inhibition of Shc1 expression can inhibit the proliferation and migration of human hepatocarcinoma cell SMMC-7721 in vitro.
Key words:  Shc1  hepatocellular carcinoma  cell proliferation  cell migration  RNA interference