Wnt5a基因对肝癌细胞增殖及迁移能力的影响  

Wnt5a gene affects cell proliferation and migration of hepatocellular carcinoma cells

作　　者：毕利泉[1] 刘晓红[1] 陈英剑[2] 曹永成[1] 毛蕊琪[1] 耿明[1] 

机构地区：[1]济南军区总医院病理科,济南250031 [2]济南军区总医院实验诊断科,济南250031

出　　处：《临床与实验病理学杂志》2012年第8期857-861,共5页Chinese Journal of Clinical and Experimental Pathology

基　　金：山东省自然科学基金(ZR2009CM041)

摘　　要：目的探讨Wnt5a基因对人肝癌细胞MHCC97L生物学行为的影响。方法脂质体法介导Wnt5a表达及阴性对照质粒转染MHCC97L细胞;RT-PCR及Western blot法检测质粒DNA片段插入和Wnt5a蛋白的表达;借助生长曲线、平板克隆形成、细胞周期和划痕试验对转染细胞的生物学活性进行检测。结果 pcDNA3.1组细胞生长速度明显快于Wnt5a表达组细胞;pcDNA3.1组细胞克隆形成率明显高于Wnt5a表达组(P<0.05);流式细胞仪检测结果显示Wnt5a表达能抑制细胞由G1期→S期的进程;划痕试验显示,Wnt5a表达组较阴性对照组细胞迁移速度明显减慢,当加入Wnt5a抗体阻遏信号通路时,细胞迁移速度加快。结论 Wnt5a可抑制人肝癌细胞MHCC97L的增殖和迁移能力,在肿瘤生长过程中发挥肿瘤抑制基因样作用。Purpose To explore the biological effects of Wnt5a gene on hepatocellular carcinoma （HCC） cell line MHCC97L. Methods Using Lipofectamine 2000, MHCC97L cells were transfected with the pcDNA3.1-Wnt5a and the empty vector controls, respectively. The integration of the plasmid DNA and the expression of Wnt5a were confirmed by RT-PCR and Western blot, respectively. The growth curves of different cells, plate clone formation test, flow cytometry （ FCM ） and scratch asssays were used to detect the biological behaviors of the transfected cells. Results Growth curve showed that after transfection the pcDNA3.1 group cells grew faster than WntSa transfected group cells. The clone formation rates of pcDNA3.1 group was significantly higher than that of the cells with Wnt5a expression vectors transfected （ P 〈 0. 05 ）. FCM analysis showed that WntSa expression vectors transfection blocked the progression of G1-S phase in cell cycle. The result of scratch assays showed that cells transfected with WntSa expression vectors displayed lower motility than vectors control transfected cells. Blocking this pathway using antibodies to WntSa, there was an increase in cell migration. Conclusion Wnt5a expression results in inhibition of cell proliferation and invasion, suggesting that Wnt5a plays a role of a tumor suppressor gene in HCC.

关 键 词：肝肿瘤 肝细胞癌 WNT5A 

分 类 号：R735.7[医药卫生—肿瘤]