Zhang Yan, Hu Mei-yu, Chen Bi-hua, Wang Zhi-jun, Wu Wei-zhong, Zhou Kang, Liu Kang-da
Experimental Research, Zhongshan Hospital, Fudan University, Shanghai 200032, China.
Zhonghua Gan Zang Bing Za Zhi. 2006 Jul;14(7):489-94.
To investigate the different expressions of cytoskeletal organizer ezrin and cytoskeleton protein beta- and gamma-actin in hepatocellular carcinoma (HCC) cell lines with different metastatic potentials and to explore the role of ezrin in cell growth and metastasis in HCC cell lines SF7721 and MHCC97-H.
Immunofluorescence, RT-PCR and Western blot were used to detect the gene and protein expressions of ezrin and actin in hepatocellular carcinoma cell lines with different metastatic potentials. RNA interference (RNAi) was applied to down-regulate the ezrin expression in SF7721 and MHCC97-H. Changes of the cell growth and metastasis potentials after the RNAi treatment were studied. MTT assay was used to detect cell proliferation changes and Transwell assay was applied to observe the changes of cell motility and invasiveness.
Both ezrin and cytoskeleton protein were demonstrated in the cytoplasma of the cells at the same time. The expression of them in cell lines with high metastatic potential, such as SF7721, MHCC-1 and MHCC97-H was obviously higher than in those with low metastatic potentials, such as SMMC-7721, Hep3B and HepG2 (chi2= 13.277, P = 0.010; chi2= 21.815). The mRNA and ezrin and cytoskeleton protein gamma-actin were over-expressed in HCC cell lines with high metastatic potentials. The expressions of beta-actin of cell lines with different metastatic potentials showed no differences. Ezrin protein was successfully down-regulated and the proliferation and the invasiveness of the cells decreased with low ezrin protein level in SF7721 and MHCC97-H.
Over-expression of ezrin and cytoskeleton protein gamma-actin are associated with the process of metastasis of hepatocellular carcinoma cells. The growth and invasiveness of SF7721 and MHCC97-H cells can be inhibited by down-regulating ezrin expression.
研究细胞骨架组织者埃兹蛋白(ezrin)以及细胞骨架蛋白β-肌动蛋白和γ-肌动蛋白在具有不同转移潜能的肝癌细胞系中的不同表达情况,并探讨埃兹蛋白在肝癌细胞系SF7721和MHCC97-H细胞生长和转移中的作用。
采用免疫荧光法、逆转录-聚合酶链反应(RT-PCR)和蛋白质免疫印迹法检测不同转移潜能肝癌细胞系中埃兹蛋白和肌动蛋白的基因及蛋白表达。应用RNA干扰(RNAi)技术下调SF7721和MHCC97-H细胞中埃兹蛋白的表达。研究RNAi处理后细胞生长和转移潜能的变化。采用噻唑蓝(MTT)法检测细胞增殖变化,应用Transwell法观察细胞运动和侵袭能力的变化。
埃兹蛋白和细胞骨架蛋白均同时在细胞胞质中被证实存在。它们在高转移潜能细胞系如SF7721、MHCC-1和MHCC97-H中的表达明显高于低转移潜能细胞系如SMMC-7721、Hep3B和HepG2(χ2 = 13.277,P = 0.010;χ2 = 21.815)。高转移潜能肝癌细胞系中埃兹蛋白mRNA以及细胞骨架蛋白γ-肌动蛋白呈过表达。不同转移潜能细胞系中β-肌动蛋白的表达无差异。在SF7721和MHCC97-H细胞中,埃兹蛋白被成功下调,随着埃兹蛋白水平降低,细胞增殖和侵袭能力下降。
埃兹蛋白和细胞骨架蛋白γ-肌动蛋白的过表达与肝癌细胞的转移过程相关。下调埃兹蛋白表达可抑制SF7721和MHCC97-H细胞的生长和侵袭能力。