shRNA下调Tspan8基因对肝癌细胞侵袭与转移的影响

The influence of down-regulation of Tspan 8 by shRNA on metastasis and invasion of hepatocellular carcinomas

目的探讨Tspan8基因对人肝癌细胞侵袭与转移的影响。方法应用RT-PCR与Western印迹检测不同转移潜能肝癌细胞及组织中Tspan8的表达；应用shRNA—Tspan8（Lv／GFP／Tspan8）下调Tspan8表达；MTT法检测细胞增殖变化；Western印迹检测基质金属蛋白酶-12表达改变；最后应用Transwell检测细胞侵袭能力变化。结果高表达Tspan8见于高转移潜能的肝癌细胞，Tspan8在肝癌组织中表达明显高于相应癌旁组织。感染Lv／GFP／Tspan8、LV／GFP后，LV／GFP／Tspan8感染肝癌细胞的Tspan8表达显著下调（P〈0．01）；下调Tspan8后，细胞增殖反应变化不明显（24h，48h与72h，P〉0．05）；但MMP-12表达抑制明显（P〈O．01），细胞侵袭能力明显下降。结论下调Tspan8基因的表达对肝癌侵袭与转移能力有明显抑制。

Objective To study the effect of Tspan 8 on metastasis and invasion of human hepa- tocellular carcinomas（HCC）. Methods RT-PCR and western blot were used to detect the expressions of Tspan 8 in HCC cell lines, HCC and matched nontumorous tissues. The expression of Tspan 8 was then down-regulated by LV/GFP/Tspan 8 in HCC cells. The expressions of Tspan 8 mRNA and protein were determined by RT-PCR and Western blot assay, respectively. The proliferation was examined by MTT, the expression of AMDM12 was assessed by Western blot, and the invasion ability of HCC cells was evaluated by transwells. Results A high level of Tspan 8 was found in high metastatic potential HCC cells, and the expression of Tspan 8 in HCC tissues was much higher than that in the matched nontumorous tissues. Down-regulation of Tspan 8 had no influence on the proliferation of HCC cells （P〉0. 05）, while it inhibited the expression of ADAM12 and the invasive ability of HCC cells （P〈0.01, P〈0. 01 respectively）. Conclusion Tspan 8 played an important role in invasion and metastasis of human hepatocellular carcinomas and down-regulation by LV/GFP/Tspan 8 inhibited the invasiveness of human hepatocellular carcinoma cells.