特异性siRNA沉默LASP-1对HepG2细胞增殖和迁移的影响

目的应用RNA干扰技术沉默细胞内LASP-1蛋白的表达，探讨LASP-1蛋白对人肝癌HepG2细胞增殖和迁移的作用。方法根据LASP-1mRNA编码序列设计RNA干扰靶点并构建特异性小干扰RNA（siRNA），通过脂质体转染入HepG2细胞，采用Westernblot法检测LASP-1蛋白的表达。采用CCK-8增殖实验、平板克隆形成实验、Transwell和划痕愈合实验方法，观察HepG2细胞增殖及迁移情况。结果（1）LASP-1siRNA对LASP．1蛋白表达水平有显著的下调作用（P〈0．05）。（2）体外实验结果显示，与HepG2组及阴性对照组相比，siRNA组HepG2细胞增殖和克隆形成率明显被抑制（P〈0．05），其迁移能力也下降（P〈0．05）。结论下调LASP-1蛋白表达可显著抑制HepG2细胞的增殖和迁移能力，提示LASP-1在肝癌细胞恶性增殖和转移过程中具有重要作用，为临床上寻找抑制肝癌增殖转移的靶点提供新的思路和方法。

Effect of silencing LASP-1 expression on HepG2 cell proliferation and migration by specific small interfering RNA

Objective To investigate the effect of LASP-1 expression silencing by specific small in- terfering RNA （siRNA） on HepG2 cell proliferation and migration. Methods small interfering RNA （siRNA） targeting LASP-1 were designed and transfected into HepG2 cells by lipofectamine. Western blotting was used to evaluate the effect of siRNA. When the expression level of LASP-1 was downregulated after successful transfection, cell proliferation was detected by CCK-8 assay and the plate colony formation assay. The migration ability of cells was detected by transwell assay and wound healing assay. Results （ 1 ） Western blotting results revealed that LASP-1 siRNA notably downregulated the expression of LASP- 1 protein （P 〈 0.05 ）. （2） Downregulation of LASP-1 expression distinctly inhibited the proliferation of the HepG2 cells （ P 〈 0.05 ）. The migration ability of HepG2 cells in vitro was decreased significantly af- ter siRNA targeting LASP-1 was successful transfected （ P 〈 0.05 ）. Conclusion LASP-1 might play an important role in HepG2 cell proliferation and migraion, indicating that LASP-1 may be a novel and effec- tive therapeutic target for preventing hepatocellular carcinoma proliferation and migration.