Role of hepatitis B virus X protein in regulating LIM and SH3 protein 1 (LASP-1) expression to mediate proliferation and migration of hepatoma cells.
Role of hepatitis B virus X protein in regulating LIM and SH3 protein 1 (LASP-1) expression to mediate proliferation and migration of hepatoma cells.
复制标题
DOI：
10.1186/1743-422x-9-163
复制
发表时间：
2012-08-16
期刊：
影响因子：
4.8
通讯作者：
Zheng K
中科院分区：
文献类型：
--
作者：
Tang R;Kong F;Hu L;You H;Zhang P;Du W;Zheng K
乙型肝炎病毒 X 蛋白 (HBx) 已被证明与乙型肝炎病毒感染引起的肝细胞癌 (HCC) 的发生有关。然而，其对肝细胞癌进展的潜在影响仍不清楚。 LIM 和 SH3 蛋白 1 (LASP-1)（一种粘着斑蛋白）在 HCC 组织中以上调方式表达。 LASP-1在HCC增殖和迁移的调控中发挥着重要作用。在本研究中，我们研究了 LASP-1 在 HBx 相关肿瘤进展中的作用。通过RT-PCR和蛋白质印迹分析检测HBx稳定转染的HepG2和Huh-7细胞中LASP-1的水平。通过免疫荧光分析评估 LASP-1 的细胞定位。通过蛋白质印迹测定证明了磷脂酰肌醇 3-激酶 (PI3-K) 途径的活性。用针对 LASP-1 的特异性小干扰 RNA (siRNA) 转染表达 HBx 的细胞。通过细胞活力测定和平板克隆形成测定评估细胞的增殖和迁移能力。通过Transwell实验和伤口愈合实验检测细胞的迁移能力。 RT-PCR和蛋白质印迹分析表明，与对照细胞相比，稳定表达HBx的细胞中LASP-1的表达增加。免疫荧光研究显示HepG2-HBX细胞中LASP-1主要分布在伪足和细胞质中，而HepG2-Mock细胞中LASP-1主要分布在细胞质中。 Huh-7-HBX 细胞中 LASP-1 的细胞定位位于核周部分，而 Huh-7-Mock 细胞中 LASP-1 主要定位于细胞质。用 PI3-K 通路抑制剂 LY294002 治疗后，LASP-1 的上调受到抑制。在稳定的HBx表达细胞中过表达LASP-1增强了肝细胞的增殖和迁移能力。 siRNA介导的LASP-1在稳定表达HBx的细胞中的表达显着抑制肝细胞的增殖和迁移。这些结果表明，HBx可以通过PI3-K途径上调LASP-1，促进肝癌细胞的增殖和迁移。
Hepatitis B virus X protein (HBx) has been shown to be responsible for the development of hepatocellular carcinoma (HCC) caused by Hepatitis B virus infection. However, its potential effect on the progression of hepatocellular carcinoma remains yet unclear. LIM and SH3 protein 1 (LASP-1), a focal adhesion protein, is expressed in an up-regulation manner in the HCC tissues. LASP-1 plays an important role in the regulation of proliferation and migration of HCC. In this study, we investigated the effect of LASP-1 involved in HBx-related tumor progression. LASP-1 levels in the HBx stable transfected HepG2 and Huh-7 cells were detected by RT-PCR and western blot analysis. The cellular localization of LASP-1 was assessed by immunofluorescence analysis. The activity of phosphatidylinositol 3-kinase (PI3-K) pathway was demonstrated by western blot assay. The HBx-expressing cells were transfected with specific small interference RNA (siRNA) against LASP-1. The proliferation and migration ability of cells were evaluated by cell viability assay and plate clone formation assay. The migration ability of cells was detected by transwell assay and wound healing assay. RT-PCR and western blot analysis indicated the expression of LASP-1 was increased in the stable HBx-expressing cells compared with the control cells. Immunofluorescence study revealed that the distributions of LASP-1 in HepG2-HBX cells were mainly in pseudopods and the cytoplasm while they were mainly localized in the cytoplasm of HepG2-Mock cells. The cellular localizations of LASP-1 in Huh-7-HBX cells were in the perinuclear fractions while they were mainly localized in the cytoplasm of Huh-7-Mock cells. The upregulation of LASP-1 was inhibited after treatment with LY294002, PI3-K pathway inhibitor. Overexpression of LASP-1 in the stable HBx-expressing cells enhanced the proliferation and migration ability of hepatocellular cells. siRNA-mediated LASP-1 knowdown in the stable HBx-expressing cells significantly suppressed hepatocellular cells proliferation and migration. These results demonstrated that HBx could upregulate LASP-1 through PI3-K pathway to promote the proliferation and migration of hepatoma cells.