靶向干扰TAGLN表达对HBV阳性肝癌细胞生物学行为的影响及机制初探

目的:探究TAGLN对HBV阳性肝癌细胞HepG2. 2. 15生物学行为的影响及可能的作用机制。方法:免疫组化法和Western blot检测TAGLN在HBV阳性和HBV阴性肝癌组织及细胞中的表达差异;用TAGLN干扰慢病毒感染HepG2. 2. 15细胞,通过嘌呤霉素筛选干扰TAGLN表达的稳定表达细胞系,Western blot验证干扰效率; CCK-8法和克隆形成实验检测干扰TAGLN表达对HepG2. 2. 15细胞增殖能力的影响; Transwell实验检测干扰TAGLN表达对HepG2. 2. 15细胞迁移和侵袭的影响; Western blot检测PI3K、p-PI3K、AKT以及p-AKT的表达。结果:TAGLN在HBV阳性肝癌组织及细胞中的表达高于HBV阴性肝癌组织和细胞(P 0. 01);干扰TAGLN表达能抑制HepG2. 2. 15细胞增殖、克隆形成能力、迁移和侵袭(P 0. 01);降低HepG2. 2. 15细胞中PI3K和AKT(P 0. 01)及p-PI3K和p-AKT(P 0. 05)的表达。结论:在肝癌组织中,HBV感染能增加TAGLN的表达;干扰TAGLN表达后HepG2. 2. 15细胞的增殖能力、克隆形成能力、迁移和侵袭的能力减弱,其机制可能与PI3K及AKT的表达减少有关。

Effect of Targeted Interference with TAGLN Expression on Biological Behavior of HBV-Positive Hepatocellular Carcinoma Cells and Its Mechanisms

Objective: To invest igate the effect of TAGLN on the biological behavior of HBV-positive hepatocellular carcinoma cell line HepG2.2.15 and its underlying mechanism.Methods: Immunohistochemistry and Western blot were used to detect the expression levels of TAGLN in HBV-positive and HBV-negative hepatocellular carcinoma tissues and cells; HepG2.2.15 cells were infected with TAGLN interfering lentiviruses, and stable expression cell lines were screened by purinomycin. Then transfection efficiency was confirmed by Western blot; CCK-8 and clone formation assay were used to detect the proliferation of HepG2.2.15 cells. While cell migration and invasion capabilities were determined by Transwell assay. The expression levels of PI3K, p-PI3K, AKT and p-AKT were examined by Western blot.Results: The expression of TAGLN in HBV-positive hepatocellular carcinoma tissues and cells was higher than that in HBV-negative hepatocellular carcinoma tissues and cells(P<0.01); Knockdown of TAGLN could effectively inhibit cell proliferation, migration and invasion (P<0.01), as well as down-regulate the expression of PI3K, AKT(P<0.01), p-PI3K and p-AKT (P<0.05).Conclusion: HBV infection can increase the expression of TAGLN in hepatocellular carcinoma. After interfering with TAGLN expression, the proliferation, cloning, migration and invasion of HepG2.2.15 cells decreased, which may be related to the decrease of PI3K and AKT expression.