Department of Anti-Aging Medicine, The University of Tokyo, Tokyo, Japan.
Int J Cancer. 2012 May 15;130(10):2240-8. doi: 10.1002/ijc.26224. Epub 2011 Sep 22.
ADP ribosylation factor GTPase-activating protein 3 (ARFGAP3) is a GTPase-activating protein that associates with the Golgi apparatus and regulates the vesicular trafficking pathway. In the present study, we examined the contribution of ARFGAP3 to prostate cancer cell biology. We showed that ARFGAP3 expression was induced by 100 nM of dihydrotestosterone (DHT) at both the mRNA and protein levels in androgen-sensitive LNCaP cells. We generated stable transfectants of LNCaP cells with FLAG-tagged ARFGAP3 or a control empty vector and showed that ARFGAP3 overexpression promoted cell proliferation and migration compared with control cells. We found that ARFGAP3 interacted with paxillin, a focal adhesion adaptor protein that is important for cell mobility and migration. Small interfering RNA (siRNA)-mediated knockdown of ARFGAP3 showed that ARFGAP3 siRNA markedly reduced LNCaP cell growth. Androgen receptor (AR)-dependent transactivation activity on prostate-specific antigen (PSA) enhancer was synergistically promoted by exogenous ARFGAP3 and paxillin expression, as shown by luciferase assay in LNCaP cells. Thus, our results suggest that ARFGAP3 is a novel androgen-regulated gene that can promote prostate cancer cell proliferation and migration in collaboration with paxillin.
ADP 核糖基化因子 GTP 酶激活蛋白 3（ARFGAP3）是一种与高尔基体相关并调节囊泡运输途径的 GTP 酶激活蛋白。在本研究中，我们研究了 ARFGAP3 对前列腺癌细胞生物学的贡献。结果显示，100 nM 的二氢睾酮（DHT）可诱导雄激素敏感的 LNCaP 细胞中 ARFGAP3 的 mRNA 和蛋白水平表达。我们通过转染 FLAG 标记的 ARFGAP3 或对照空载体生成稳定的 LNCaP 细胞系，结果显示 ARFGAP3 过表达与对照细胞相比促进细胞增殖和迁移。我们发现 ARFGAP3 与粘着斑衔接蛋白 paxillin 相互作用，paxillin 是一种对细胞迁移和运动很重要的黏着斑衔接蛋白。小干扰 RNA（siRNA）介导的 ARFGAP3 敲低表明，ARFGAP3 siRNA 可显著降低 LNCaP 细胞的生长。通过 LNCaP 细胞中的 luciferase 检测，我们发现外源性 ARFGAP3 和 paxillin 的表达协同促进了雄激素受体（AR）对前列腺特异性抗原（PSA）增强子的依赖性转录激活活性。因此，我们的结果表明 ARFGAP3 是一种新型的雄激素调节基因，可与 paxillin 协同促进前列腺癌细胞的增殖和迁移。
