TGF-β 通过磷酸化 smad3 连接区诱导胃癌中 fascin 的表达。
TGF-β induces fascin expression in gastric cancer via phosphorylation of smad3 linker area.
机构信息
Department of Pathology, Xiangya Hospital, Central South University Changsha, China ; Department of Pathology, Xiangya Medical School, Central South University Changsha, China.
出版信息
Am J Cancer Res. 2015 May 15;5(6):1890-6. eCollection 2015.
BACKGROUND
Fascin is an actin-bundling protein critical for tumor invasion. TGF-β could induce fascin expression in gastric cancer cells. In this study, we attempted to explore the role of p-smad3L in the expression of fascin induced by TGF-β in gastric cancer cells.
METHODS
Pseudopodia were evaluated by immunofluorescence. Fascin expression was detected by RT-PCR and western blot. Smad3 siRNA was used to repress the endogenous smad3. The phosphorylations of smad3 linker region at sites s204, s208 and s213 were detected by western blot. The fascin promoter reporter activity was measured by dual luciferase assay.
RESULTS
TGF-β could increase the formation of pseudopodia and the expression of fascin in gastric cancer cells. Smad3 depletion abrogated the expression of fascin induced by TGF-β. The phosphorylation of smad3 linker region at serine 204, 208 and 213 was enhanced in gastric cancer cells after TGF-β treatment. The fascin promoter reporter activity was significantly enhanced with TGF-β treatment in both wild-type Smad3 group and Smad3EPSM group (P<0.05). Furthermore, the fascin promoter reporter activity in the wild-type Smad3 transfectant cells was significantly higher than that in Smad3EPSM cells (P<0.05).
CONCLUSIONS
fascin expression induced by TGF-β depends on smad3, at least in part, depends on smad3 linker phosphorylation.
背景
Fascin 是一种肌动蛋白束状蛋白，对肿瘤侵袭至关重要。TGF-β可诱导胃癌细胞中 fascin 的表达。在这项研究中，我们试图探讨 p-smad3L 在 TGF-β诱导的胃癌细胞中 fascin 表达中的作用。
方法
通过免疫荧光评估伪足。通过 RT-PCR 和 Western blot 检测 fascin 的表达。用 Smad3 siRNA 抑制内源性 Smad3。通过 Western blot 检测 Smad3 连接区位点 s204、s208 和 s213 的磷酸化。通过双荧光素酶测定法测量 fascin 启动子报告活性。
结果
TGF-β可增加胃癌细胞伪足的形成和 fascin 的表达。Smad3 耗竭可阻断 TGF-β诱导的 fascin 表达。TGF-β处理后，胃癌细胞中 Smad3 连接区丝氨酸 204、208 和 213 的磷酸化增强。TGF-β处理后，野生型 Smad3 组和 Smad3EPSM 组的 fascin 启动子报告活性均显著增强（P<0.05）。此外，野生型 Smad3 转染细胞的 fascin 启动子报告活性明显高于 Smad3EPSM 细胞（P<0.05）。
结论
TGF-β诱导的 fascin 表达至少部分依赖于 smad3，依赖于 smad3 连接区磷酸化。
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