Department of Surgical Oncology, Provincial Clinical College, Fujian Medical University, Fuzhou, China.
Kaohsiung J Med Sci. 2019 Apr;35(4):214-221. doi: 10.1002/kjm2.12042. Epub 2019 Mar 21.
The present study aimed to investigate the effects of c-Ski on cell proliferation, invasion and migration of gastric cancer associated fibroblasts (CAFs). Expression of c-Ski in gastric cancer (GC) tissues was determined using immunohistochemistry. Both CAFs and non-cancerous gastric fibroblasts (NGFs) were isolated and cultured. c-Ski and Smad3 were over-expressed or knocked down using pcDNA3.0-c-Ski/Smad3 or siRNA, respectively. Cell viability, invasion and migration were measured and expression of c-Ski, α-SMA, and Smad3 in cells was determined using real time quantitative PCR (RT-qPCR) and Western blotting. Expression of c-Ski was significantly higher in both in GC tissues and cell lines, and was the highest in tissues of diffuse type. Both c-Ski and α-SMA were significantly over-expressed in CAFs compared with that in the NGFs. When c-Ski was over-expressed in NGFs, cell viability, cell invasion and migration were all enhanced and expression of Smad3 was downregulated. When c-Ski was inhibited, cell viability, cell invasion, and migration were all suppressed and expression of Smad3 was upregulated. Meanwhile, overexpression of Smad3 significantly reversed the effects of over-expressed c-Ski in NGFs, and knockdown of Smad3 dramatically reversed the effects of si-c-Ski in CAFs. Over-expressed c-Ski could enhance cell viability, promote cell invasion, and migration of GC CAFs, and the effects might be through regulation of Smad3 signaling. This study may give deeper insights for relationship between c-Ski and CAFs, as well as role of c-Ski in cancer development, and also provide some novel research targets for treatment of GC.
本研究旨在探讨 c-Ski 对胃癌相关成纤维细胞（CAFs）增殖、侵袭和迁移的影响。采用免疫组织化学法检测胃癌（GC）组织中 c-Ski 的表达。分离并培养 CAFs 和非癌性胃成纤维细胞（NGFs）。使用 pcDNA3.0-c-Ski/Smad3 或 siRNA 分别过表达或敲低 c-Ski 和 Smad3。通过实时定量 PCR（RT-qPCR）和 Western blot 检测细胞活力、侵袭和迁移，以及细胞中 c-Ski、α-SMA 和 Smad3 的表达。GC 组织和细胞系中 c-Ski 的表达均显著升高，弥漫型组织中最高。与 NGFs 相比，CAFs 中 c-Ski 和 α-SMA 的表达均显著升高。当 NGFs 中过表达 c-Ski 时，细胞活力、细胞侵袭和迁移均增强，Smad3 的表达下调。当抑制 c-Ski 时，细胞活力、细胞侵袭和迁移均受到抑制，Smad3 的表达上调。同时，Smad3 的过表达显著逆转了 NGFs 中过表达 c-Ski 的作用，而 Smad3 的敲低则显著逆转了 CAFs 中 si-c-Ski 的作用。过表达 c-Ski 可增强 GC CAFs 的细胞活力，促进细胞侵袭和迁移，其作用可能通过调节 Smad3 信号通路实现。本研究可能为 c-Ski 与 CAFs 之间的关系以及 c-Ski 在癌症发展中的作用提供更深入的认识，并为 GC 的治疗提供新的研究靶点。
