分析蛋白质-蛋白质相互作用网络发现巨蹄抑制肺癌转移的作用靶点为 c-Met。
Analysis of the Protein-Protein Interaction Network Identifying c-Met as a Target of Gigantol in the Suppression of Lung Cancer Metastasis.
机构信息
Department of Clinical Pathology, Faculty of Medicine Vajira Hospital, Navamindradhiraj University, Bangkok, Thailand.
Department of Pharmacology and Biopharmaceutics, Faculty of Pharmaceutical Sciences, Huachiew Chalermprakiet University, Samutprakarn, Thailand.
出版信息
Cancer Genomics Proteomics. 2021 May-Jun;18(3):261-272. doi: 10.21873/cgp.20257.
BACKGROUND/AIM: c-Met (mesenchymal-epithelial transition factor) facilitates cancer progression and is recognized as a promising drug target. The molecular target of gigantol from Dendrobium draconis in suppressing cancer metastasis is largely unknown.
MATERIALS AND METHODS
Proteins affected by gigantol treatment were subjected to proteomic and bioinformatic analysis. Protein-Protein interaction (PPI) networks were constructed by the Search Tool for the Retrieval of Interacting Genes (STRING). The Kyoto Encyclopedia of Genes and Genomes (KEGG) database and hub gene were used to enrich the dominant pathways. Western blot analysis and immunofluorescence were used to validate the effect of gigantol on the target protein and signaling.
RESULTS
Gigantol down-regulates 41 adhesion proteins and 39-migratory proteins, while it up-regulates 30 adhesion-related proteins and 22 proteins controlling cell migration. The key components of our constructed PPI network comprised 41 proteins of cell adhesion enriched in 40 nodes with 25 edges, 39 proteins of cell migration enriched in 39 nodes with 76 edges in down-regulated proteins, 30 proteins of cell adhesion enriched in 30 nodes with 21 edges, and 22 proteins of cell migration enriched in 22 nodes with 22 edges in up-regulated protein. c-Met was identified as a central protein of the PPI network in the largest degree. KEGG mapper further suggested that c-Met, PI3K, and AKT were the regulatory proteins affected by gigantol. To confirm, the effects of gigantol on c-Met, the p-PI3K, PI3K, p-AKT, and AKT proteins were investigated by western blotting and the results showed a consistent effect of gigantol in the suppression of the c-Met/PI3K/AKT signal. Next, immunofluorescence showed a dramatic decrease in c-Met, PI3K and AKT activation in response to gigantol.
CONCLUSION
c-Met is an important target of gigantol treatment in lung cancer cells. Gigantol suppresses metastasis-related cell motility through decreasing c-Met resulting in PI3K/AKT signaling disruption.
背景/目的：c-Met（间质上皮转化因子）促进癌症进展，被认为是一个有前途的药物靶点。然而，铁皮石斛中 gigantol 抑制癌症转移的分子靶点在很大程度上尚不清楚。
材料与方法
采用蛋白质组学和生物信息学分析 gigantol 处理后受影响的蛋白质。通过 Search Tool for the Retrieval of Interacting Genes（STRING）构建蛋白质-蛋白质相互作用（PPI）网络。京都基因与基因组百科全书（KEGG）数据库和枢纽基因用于富集优势通路。采用 Western blot 分析和免疫荧光法验证 gigantol 对靶蛋白和信号的影响。
结果
gigantol 下调 41 种黏附蛋白和 39 种迁移蛋白，同时上调 30 种黏附相关蛋白和 22 种控制细胞迁移的蛋白。我们构建的 PPI 网络的关键组成部分包括 41 种细胞黏附蛋白，富集于 40 个节点，有 25 个边；下调蛋白中 39 种细胞迁移蛋白，富集于 39 个节点，有 76 个边；上调蛋白中 30 种细胞黏附蛋白，富集于 30 个节点，有 21 个边；上调蛋白中 22 种细胞迁移蛋白，富集于 22 个节点，有 22 个边。c-Met 被确定为 PPI 网络中程度最大的核心蛋白。KEGG mapper 进一步表明，c-Met、PI3K 和 AKT 是受 gigantol 影响的调节蛋白。为了证实这一点，通过 Western blot 研究了 gigantol 对 c-Met、p-PI3K、PI3K、p-AKT 和 AKT 蛋白的影响，结果显示 gigantol 一致抑制 c-Met/PI3K/AKT 信号。接下来，免疫荧光显示 gigantol 显著降低了 c-Met、PI3K 和 AKT 的激活。
结论
c-Met 是 gigantol 治疗肺癌细胞的重要靶点。gigantol 通过降低 c-Met 抑制与转移相关的细胞迁移，从而破坏 PI3K/AKT 信号。
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