Department of Regenerative Medicine, School of Pharmaceutical Science, Jilin University, Fujin Road 1266, Changchun 130021, China.
Department of Ophthalmology, The First Hospital of Jilin University, Jilin University, Xinmin Street 1, Changchun 130021, China.
Molecules. 2022 Aug 6;27(15):5002. doi: 10.3390/molecules27155002.
The mechanism of ginsenoside Rh3 activity against cancer remains unclear. This study aimed to investigate the underlying mechanism. The effects of Rh3 on the cell proliferation, migration and invasion, and cycle and apoptosis were analyzed using CCK-8 assay, transwell migration assay and flow cytometry, respectively. The RNA transcriptome was sequenced and data were analyzed by R software. Protein expression and protein-protein interactions were determined by Western blotting and co-immunoprecipitation, respectively. The results showed Rh3 inhibited HCT116 cell proliferation, invasion, and migration, arrested cells at G1 phase; and increased apoptosis. Rh3 downregulated 314 genes and upregulated 371 genes. Gene Set Enrichment Analysis (GSEA) using ranked DNA replication first, while GSEA using Gene Ontology ranked the initiation of DNA replication first. Compared with tumor data from The Cancer Genome Atlas (TCGA), most of genes related to DNA replication were oppositely regulated by Rh3. Furthermore, Rh3 down-regulated key protein expression related to DNA replication (Orc6, Cdt1, and Mcm2), but did not affect the loading of Mcm complexes onto ORC complexes nor the phosphorylation at ser139 of Mcm2. Therefore, Rh3 may inhibit colorectal cancer HCT116 cells by downregulation of genes related to DNA replication.
人参皂苷 Rh3 抑制结直肠癌细胞增殖、迁移和侵袭的作用机制及其与 DNA 复制的关系
人参皂苷 Rh3 抑制肿瘤的作用机制尚不清楚。本研究旨在探讨其作用机制。采用 CCK-8 法、Transwell 迁移实验和流式细胞术分别分析 Rh3 对细胞增殖、迁移和侵袭以及细胞周期和凋亡的影响。对 RNA 转录组进行测序，并用 R 软件进行数据分析。通过 Western blot 和免疫共沉淀分别检测蛋白表达和蛋白-蛋白相互作用。结果表明，Rh3 抑制 HCT116 细胞增殖、侵袭和迁移，将细胞阻滞在 G1 期，并诱导细胞凋亡。Rh3 下调 314 个基因，上调 371 个基因。使用排名 DNA 复制的基因集富集分析（GSEA），结果显示 DNA 复制排名第一，而使用基因本体论（GO）排名 DNA 复制起始排名第一。与癌症基因组图谱（TCGA）中的肿瘤数据相比，大多数与 DNA 复制相关的基因被 Rh3 相反调控。此外，Rh3 下调与 DNA 复制相关的关键蛋白表达（Orc6、Cdt1 和 Mcm2），但不影响 Mcm 复合物在 ORC 复合物上的加载，也不影响 Mcm2 丝氨酸 139 的磷酸化。因此，Rh3 可能通过下调与 DNA 复制相关的基因来抑制结直肠癌细胞 HCT116 的增殖、迁移和侵袭。
