UKM Medical Molecular Biology Institute (UMBI), UKM Medical Center, Jalan Ya'acob Latiff, Bandar Tun Razak, Cheras, 56000, Kuala Lumpur, Malaysia.
Department of Biochemistry, Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Ya'acob Latiff, Bandar Tun Razak, Cheras, 56000, Kuala Lumpur, Malaysia.
J Nat Med. 2019 Sep;73(4):745-760. doi: 10.1007/s11418-019-01323-6. Epub 2019 Jun 8.
Our previous study reported that combined treatment of γ-tocotrienol with 6-gingerol showed promising anticancer effects by synergistically inhibiting proliferation of human colorectal cancer cell lines. This study aimed to identify and elucidate molecular mechanisms involved in the suppression of SW837 colorectal cancer cells modulated by combined treatment of γ-tocotrienol and 6-gingerol. Total RNA from both untreated and treated cells was prepared for transcriptome analysis using RNA sequencing techniques. We performed high-throughput sequencing at approximately 30-60 million coverage on both untreated and 6G + γT3-treated cells. The results showed that cancer-specific differential gene expression occurred and functional enrichment pathway analysis suggested that more than one pathway was modulated in 6G + γT3-treated cells. Combined treatment with 6G + γT3 augmented its chemotherapeutic effect by interfering with the cell cycle process, downregulating the Wnt signalling pathway and inducing apoptosis mainly through caspase-independent programmed cell death through mitochondrial dysfunction, activation of ER-UPR, disruption of DNA repair mechanisms and inactivation of the cell cycle process through the downregulation of main genes in proliferation such as FOXM1 and its downstream genes. The combined treatment exerted its cytotoxic effect through upregulation of genes in stress response activation and cytostatic effects demonstrated by downregulation of main regulator genes in the cell cycle. Selected genes involved in particular pathways including ATF6, DDIT3, GADD34, FOXM1, CDK1 and p21 displayed concordant patterns of gene expression between RNA sequencing and RT-qPCR. This study provides new insights into combined treatment with bioactive compounds not only in terms of its pleiotropic effects that enhance multiple pathways but also specific target genes that could be exploited for therapeutic purposes, especially in suppressing cancer cell growth.
我们之前的研究报告称，γ-生育三烯酚与 6-姜酚联合治疗通过协同抑制人结肠直肠癌细胞系的增殖显示出有前景的抗癌效果。本研究旨在鉴定和阐明由 γ-生育三烯酚和 6-姜酚联合治疗调节的 SW837 结肠直肠癌细胞中涉及的分子机制。未处理和处理过的细胞的总 RNA 均用于通过 RNA 测序技术进行转录组分析。我们对未处理和 6G+γT3 处理的细胞进行了大约 30-6000 万覆盖度的高通量测序。结果表明，发生了癌症特异性差异基因表达，功能富集途径分析表明，6G+γT3 处理的细胞中至少有一条途径被调节。6G+γT3 的联合治疗通过干扰细胞周期过程、下调 Wnt 信号通路和主要通过线粒体功能障碍、内质网未折叠蛋白反应的激活、DNA 修复机制的破坏和通过下调增殖过程中的主要基因（如 FOXM1 及其下游基因）来诱导细胞凋亡，增强其化疗效果。联合治疗通过上调应激反应激活和细胞停滞作用中的基因发挥其细胞毒性作用，这表现为下调细胞周期过程中的主要调节基因。涉及特定途径的选定基因，包括 ATF6、DDIT3、GADD34、FOXM1、CDK1 和 p21，在 RNA 测序和 RT-qPCR 之间显示出基因表达的一致模式。本研究为生物活性化合物的联合治疗提供了新的见解，不仅涉及增强多条途径的多效作用，还涉及可用于治疗目的的特定靶基因，特别是在抑制癌细胞生长方面。
