Zoology Graduate Program, Department of Zoology, Faculty of Science, Cairo University, Giza 12613, Egypt.
Biochemistry and Molecular Biology Unit, Department of Cancer Biology, National Cancer Institute, Cairo University, Giza, 12613, Egypt.
Asian Pac J Cancer Prev. 2024 Oct 1;25(10):3725-3739. doi: 10.31557/APJCP.2024.25.10.3725.
This study investigated the potential of m-TOR inhibitor drugs (sirolimus, everolimus, and tacrolimus) in combating both hepatocellular carcinoma (HCC) and hepatitis C virus (HCV) replication.
After treating HepG2 and PBMCs with the mammalian target of Rapamycin (m-TOR) inhibitors drugs; sirolimus, everolimus, and tacrolimus at different concentrations (1, 5, and 10 µM/µl), cell proliferation was assessed using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Antioxidant activities (total antioxidant, glutathione S-transferase, and glutathione reductase), Fas-ligand level, tumor necrosis factor-α (TNF-α) level, caspase-3, -8, and -9 activities, and cell cycle analysis were measured. quantitative Real-time PCR, colony forming assay, molecular docking studies after infection of PBMCs with 1 ml (1.5 × 106 HCV) serum then incubated with m-TOR inhibitor drugs at their respective IC50 concentrations.
In HepG2 cells, treatment with these inhibitors resulted in suppressed cell viability, increased dead cell accumulation, and enhanced apoptotic signaling through elevated Fas-ligand and caspase activities. Additionally, cell cycle analysis revealed arrest in G0/G1 and G2/M phases, further hindering HCC progression. Furthermore, m-TOR inhibitor drugs significantly reduced HCV viral load and colony formation in infected PBMCs. This antiviral effect was accompanied by decreased TNF-α activity, suggesting potential modulation of the inflammatory response associated with HCV infection. Molecular docking studies provided theoretical support for these findings, with Sovaldi demonstrating the highest binding affinity towards key HCV targets compared to other m-TOR inhibitors. This suggests its potential as a potent HCV inhibitor, while also highlighting the potential of exploring m-TOR inhibitors for future HCV treatment development.
Overall, this study provides encouraging evidence for the potential of m-TOR inhibitor drugs as promising therapeutic agents for both HCC and HCV, warranting further investigation and optimization for clinical applications.
本研究旨在探讨哺乳动物雷帕霉素靶蛋白（mTOR）抑制剂药物（西罗莫司、依维莫司和他克莫司）在抑制肝细胞癌（HCC）和丙型肝炎病毒（HCV）复制方面的潜力。
用不同浓度（1、5 和 10 μM/μl）的 mTOR 抑制剂药物西罗莫司、依维莫司和他克莫司处理 HepG2 和 PBMCs 后，通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法评估细胞增殖。测量抗氧化活性（总抗氧化、谷胱甘肽 S-转移酶和谷胱甘肽还原酶）、Fas 配体水平、肿瘤坏死因子-α（TNF-α）水平、caspase-3、-8 和 -9 活性以及细胞周期分析。在感染 PBMCs 后，用 1ml（1.5×106 HCV）血清孵育 mTOR 抑制剂药物，然后在各自的 IC50 浓度下进行定量实时 PCR、集落形成测定和分子对接研究。
在 HepG2 细胞中，这些抑制剂的治疗导致细胞活力受到抑制，死亡细胞积累增加，通过 Fas 配体和半胱天冬酶活性的升高增强了凋亡信号。此外，细胞周期分析显示 G0/G1 和 G2/M 期停滞，进一步阻止 HCC 进展。此外，mTOR 抑制剂药物显著降低了感染 PBMCs 中的 HCV 病毒载量和集落形成。这种抗病毒作用伴随着 TNF-α 活性的降低，表明可能调节与 HCV 感染相关的炎症反应。分子对接研究为这些发现提供了理论支持，Sovaldi 与其他 mTOR 抑制剂相比，对关键 HCV 靶标表现出最高的结合亲和力。这表明它有作为一种有效的 HCV 抑制剂的潜力，同时也强调了探索 mTOR 抑制剂用于未来 HCV 治疗开发的潜力。
总的来说，这项研究为 mTOR 抑制剂药物作为 HCC 和 HCV 有希望的治疗药物提供了令人鼓舞的证据，值得进一步研究和优化以用于临床应用。
