Department of Chemistry, Faculty of Science, Suez Canal University, Ismailia 41522, Egypt.
Department of Chemistry, Faculty of Science, Kafrelsheikh University, Kafr El Sheikh 33516, Egypt.
Molecules. 2021 Dec 23;27(1):83. doi: 10.3390/molecules27010083.
Hepatocellular carcinoma (HCC) is one of the most widespread malignancies and is reported as the fourth most prevalent cause of cancer deaths worldwide. Therefore, we aimed to investigate the probable mechanistic cytotoxic effect of the promising 2-thioxoimidazolidin-4-one derivative on liver cancer cells using in vitro and in vivo approaches. The compounds were tested for the in vitro cytotoxic activity using MTT assay, and the promising compound was tested in colony forming unit assay, flow cytometric analysis, RT-PCR, Western blotting, in vivo using SEC-carcinoma and in silico to highlight the virtual mechanism of action. Both compounds and performed cytotoxic effects against HepG2 cells with IC values of 0.017 and 0.18 μM, respectively, compared to Staurosporine and 5-Fu as reference drugs with IC values of 5.07 and 5.18 µM, respectively. Compound treatment revealed apoptosis induction by 19.35-fold (11.42% compared to 0.59% in control), arresting the cell cycle at G2/M phase. Moreover, studying gene expression that plays critical roles in cell cycle and apoptosis by RT-PCR demonstrated that compound enhances the expression of the pro-apoptotic genes p53, PUMA, and Caspase 3, 8, and 9, and impedes the anti-apoptotic Bcl-2 gene in the HepG2 cells. It can also inhibit the PI3K/AKT pathway at both gene and protein levels, which was reinforced by the in silico predictions of the molecular docking simulations towards the PI3K/AKT proteins. Finally, in vivo study verified that compound has a promising anti-cancer activity through activating antioxidant levels (CAT, SOD and GSH) and ameliorating hematological, biochemical, and histopathological findings.
肝细胞癌（HCC）是最常见的恶性肿瘤之一，据报道是全球癌症死亡的第四大主要原因。因此，我们旨在通过体外和体内方法研究有前途的 2-硫代亚氨唑烷-4-酮衍生物对肝癌细胞的可能机制细胞毒性作用。使用 MTT 测定法测试化合物的体外细胞毒性活性，并用集落形成单位测定法、流式细胞分析、RT-PCR、Western blot 测试有前途的化合物，在 SEC-肝癌体内和计算机模拟中突出虚拟作用机制。与参考药物 Staurosporine 和 5-Fu（IC 值分别为 5.07 和 5.18 µM）相比，化合物 和 对 HepG2 细胞均表现出细胞毒性作用，IC 值分别为 0.017 和 0.18 μM。化合物 处理显示通过 19.35 倍诱导细胞凋亡（与对照相比为 0.59%，增加到 11.42%），将细胞周期阻滞在 G2/M 期。此外，通过 RT-PCR 研究在细胞周期和凋亡中发挥关键作用的基因表达表明，化合物 增强了促凋亡基因 p53、PUMA 和 Caspase 3、8 和 9 的表达，并抑制了 HepG2 细胞中的抗凋亡 Bcl-2 基因。它还可以抑制 PI3K/AKT 通路在基因和蛋白水平，这是通过计算机模拟对 PI3K/AKT 蛋白进行分子对接模拟得到的。最后，体内研究证实化合物 通过激活抗氧化剂水平（CAT、SOD 和 GSH）和改善血液学、生化学和组织病理学发现，具有有前途的抗癌活性。
