Department of Drug Discovery & Biomedical Sciences, Medical University of South Carolina, Charleston, SC, USA; Institute for Immunological and Physiopathological Studies (IIFP), National Scientific and Technical Research Council (CONICET), Argentina.
Department of Drug Discovery & Biomedical Sciences, Medical University of South Carolina, Charleston, SC, USA.
Eur J Pharmacol. 2023 Oct 15;957:176034. doi: 10.1016/j.ejphar.2023.176034. Epub 2023 Aug 29.
Voltage dependent anion channels (VDAC) in the outer mitochondrial membrane regulate the influx of metabolites that sustain mitochondrial metabolism and the efflux of ATP to the cytosol. Free tubulin and NADH close VDAC. The VDAC-binding small molecules X1 and SC18 modulate mitochondrial metabolism. X1 antagonizes the inhibitory effect of tubulin on VDAC. SC18 occupies an NADH-binding pocket in the inner wall of all VDAC isoforms. Here, we hypothesized that X1 and SC18 have a synergistic effect with sorafenib, regorafenib or lenvatinib to arrest proliferation and induce death in hepatocarcinoma cells. We used colony formation assays to determine cell proliferation, and a combination of calcein/propidium iodide, and trypan blue exclusion to assess cell death in the well differentiated Huh7 and the poorly differentiated SNU-449 cells. Synergism was assessed using the Chou-Talalay method. The inhibitory effect of X1, SC18, sorafenib, regorafenib and lenvatinib was concentration and time dependent. IC calculated from the inhibition of clonogenic capacity were lower than those determined from cell survival. At IC that inhibited cell proliferation, SC18 arrested cells in G0/G1. SC18 at 0.25-2 IC had a synergistic effect with sorafenib on clonogenic inhibition in Huh7 and SNU-449 cells, and with regorafenib or lenvatinib in SNU-449 cells. X1 or SC18 also had synergistic effects with sorafenib on promoting cell death at 0.5-2 IC for SC18 in Huh7 and SNU-449 cells. These results suggest that small molecules targeting VDAC represent a potential new class of drugs to treat liver cancer.
电压依赖性阴离子通道 (VDAC) 位于线粒体外膜，调节维持线粒体代谢所需的代谢物的流入和 ATP 向细胞质的流出。游离微管蛋白和 NADH 使 VDAC 关闭。VDAC 结合的小分子 X1 和 SC18 调节线粒体代谢。X1 拮抗微管蛋白对 VDAC 的抑制作用。SC18 占据所有 VDAC 同工型内壁的 NADH 结合口袋。在这里，我们假设 X1 和 SC18 与索拉非尼、瑞戈非尼或仑伐替尼具有协同作用，可阻止肝癌细胞增殖并诱导其死亡。我们使用集落形成测定法来确定细胞增殖，并用钙黄绿素/碘化丙啶和台盼蓝排除法组合评估分化良好的 Huh7 和分化较差的 SNU-449 细胞的细胞死亡。使用 Chou-Talalay 方法评估协同作用。X1、SC18、索拉非尼、瑞戈非尼和仑伐替尼的抑制作用具有浓度和时间依赖性。从抑制集落形成能力计算出的 IC 低于从细胞存活中确定的 IC。在抑制细胞增殖的 IC 下，SC18 将细胞阻滞在 G0/G1 期。SC18 在 0.25-2 IC 时与索拉非尼在 Huh7 和 SNU-449 细胞中的集落抑制具有协同作用，与瑞戈非尼或仑伐替尼在 SNU-449 细胞中具有协同作用。在 0.5-2 IC 时，X1 或 SC18 也与索拉非尼在促进 Huh7 和 SNU-449 细胞中的细胞死亡方面具有协同作用。这些结果表明，靶向 VDAC 的小分子代表了治疗肝癌的一类新的潜在药物。
