细胞外线粒体膜的电压依赖性孔道（VDAC）调节维持线粒体代谢的代谢物输入和ATP向细胞质的流出。游离的微管蛋白和NADH会关闭VDAC。小分子化合物X1和SC18会打开VDAC，并调节线粒体代谢。X1对抗微管蛋白对VDAC的抑制作用。SC18占据所有VDAC亚型内壁中的NADH结合口袋。在这里，我们假设X1和SC18与索拉非尼、雷格拉非尼或来那替尼具有协同作用，能够抑制肝癌细胞的增殖并诱导细胞死亡。我们使用集落形成实验来确定细胞增殖情况，并结合卡尔色素/丙碘胺和尝试蓝排除法评估细胞死亡情况，其中包括分化良好的Huh7和分化较差的SNU-449细胞。我们使用Chou-Talalay方法评估协同作用。X1、SC18、索拉非尼、雷格拉非尼和来那替尼的抑制作用与浓度和时间有关。通过对克隆形成能力的抑制，IC50值低于细胞存活率计算得出的值。在抑制细胞增殖的IC50浓度下，SC18将细胞停滞在G0/G1期。在Huh7和SNU-449细胞中，SC18在0.25-2倍IC50浓度下与索拉非尼具有协同作用，抑制克隆形成能力，并且在SNU-449细胞中与雷格拉非尼或来那替尼也具有协同作用。在Huh7和SNU-449细胞中，X1或SC18与索拉非尼在0.5-2倍SC18的IC50浓度下也具有促进细胞死亡的协同作用。这些结果表明，靶向VDAC的小分子化合物可能成为治疗肝癌的新型药物。版权所有 © 2023 Elsevier B.V. 发表。

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 small molecules X1 and SC18 open VDAC and 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. IC50 values calculated from the inhibition of clonogenic capacity were lower than those determined from cell survival. At IC50s that inhibited cell proliferation, SC18 arrested cells in G0/G1. SC18 at 0.25-2 IC50s had a synergistic effect with sorafenib on inhibiting clonogenicity 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 IC50s 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.Copyright © 2023. Published by Elsevier B.V.