Table_1_Lycorine Induces Mitochondria-Dependent Apoptosis in Hepatoblastoma HepG2 Cells Through ROCK1 Activation.doc

Lycorine, a naturally occurring compound extracted from the Amaryllidaceae plant family, has been reported to exhibit antitumor activity in various cancer cell types. In the present study, we investigated the molecular mechanisms underlying lycorine-induced apoptosis in hepatoblastoma HepG2 cells. We found that lycorine induced mitochondria-dependent apoptosis in HepG2 cells accompanied by mitochondrial permeability transition pore (mPTP) opening, mitochondrial membrane potential (MMP) loss, adenosine triphosphate (ATP) depletion, Ca2+ and cytochrome c (Cyto C) release, as well as caspase activation. Furthermore, we found Rho associated coiled-coil containing protein kinase 1 (ROCK1) cleavage/activation played a critical role in lycorine-induced mitochondrial apoptosis. In addition, the ROCK inhibitor Y-27632 was employed, and we found that co-treatment with Y-27632 attenuated lycorine-induced mitochondrial injury and cell apoptosis. Meanwhile, an in vivo study revealed that lycorine inhibited tumor growth and induced apoptosis in a HepG2 xenograft mouse model in association with ROCK1 activation. Taken together, all these findings suggested that lycorine induced mitochondria-dependent apoptosis through ROCK1 activation in HepG2 cells, and this may be a theoretical basis for lycorine’s anticancer effects.

石蒜碱，一种从石蒜科植物中提取的自然化合物，据报告在各种癌细胞类型中表现出抗肿瘤活性。在本研究中，我们探讨了石蒜碱诱导肝母细胞瘤 HepG2 细胞凋亡的分子机制。我们发现石蒜碱通过诱导线粒体依赖性凋亡在 HepG2 细胞中发挥作用，伴随线粒体通透性转换孔（mPTP）开放、线粒体膜电位（MMP）下降、三磷酸腺苷（ATP）耗竭、钙离子和细胞色素c（Cyto C）释放，以及caspase 激活。此外，我们还发现Rho 相关联的螺旋卷曲蛋白激酶1（ROCK1）的切割/激活在石蒜碱诱导的线粒体凋亡中发挥了关键作用。此外，使用 ROCK 抑制剂 Y-27632 进行了实验，我们发现与 Y-27632 联合处理减弱了石蒜碱诱导的线粒体损伤和细胞凋亡。同时，体内研究表明，石蒜碱通过 ROCK1 激活抑制了 HepG2 异种移植小鼠模型中的肿瘤生长并诱导了细胞凋亡。综合以上发现，所有这些结果表明石蒜碱通过激活 ROCK1 在 HepG2 细胞中诱导线粒体依赖性凋亡，这可能是石蒜碱抗癌作用的理论基础。