College of Life Science, Northwest Normal University, Lanzhou, China.
College of Life Science, Northwest Normal University, Lanzhou, China,
Pharmacology. 2020;105(11-12):705-714. doi: 10.1159/000506081. Epub 2020 Feb 14.
Hepatocellular carcinoma (HCC) is the third leading cause of tumor-related deaths in the word. Lappaconitine (LA), a diterpenoid alkaloid, exerts antitumor activities. However, the effects and mechanisms of LA sulfate (LS) on HCC remain unclear. This study evaluated the activities and explored the underlying mechanisms of LS in HCC cell line HepG2 cells.
The cell viability and proliferation were evaluated using the Cell Counting Kit-8 (CCK-8) and 5-ethynyl-2'-deoxyuridine (EdU) assay, respectively. The cell cycle distribution was detected by propidium iodide (PI) staining assay. The apoptosis was detected by Annexin -V-fluorescein isothiocyanate (FITC)/PI double staining assay. The cell cycle arrest and apoptosis-related proteins were estimated by western blot analysis. The mitochondrial membrane potential (MMP) was -determined through the 5, 5', 6, 6'-tetrachloro-1, 1', 3, 3'-tetraethylbenzimi-dazolyl carbocyanine iodide (JC-1) staining assay. The reactive oxygen species (ROS) was monitored by 20-70-dichlorofluorescein diacetate (DCFH-DA) staining assay. In vivo antitumor activities were investigated by HepG2 xenograft model.
Our results showed that LS significantly -inhibited the viability and proliferation of HepG2 cells. LS triggered G0/G1 cell cycle arrest, apoptosis and caspase activation. Furthermore, LS induced MMP loss and ROS accumulation. Additionally, LS suppressed the phosphatidylinositol-3-kinase (PI3K)/protein kinase B (AKT)/glycogen synthase kinase 3β (GSK3β) signaling pathway. An in vivo assay showed that LS exhibited a pronounced antitumor effect in nude mice bearing HepG2 xenografts.
Our results demonstrated that LS is a promising therapeutic agent for HCC directed -toward the proliferation inhibition and the induction of apoptosis.
肝细胞癌（HCC）是全球肿瘤相关死亡的第三大原因。拉巴科因（LA），一种二萜生物碱，具有抗肿瘤活性。然而，硫酸拉巴科因（LS）对 HCC 的作用和机制尚不清楚。本研究评估了 LS 在 HepG2 细胞系中的活性并探讨了其潜在机制。
使用细胞计数试剂盒-8（CCK-8）和 5-乙炔基-2'-脱氧尿苷（EdU）测定法分别评估细胞活力和增殖。通过碘化丙啶（PI）染色法检测细胞周期分布。通过 Annexin-V-荧光素异硫氰酸酯（FITC）/PI 双染色法检测细胞凋亡。通过 Western blot 分析评估细胞周期阻滞和凋亡相关蛋白。通过 5,5',6,6'-四氯-1,1',3,3'-四乙基苯并咪唑基碳氰化碘（JC-1）染色法测定线粒体膜电位（MMP）。通过 20-70-二氯荧光素二乙酸酯（DCFH-DA）染色法监测活性氧（ROS）。通过 HepG2 异种移植模型研究体内抗肿瘤活性。
我们的结果表明，LS 显著抑制 HepG2 细胞的活力和增殖。LS 引发 G0/G1 细胞周期阻滞、凋亡和 caspase 激活。此外，LS 诱导 MMP 丧失和 ROS 积累。此外，LS 抑制了磷脂酰肌醇-3-激酶（PI3K）/蛋白激酶 B（AKT）/糖原合成酶激酶 3β（GSK3β）信号通路。体内实验表明，LS 在荷瘤裸鼠中表现出明显的抗肿瘤作用。
我们的研究结果表明，LS 是一种有前途的 HCC 治疗药物，可抑制增殖并诱导细胞凋亡。
