Lappaconitine Sulfate Inhibits Proliferation and Induces Apoptosis in Human Hepatocellular Carcinoma HepG2 Cells through the Reactive Oxygen Species-Dependent Mitochondrial Pathway

Abstract

Background: 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. Materials and Methods: 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. Results: 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. Conclusions: Our results demonstrated that LS is a promising therapeutic agent for HCC directed toward the proliferation inhibition and the induction of apoptosis.

C18-diterpenoid alkaloids in tribe Delphineae (Ranunculaceae): phytochemistry, chemotaxonomy, and bioactivities

“…It was reported that the hydrobromide salt of lappaconitine could suppress the growth of liver and S180 tumors of mice with inhibition rates of 11.20%∼53.08% and 29.81%∼53.96%, respectively 111,112 and could inhibit the proliferation of human non-small cell lung cancer A549 cells dose dependently by arresting the cells in G1/G0 phase and downregulating the expression of Cyclin E1 ( Table 1 ). 113 Lappaconitine sulfate was also reported to possess antiproliferative activity against various human cancer cell lines: cerical neoplasm (HeLa), 114 liver (HepG2), 115 colon (HT-29), 116 and lung (A549), 117 which might be caused by activation of p38 MAPK-, mitochondrial-, and caspase-mediated apoptosis. In addition, the hydrochloride salt of lappaconitine has been observed for its ability to inhibit proliferation and to induce apoptosis in human colon cancer HCT-116 cells and human liver cancer HepG2 cells via mitochondrial and MAPK pathways.…”

Synthesis, Pharmacological Evaluation, and Molecular Modeling of Lappaconitine–1,5-Benzodiazepine Hybrids

“…Lappaconitine sulfate [ 29 ], lappaconitine hydrobromide, and lappaconitine trifluoroacetate [ 30 ] have exhibited greater solubility in water and pronounced analgesic activity. Salt formation with sulfonic acid was important for antitumor activity [ 31 , 32 ]. A fatty acid chain at the 8-hydroxy group and 13-OH substituent on the diterpenoid core were favorable for the enhancement of the antiproliferative activity [ 33 ].…”

Frontier progress of the combination of modern medicine and traditional Chinese medicine in the treatment of hepatocellular carcinoma

“…Alperine inhibited Akt-mediated apoptosis, G2/M cell cycle arrest and the proliferation of hepatoma cells [ 28 ]. Lappaconitine sulfate induced apoptosis by mediating the mitochondrial apoptosis pathway [ 29 ]. Bufalin regulated tumor immune microenvironments by Nuclear Factor kappa B (NF-κB) and activates anti-tumor T cell immune response [ 30 ].…”