The Novel Herbal Cocktail AGA Alleviates Oral Cancer through Inducing Apoptosis, Inhibited Migration and Promotion of Cell Cycle Arrest at SubG1 Phase

SIMPLE SUMMARY: Oral cancer is a major cause of death from oral disorders worldwide. These are highly lethal, incapacitating and present mostly in the form of squamous cell carcinomas. Owing to therapy resistance, the impact of conventional drugs is limited. Therefore, we aimed to develop a novel anti-oral cancer therapeutic alternative in the form of our synthesized robust cocktail of Antler’s extract (A) and Ganoderma lucidum (G) and Antrodia Camphorata (A), designated as AGA. We demonstrated that AGA, primarily contained with bioactive components such as triterpenoids and polysaccharides strongly, induces cellular apoptosis and inhibits cell migration and cyclin expression, leading to promoted cell cycle arrest at the subG1 phase. AGA could also efficaciously retard tumor growth without any toxic influence on liver and kidneys. Therefore, AGA holds a great potential to be an oral cancer treatment strategy. ABSTRACT: Traditional Chinese medicines Antler’s extract (A) and Ganoderma lucidum (G) and Antrodia Camphorata (A) have been known to individually contain a plethora of bioactive factors including triterpenoids, polysaccharides etc., exerting various curative impacts such as anti-inflammatory, anti-oxidative, anti-atherosclerotic and anti-viral activities. However, their combinatorial therapeutic efficacy for oral cancer has not been investigated. Hence, we synthesized a robust cocktail called AGA and investigated its anti-oral cancer potential in vitro and in vivo. An MTT assay revealed the IC(50) of AGA to be about 15 mg at 72 h. Therefore, 10 mg and 20 mg doses were selected to study the effect of AGA. The AGA significantly inhibited proliferation of oral cancer cells (HSC3, SAS, and OECM-1) in a dose- and time-dependent manner. AGA retarded cell cycle regulators (CDK4, CDK6, cyclin A, B1, D1 and E2) and apoptosis inhibitory protein Bcl-2, but enhanced pro-apoptotic protein Bax and a higher percentage of cells in Sub-G1 phase. Mechanistically, AGA suppressed all EMT markers; consequently, it decreased the migration ability of cancer cells. AGA significantly reduced xenograft tumor growth in nude mice with no adverse events in liver and renal toxicity. Conclusively, AGA strongly inhibited oral cancer through inducing apoptosis and inhibiting the migration and promotion of cell cycle arrest at subG1 phase, which may be mediated primarily via cocktail-contained triterpenoids and polysaccharides.