Cytotoxic and antimigratory effects of Cratoxy formosum extract against HepG2 liver cancer cells

Abstract

Abstract. The aim of the present study was to investigate the molecular mechanisms underlying Cratoxylum formosum (CF) Dyer-induced cancer cell death and antimigratory effects in HepG2 liver cancer cells. The cytotoxic, antiproliferative and antimigratory effects of CF leaf extract on human liver cancer HepG2 cell lines were evaluated using sulforhodamine B, colony formation, and wound healing assays. In addition, apoptosis induction mechanisms were investigated via reactive oxygen species (ROS) formation, caspase 3 activities, and mitochondrial membrane potential (ΔΨm) disruption. Gene expression and apoptosis-associated protein levels were measured by reverse transcription-quantitative polymerase chain reaction and western blotting. CF induced HepG2 cell death in a time-and dose-dependent manner with half maximal inhibitory concentration values of 219.03±9.96 and 124.90±6.86 µg/ml at 24 and 48 h, respectively. Treatment with CF caused a significant and dose-dependent decrease in colony forming ability and cell migration. Furthermore, the present study demonstrated that CF induced ROS formation, increased caspase 3 activities, decreased the ΔΨm, and caused HepG2 apoptosis. CF marginally decreased the expression level of the cell cycle regulatory protein, ras-related C3 botulinum toxin substrate 1 (rho family, small GTP binding protein Rac1) and the downstream protein, cyclin dependent kinase 6. Additionally, CF significantly enhanced p21 levels, reduced cyclin D1 protein levels and triggered cancer cell death. CF leaf extracts induced cell death, stimulated apoptosis and inhibited migration in HepG2 cells. Thus, CF may be useful for developing an anticancer drug candidate for the treatment of liver cancer. IntroductionMedicinal plants and their active compounds have been reported to exert potent cytotoxic activity against various types of cancer cell (1). Numerous plants are consumed as food and are claimed by practitioners of traditional medicine to promote health (2-4). Northeast Thai vegetables, such as Cratoxylum formosum (CF) Dyer may have an affect on human health by exerting antioxidant and anticancer effects. CF belongs to the Guttiferae family and is a plant of the tropics, cultivated in many Southeast Asian countries, including Thailand (5). It is a local dietary and herbal plant, and its leaves are usually consumed fresh. CF has been adopted in folk medicine for the treatment of fever, coughs, stomach ache and peptic ulcers (6,7). CF produces various secondary metabolites, including phenolic compounds, triterpenoids, flavonoids (6,8), xanthones, anthraquinones (7), chlorogenic acid, dicaffeoylquinic acids, and ferulic acid (5). Kukongviriyapan et al (9) detected potent antioxidant activity in aqueous extracts of CF leaves. Other bioactivities demonstrated by CF include anti-inflammatory (10), antibacterial (11), antimicrobial (12) and anticancer (13). Various parts of the plant have been found to exert anticancer effects, including the roots, bark and leaves (14-16).CF inhibits the proliferation of v...

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“…In fact, dietary plants are encompassed in terms of medical values that can serve as invaluable source for anticancer drug development. Moreover, the anti-cancer drugs available in the dietary supplement are quite safe and effective regulation of normal cell metabolism and also accelerating the programmed cell death by interrupting the uncontrolled cell proliferation [28]. It is well-know that the brown seaweeds are being used as medicinal dietary supplements in most of the Asian countries.…”

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“…SRB was used to determine cytotoxicity as previously described (Buranrat et al, 2017). After 24-h exposure, cells were fixed with 10% ice-cold trichloroacetic acid (TCA) and then incubated in darkness at 4°C for an hour.…”