Discipline of Medical Biochemistry and Chemical Pathology, School of Laboratory Medicine and Medical Sciences, College of Health Sciences, University of KwaZulu-Natal , Durban , South Africa.
Department of Chemistry, Faculty of Applied Sciences, Durban University of Technology , Durban , South Africa.
Nutr Cancer. 2019;71(7):1165-1174. doi: 10.1080/01635581.2019.1597136. Epub 2019 Apr 4.
Hepatocellular carcinoma is one of the leading global epidemics. A medicinal tree, (MO), has been part of traditional treatments including cancer therapies. We investigated the apoptosis inducing effects of MO crude aqueous leaf extract (MOE) in human liver hepatocellular carcinoma (HepG) cells. HepG, PBMCs and Hek293 cell viability was evaluated using MTT assay. Oxidative stress and DNA damage was determined using TBARS and comet assays, respectively. Apoptosis was assessed by caspase-9, -3/7 activities and ATP levels (luminometry). Cell cycle, γH2AX, and cleaved PARP-1 were determined (flow cytometry). Protein expression of c-myc, Bax, p-Bcl2, Smac/DIABLO, Hsp70, SRp30a and cleaved PARP-1 was assessed using western blotting. MOE displayed minimal toxicity in PBMCs and Hek293 cells for 24 h. HepG cells were exposed to MOE (24 h) and an IC (4.479 mg/mL) was determined. MOE significantly increased lipid peroxidation, DNA damage and γH2AX levels. A significant decrease in G1, S and G2-M phase was seen. Significant increase in SRp30a protein expression activated caspase-9. Caspase-9 and -3/7 was significantly increased with significant decrease in ATP levels. Apoptosis was confirmed with significant decrease in c-myc, p-Bcl2 and Hsp70 protein expression and a significant increase in Bax, Smac/DIABLO and PARP-1 cleavage. MOE induces cell-cycle arrest and apoptosis in cancerous HepG cells.
肝细胞癌是全球主要的流行疾病之一。药用树（MO）一直是传统治疗方法的一部分，包括癌症治疗。我们研究了 MO 粗水提叶提取物（MOE）对人肝癌（HepG）细胞的诱导凋亡作用。使用 MTT 法评估 HepG、PBMC 和 Hek293 细胞的活力。分别使用 TBARS 和彗星试验测定氧化应激和 DNA 损伤。通过 caspase-9、-3/7 活性和 ATP 水平（发光法）评估细胞凋亡。通过细胞周期、γH2AX 和裂解的 PARP-1（流式细胞术）来确定。使用 Western blot 检测 c-myc、Bax、p-Bcl2、Smac/DIABLO、Hsp70、SRp30a 和裂解的 PARP-1 的蛋白表达。MOE 在 24 小时内对 PBMC 和 Hek293 细胞显示出最小的毒性。将 HepG 细胞暴露于 MOE（24 小时）并确定 IC（4.479mg/mL）。MOE 显著增加脂质过氧化、DNA 损伤和γH2AX 水平。G1、S 和 G2-M 期明显减少。SRp30a 蛋白表达的显著增加激活了 caspase-9。caspase-9 和 -3/7 显著增加，同时 ATP 水平显著降低。c-myc、p-Bcl2 和 Hsp70 蛋白表达的显著降低和 Bax、Smac/DIABLO 和 PARP-1 切割的显著增加证实了细胞凋亡。MOE 诱导癌细胞 HepG 细胞的细胞周期停滞和凋亡。
