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Title:	Mechanistic study on the anti-proliferative effect of herbal formulation C168 on human colorectal carcinoma cells
Authors:	Leong Lek Mun (P64519)
Supervisor:	Nor Fadilah Rajab, Associate Professor Dr.
Keywords:	Herbs Anti-proliferative effect Colorectal carcinoma cells Herbal formulation Dissertations, Academic -- Malaysia
Issue Date:	30-Jul-2017
Description:	The use of herbal formulation has gained interest due to its promising therapeutic properties especially in cancer treatment. The objectives of this study are to investigate the anti-proliferative effect of a herbal formulation denoted as C168 and its underlying mechanism. This herbal formulation is a mixture of eight plant species, namely Cinnamomum spp., Zingiber spp., Atractylodes spp., Carthamus spp., Angelica spp., Curcuma spp., Glycyrrhiza spp., and Astragalus spp. The study is divided into three phase: chemical composition of C168 methanol extract (CME, Phase I), elucidation of anti-proliferative mechanism of CME (Phase II), and molecular targets of CME (Phase III). With the aid of liquid chromatography - mass spectrometer (LC-MS) coupled with dereplication strategy, twenty nine known secondary metabolites were identified in C168 methanol extract (CME) and some have proven anti-proliferative effect. CME exerted anti-proliferative activity toward HCT 116 human colorectal carcinoma cells and HepG2 hepatocellular carcinoma cells but not to CCD-841-CoN human normal colon epithelial cell, CCD-18Co human normal colon fibroblast cells, Jurkat E6.1 lymphoblastic leukemic cells, and V79-4 Chinese hamster lung fibroblast cells, demonstrating its selectivity towards colon cancer cells. Due to the susceptibility of HCT 116 cells to CME as compared to other cell lines, further investigations were conducted on HCT 116 cells as targeted cells. Further investigation revealed that CME induced G2/M cell cycle arrest and apoptosis as the main anti-proliferative event in HCT 116 cells. In order to further understand the molecular events in CME-induced apoptosis, HCT 116 cells were selected as the targeted cells owing to its susceptibility to CME. When caspases were studied across a series of time points, pro-caspase 8 processing was detected, followed by pro-caspase 3 processing that chronologically led to apoptosis. Investigation carried out using specific caspase inhibitor confirmed the role of caspase 8 as an initiator caspase in CME-induced apoptosis. Treatment of CME in HCT 116 cells showed an early induction of oxidative stress via increasing superoxide anion and hydrogen peroxide level as well as decreasing intracellular glutathione level. However, antioxidant Nacetyl cysteine and trolox failed to ameliorate CME-induced apoptosis, suggesting that oxidative stress is not the critical event that led to CME-induced apoptosis. DNA damage assessment revealed the increase in tail moment value and upregulation of phosphorylated H2AX level in CME-treated cells, suggesting DNA damage as an early signal of CME induced-apoptosis. Mitochondria membrane potential (MMP) loss in CME-treated cells was also involved in CME-induced apoptosis. In conclusion, this study highlighted the selectivity of CME-induced anti-proliferative effects on colon cancer cells which involved caspase 8 activation, DNA damage, and mitochondrial membrane potential loss as early events in CME-induced apoptosis.,Ijazah Doktor Falsafah
Pages:	228
Call Number:	QV766.L583m 2017 9
Publisher:	UKM, Kuala Lumpur
Appears in Collections:	Faculty of Health Sciences / Fakulti Sains Kesihatan

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