Investigation of polyamines as chemopreventive agent in selected prophetic food plants in human colon cancer-derived metastatic sw620 cells

Abdul Ghani, Radiah and Ibrahim, Muhammad and Jamil, Elyna Fatini (2021) Investigation of polyamines as chemopreventive agent in selected prophetic food plants in human colon cancer-derived metastatic sw620 cells. Technical Report. UNSPECIFIED. (Unpublished)

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Abstract

Introduction: Polyamines are vital in maintaining human health because they performed certain functions that are necessary for cell development. It can be obtained by endogenous synthesis or exogenous supply through diet. Increasing intracellular polyamine content however promotes unwarranted cell proliferation and might as well stimulate cancer initiation. Thus, it is crucial to evaluate polyamine content in herbs that claimed to cure cancer to avoid unnecessary cell growth and proliferation. Therefore, this study aimed to investigate polyamines as chemopreventive agent using selected prophetic medicinal plants on human lung adenocarcinoma cells, A549. Methods: Five prophetic fruits were selected included ajwa dates (Phoenix dactylifera), beetroot (Beta vulgaris), fig (Ficus auriculata), jujube (Ziziphus jujube) and raisin (Vitis vinifera). These fruits were freeze dried with freeze-drying machine and stored in -80°C. Polyamine concentration in each fruit was quantified by High Performance Liquid Chromatography (HPLC). Subsequently, anti-proliferative effect of the fruits was evaluated in A549 cells using MTT assay and IC50 was determined. Later, growth effect of A549 cells was analyzed via Trypan blue exclusion assay and cell number and cell viability were recorded. Cells fraction from growth effect study was then used to measure protein content and intracellular polyamine concentration. From this, correlation between cell number and protein content was obtained. Cell cycle profile and apoptosis assay were analyzed with flow cytometer while caspase assay was done using microtiter plate reader. Apoptosis and caspase assay allowed type and mechanism of cell death to be detemined. Finally, expression of ornithine decarboxylase (ODC) and spermine/spermidine (N1)-acetyltransferase SSAT gene that regulate polyamine metabolic pathway was elucidated using qPCR. Results: Polyamine concentration in selected fruits showed significant highest polyamine concentration in jujube (219.6 ±4.4 nmoles/ml) while lowest in fig (39.3±3.0 nmoles/ml). MTT assay suggested IC50 was ranged from 15 mg/ml to 30 mg/ml. Growth effect analysis indicated significant decreased in cell number and cell viability after 24 hour of treatments. Protein content was decreased significantly and total elimination of intracellular spermidine and spermine was observed in treated A549 cells. There was correlation between cell number and protein content. Cell cycle profile displayed cell cycle arrest at G0/G1 after 48 hour of treatment in all treated cells. Annexin-v results demonstrated significant increase in early apoptosis in beetroot treated cells while other plants caused significant increase in late apoptosis at 48 hour of treatments. Caspase assay revealed significant activation of caspase 8 in cells treated with jujube and raisins while both caspase 8 and caspase 9 were activated in cells treated with ajwa dates, beetroot and fig. Expression of ODC gene and SSAT gene indicated significant downregulation of ODC gene in cells treated with ajwa and fig while upregulation of SSAT gene in beetroot, jujube and raisins treated cells. Conclusion: It was concluded that novel designs of nutritional therapeutic and preventive approaches for cancer using selected prophetic medicinal fruits are promising.

Item Type:	Monograph (Technical Report)
Additional Information:	4458/95193 - MFI 301221
Uncontrolled Keywords:	polyamines, Prophetic food, cancer, chemoprevention, cell death
Subjects:	Q Science > QP Physiology R Medicine > RB Pathology
Kulliyyahs/Centres/Divisions/Institutes (Can select more than one option. Press CONTROL button):	Kulliyyah of Allied Health Sciences > Department of Biomedical Science (Effective:1st July 2011) Kulliyyah of Allied Health Sciences > Department of Nutrition Sciences
Depositing User:	Dr. Radiah Abdul Ghani
Date Deposited:	30 Dec 2021 16:41
Last Modified:	30 Dec 2021 16:41
URI:	http://irep.iium.edu.my/id/eprint/95193

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