Shanmugapriya, Shanmugapriya
(2019)
Molecular Characterization And Bioinformatics Analysis Of Microrna-221-5p Regulated By Standardized Polyalthia Longifolia (Sonn.) Thwaites Leaf Extract In Hela Cell Lines.
PhD thesis, Universiti Sains Malaysia.
Abstract
Polyalthia longifolia (Sonn.) Thwaites is an exquisite plant species with rich ethnomedicinal values. Recent scientific investigations on P. longifolia leaf extract have also revealed its anti-cancer property against HeLa cells through the induction of caspase-dependent apoptosis by regulating microRNA (miRNA) expressions. However, there were no further investigations performed to report the functional analysis of the regulated miRNA, with absolutely no means of scientific evidence of validation of miRNA dysregulation in HeLa cells treated with the methanolic P. longifolia leaf extract. Hence, this study was conducted to validate the miRNA expression in methanolic P. longifolia leaf extract treated HeLa cells in comparison with untreated HeLa cells with an intricate elucidation of functional and proteomic analysis of miRNA expression. In this study, methanolic P. longifolia leaf extract was freshly prepared and MTT assay was performed to identify the IC50 value against HeLa cells. The expression of miR-221-5p was validated by performing Taqman real time RTqPCR which confirmed the down-regulation of miR-221-5p in HeLa cells treated with methanolic P. longifolia leaf extract compared to the untreated HeLa cells. The functional analysis of miR-221-5p was conducted through gain-of-function and loss-of-function approach by MTT assay, flow cytometric analysis of Annexin V/ Propidium Iodide assay and caspase-3 assay.
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