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Functional Validation of DownRegulated MicroRNAs in HeLa Cells Treated with Polyalthia longifolia Leaf Extract Using Different Microscopic Approaches: A Morphological Alteration-Based Validation

Part of: Micrographia Collection

Published online by Cambridge University Press:  06 August 2019

Shanmugapriya ,
Soundararajan Vijayarathna  and
Sreenivasan Sasidharan
Shanmugapriya
Affiliation:
Institute for Research in Molecular Medicine, Universiti Sains Malaysia, 11800 Gelugor, Pulau Pinang, Malaysia
Soundararajan Vijayarathna
Affiliation:
Institute for Research in Molecular Medicine, Universiti Sains Malaysia, 11800 Gelugor, Pulau Pinang, Malaysia
Sreenivasan Sasidharan*
Affiliation:
Institute for Research in Molecular Medicine, Universiti Sains Malaysia, 11800 Gelugor, Pulau Pinang, Malaysia
*
*Author for correspondence: Sreenivasan Sasidharan, E-mail: srisasidharan@yahoo.com
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Abstract

Several microscopy methods have been developed to assess the morphological changes in cells in the investigations of the mode of cell death in response to a stimulus. Our recent finding on the treatment of the IC50 concentration (26.67 μg/mL) of Polyalthia longifolia leaf extract indicated the induction of apoptotic cell death via the regulation of miRNA in HeLa cells. Hence, the current study was conducted to validate the function of these downregulated microRNAs in P. longifolia-treated HeLa cells using microscopic approaches. These include scanning electron microscope (SEM), transmission electron microscope (TEM), and acridine orange/propidium iodide (AO/PI)-based fluorescent microscopy techniques by observing the morphological alterations to cells after transfection with mimic miRNA. Interestingly, the morphological changes observed in this study demonstrated the apoptotic hallmarks, for instance, cell blebbing, cell shrinkage, cytoplasmic and nuclear condensation, vacuolization, cytoplasmic extrusion, and the formation of apoptotic bodies, which proved the role of dysregulated miRNAs in apoptotic HeLa cell death after treatment with the P. longifolia leaf extract. Conclusively, the current study proved the crucial role of downregulated miR-484 and miR-221-5p in the induction of apoptotic cell death in P. longifolia-treated HeLa cells using three approaches—SEM, TEM, and AO/PI-based fluorescent microscope.

Keywords

apoptosisfluorescent microscopeP. longifoliascanning electron microscopytransmission electron microscopy
Type
Micrographia
Information
Microscopy and Microanalysis , Volume 25 , Issue 5 , October 2019 , pp. 1263 - 1272
Copyright
Copyright © Microscopy Society of America 2019 

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