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Expression of beta defensin genes in frozen thawed and cultured immortalised human corneal epithelial cell line

Human beta defensins (hBD) are important host defence molecules at the ocular surface. In addition to their antimicrobial activities, hBD may also act as regulatory factors in recruiting and activating immune cells. Only hBD-1 – hBD-4 have been well characterised. To date, a complete profile of t...

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Main Authors: Rohayu Izanwati, Mohd Rawi, Nurul Shakirah, Azmi, Yow, Weng Kit, Nurul Hana, Zainal Baharin
Format: Article
Language:English
Published: 2017
Subjects:
R Medicine (General)
Online Access:http://eprints.unisza.edu.my/6049/1/FH02-FSK-19-36067.pdf
http://eprints.unisza.edu.my/6049/
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spelling my-unisza-ir.60492022-05-22T03:54:50Z http://eprints.unisza.edu.my/6049/ Expression of beta defensin genes in frozen thawed and cultured immortalised human corneal epithelial cell line Rohayu Izanwati, Mohd Rawi Nurul Shakirah, Azmi Yow, Weng Kit Nurul Hana, Zainal Baharin R Medicine (General) Human beta defensins (hBD) are important host defence molecules at the ocular surface. In addition to their antimicrobial activities, hBD may also act as regulatory factors in recruiting and activating immune cells. Only hBD-1 – hBD-4 have been well characterised. To date, a complete profile of the beta defensin genes (DEFB) expression in immortalised human corneal epithelial cell line (HCE-2) has not been established. Therefore, this study is aimed to explore a spectrum of DEFB expression in HCE-2. Total RNAs were extracted from frozen thawed HCE-2 and cultured HCE-2 to ensure the gene expression were identical. The RNAs were reverse transcribed into cDNAs. The expression of 10 DEFB (DEFB1, DEFB4A, DEFB103, DEFB104, DEFB105, DEFB106, DEFB109, DEFB123, DEFB126 and DEFB127) were analysed using polymerase chain reaction (PCR) and gel electrophoresis. DEFB1 and DEFB103 were the only hBD mRNAs found constitutively expressed in frozen thawed HCE-2 and cultured HCE-2. It is also interesting to note that PCR enhancer was needed to amplify the genes in cultured HCE-2. Our findings suggest that corneal epithelium constantly produce hBD-1 and hBD-3, which presumably provide the baseline defence against infection. Further investigation on the expression of these genes when HCE-2 stimulated with proinflammatory cytokines would help in better understanding of the ocular surface defence mechanism. 2017-05 Article PeerReviewed text en http://eprints.unisza.edu.my/6049/1/FH02-FSK-19-36067.pdf Rohayu Izanwati, Mohd Rawi and Nurul Shakirah, Azmi and Yow, Weng Kit and Nurul Hana, Zainal Baharin (2017) Expression of beta defensin genes in frozen thawed and cultured immortalised human corneal epithelial cell line. Journal of Biochemistry, Microbiology and Biotechnology, 6 (2). pp. 10-14. ISSN 2289-5779
institution Universiti Sultan Zainal Abidin
building UNISZA Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Sultan Zainal Abidin
content_source UNISZA Institutional Repository
url_provider https://eprints.unisza.edu.my/
language English
topic R Medicine (General)
spellingShingle R Medicine (General)
Rohayu Izanwati, Mohd Rawi
Nurul Shakirah, Azmi
Yow, Weng Kit
Nurul Hana, Zainal Baharin
Expression of beta defensin genes in frozen thawed and cultured immortalised human corneal epithelial cell line
description Human beta defensins (hBD) are important host defence molecules at the ocular surface. In addition to their antimicrobial activities, hBD may also act as regulatory factors in recruiting and activating immune cells. Only hBD-1 – hBD-4 have been well characterised. To date, a complete profile of the beta defensin genes (DEFB) expression in immortalised human corneal epithelial cell line (HCE-2) has not been established. Therefore, this study is aimed to explore a spectrum of DEFB expression in HCE-2. Total RNAs were extracted from frozen thawed HCE-2 and cultured HCE-2 to ensure the gene expression were identical. The RNAs were reverse transcribed into cDNAs. The expression of 10 DEFB (DEFB1, DEFB4A, DEFB103, DEFB104, DEFB105, DEFB106, DEFB109, DEFB123, DEFB126 and DEFB127) were analysed using polymerase chain reaction (PCR) and gel electrophoresis. DEFB1 and DEFB103 were the only hBD mRNAs found constitutively expressed in frozen thawed HCE-2 and cultured HCE-2. It is also interesting to note that PCR enhancer was needed to amplify the genes in cultured HCE-2. Our findings suggest that corneal epithelium constantly produce hBD-1 and hBD-3, which presumably provide the baseline defence against infection. Further investigation on the expression of these genes when HCE-2 stimulated with proinflammatory cytokines would help in better understanding of the ocular surface defence mechanism.
format Article
author Rohayu Izanwati, Mohd Rawi
Nurul Shakirah, Azmi
Yow, Weng Kit
Nurul Hana, Zainal Baharin
author_facet Rohayu Izanwati, Mohd Rawi
Nurul Shakirah, Azmi
Yow, Weng Kit
Nurul Hana, Zainal Baharin
author_sort Rohayu Izanwati, Mohd Rawi
title Expression of beta defensin genes in frozen thawed and cultured immortalised human corneal epithelial cell line
title_short Expression of beta defensin genes in frozen thawed and cultured immortalised human corneal epithelial cell line
title_full Expression of beta defensin genes in frozen thawed and cultured immortalised human corneal epithelial cell line
title_fullStr Expression of beta defensin genes in frozen thawed and cultured immortalised human corneal epithelial cell line
title_full_unstemmed Expression of beta defensin genes in frozen thawed and cultured immortalised human corneal epithelial cell line
title_sort expression of beta defensin genes in frozen thawed and cultured immortalised human corneal epithelial cell line
publishDate 2017
url http://eprints.unisza.edu.my/6049/1/FH02-FSK-19-36067.pdf
http://eprints.unisza.edu.my/6049/
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score 13.211869

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