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

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

Human beta defensins (hBD) are important host defense 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...

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Main Authors:	Azmi, Nurul Shakirah, Yow, Weng Kit, Zainal Baharin, Nurul Hana, Lye, Kwan Liang, Mohd Rawi, Rohayu Izanwati, Md. Saleh, Rafidah, Omar, Nazri, Karuppiah, Thilakavathy
Format:	Article
Language:	English
Published:	Hibiscus Publisher 2018
Online Access:	http://psasir.upm.edu.my/id/eprint/72686/1/Expression%20of%20beta%20defensin%20genes%20in%20frozen%20thawed%20and%20cultured%20immortalised%20human%20corneal%20epithelial%20cell%20line.pdf http://psasir.upm.edu.my/id/eprint/72686/ https://journal.hibiscuspublisher.com/index.php/JOBIMB/article/view/427
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spelling	my.upm.eprints.726862021-01-05T04:47:26Z http://psasir.upm.edu.my/id/eprint/72686/ Expression of beta defensin genes in frozen thawed and cultured immortalised human corneal epithelial cell line Azmi, Nurul Shakirah Yow, Weng Kit Zainal Baharin, Nurul Hana Lye, Kwan Liang Mohd Rawi, Rohayu Izanwati Md. Saleh, Rafidah Omar, Nazri Karuppiah, Thilakavathy Human beta defensins (hBD) are important host defense 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 was 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 defense 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 defense mechanism. Hibiscus Publisher 2018 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/72686/1/Expression%20of%20beta%20defensin%20genes%20in%20frozen%20thawed%20and%20cultured%20immortalised%20human%20corneal%20epithelial%20cell%20line.pdf Azmi, Nurul Shakirah and Yow, Weng Kit and Zainal Baharin, Nurul Hana and Lye, Kwan Liang and Mohd Rawi, Rohayu Izanwati and Md. Saleh, Rafidah and Omar, Nazri and Karuppiah, Thilakavathy (2018) 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 https://journal.hibiscuspublisher.com/index.php/JOBIMB/article/view/427
institution	Universiti Putra Malaysia
building	UPM Library
collection	Institutional Repository
continent	Asia
country	Malaysia
content_provider	Universiti Putra Malaysia
content_source	UPM Institutional Repository
url_provider	http://psasir.upm.edu.my/
language	English
description	Human beta defensins (hBD) are important host defense 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 was 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 defense 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 defense mechanism.
format	Article
author	Azmi, Nurul Shakirah Yow, Weng Kit Zainal Baharin, Nurul Hana Lye, Kwan Liang Mohd Rawi, Rohayu Izanwati Md. Saleh, Rafidah Omar, Nazri Karuppiah, Thilakavathy
spellingShingle	Azmi, Nurul Shakirah Yow, Weng Kit Zainal Baharin, Nurul Hana Lye, Kwan Liang Mohd Rawi, Rohayu Izanwati Md. Saleh, Rafidah Omar, Nazri Karuppiah, Thilakavathy Expression of beta defensin genes in frozen thawed and cultured immortalised human corneal epithelial cell line
author_facet	Azmi, Nurul Shakirah Yow, Weng Kit Zainal Baharin, Nurul Hana Lye, Kwan Liang Mohd Rawi, Rohayu Izanwati Md. Saleh, Rafidah Omar, Nazri Karuppiah, Thilakavathy
author_sort	Azmi, Nurul Shakirah
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
publisher	Hibiscus Publisher
publishDate	2018
url	http://psasir.upm.edu.my/id/eprint/72686/1/Expression%20of%20beta%20defensin%20genes%20in%20frozen%20thawed%20and%20cultured%20immortalised%20human%20corneal%20epithelial%20cell%20line.pdf http://psasir.upm.edu.my/id/eprint/72686/ https://journal.hibiscuspublisher.com/index.php/JOBIMB/article/view/427
_version_	1688549432319016960
score	13.211869

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

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