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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 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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Główni autorzy: 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: Artykuł
Język:English
Wydane: Hibiscus Publisher 2018
Dostęp online: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
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spelling oai:psasir.upm.edu.my:72686 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 UPM IR
collection UPM IR
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
_version_ 1782758159492841472
score 12.933938

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