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: | , , , |
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| Format: | Article |
| Language: | English |
| Published: |
2017
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| Subjects: | |
| Online Access: | http://eprints.unisza.edu.my/6049/1/FH02-FSK-19-36067.pdf http://eprints.unisza.edu.my/6049/ |
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| Summary: | 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. |
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