Expression of Beta Defensin Genes in Frozen Thawed and Cultured Immortalised Human Corneal Epithelial Cell Line

Authors

  • Nurul Shakirah Azmi Medical Genetics Unit, Department of Biomedical Science, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia,43400 UPM Serdang, Selangor Darul Ehsan, Malaysia.
  • Yow Weng Kit Medical Genetics Unit, Department of Biomedical Science, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia,43400 UPM Serdang, Selangor Darul Ehsan, Malaysia
  • Nurul Hana Zainal Baharin Surgery Unit, Department of Medicine, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia,43400 UPM Serdang, Selangor Darul Ehsan, Malaysia
  • Liang Lye Kwan Medical Genetics Unit, Department of Biomedical Science, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia,43400 UPM Serdang, Selangor Darul Ehsan, Malaysia
  • Rohayu Izanwati Mohd Rawi Faculty of Health Sciences, Universiti Sultan Zainal Abidin (UniSZA), Kampus Gong Badak, 21300, Kuala Terengganu, Terengganu, Malaysia.
  • Rafidah Md. Saleh Surgery Unit, Department of Medicine, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia,43400 UPM Serdang, Selangor Darul Ehsan, Malaysia.
  • Nazri Omar Medical Genetics Unit, Department of Biomedical Science, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia,43400 UPM Serdang, Selangor Darul Ehsan, Malaysia.
  • Thilakavathy Karuppiah Genetics and Regenerative Medicine Research Centre, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia,43400 UPM Serdang, Selangor Darul Ehsan, Malaysia. †Deceased 7 January 2018.

DOI:

https://doi.org/10.54987/jobimb.v6i2.427

Keywords:

Beta defensins, DEFB, human corneal epithelium, HCE-2, frozen thawed

Abstract

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.

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Published

31.12.2018

How to Cite

Azmi, N. S., Kit, Y. W., Zainal Baharin, N. H., Kwan, L. L., Mohd Rawi, R. I., Md. Saleh, R., Omar, N., & Karuppiah, T. (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), 10–14. https://doi.org/10.54987/jobimb.v6i2.427

Issue

Vol. 6 No. 2 (2018)

Section

Articles

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