Immunofluorescent staining and miRNA expression of HCT-8 AND HT-29 cell lines upon Cryptosporidium infection

Jainul, Mohammed Abdullah and Md Isa, Muhammad Lokman and Mat Yusof, Afzan (2018) Immunofluorescent staining and miRNA expression of HCT-8 AND HT-29 cell lines upon Cryptosporidium infection. In: The 15th Symposium of Malaysian Society of Applied Biology 2018- Translating Applied Biology for Future Sustainability, 29th June- 1st July 2018, Melaka. (Unpublished)

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Abstract

The protozoan Cryptosporidium mainly infects the epithelial cells of colorectal region. The Infection of this species can be observed and confirmed by immunofluorescent staining method in vitro. Meanwhile, the onco-microRNA and tumour suppressor miRNA expression pattern of the epithelial cells upon Cryptosporidium infection may reveal possible effect of Cryptosporidium in the onset and progression of cancer. In this study, two epithelial colorectal cancer cell lines HCT8 and HT29 were infected with Cryptosporidium parvum (C. parvum) and observed using direct immunofluorescent technique via fluorescein conjugated Vicia Villosa Lectin (VVL) staining. Then, the RT-qPCR was performed to observe the expression of miR-21 and miR-145 in infected cell lines. Data normalization of the miRNA expression was carried out using reference gene RNU44. The immunofluorescent micrographs exhibited the Cryptosporidium infection on both HCT-8 and HT-29 cell lines with green fluorescence. Upregulation of miR-21 and downregulation of normalized expression of miR-145 was observed in both of the cell lines upon Cryptosporidium infection. We found the expression of miR-21 to be increased and miR-145 greatly decreased in both cell lines upon infection. Our current observation suggests that, C. parvum can potentially infect the colorectal cancer cell lines and play critical role in regulating oncomiRNAs and tumor suppressor miRNAs.

Item Type:	Conference or Workshop Item (Plenary Papers)
Additional Information:	5041/67078
Uncontrolled Keywords:	Cryptosporidium parvum, immunofluorescent, RT-qPCR, miRNAs
Subjects:	Q Science > Q Science (General)
Kulliyyahs/Centres/Divisions/Institutes (Can select more than one option. Press CONTROL button):	Kulliyyah of Nursing > Department of Basic Medical Sciences for Nursing Kulliyyah of Nursing
Depositing User:	Dr. Afzan Mat Yusof
Date Deposited:	29 Oct 2018 09:34
Last Modified:	29 Oct 2018 09:34
URI:	http://irep.iium.edu.my/id/eprint/67087

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