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Immunofluorescent staining and mirna expression of HCT-8 and HT-29 cell lines upon cryptosporidium infection

The protozoan Cryptosporidium mainly infects epithelial cells of the colorectal region. The infection of this species can be observed and confirmed by immunofluorescent staining method in vitro. In addition, the onco-microRNA (miRNA) and tumour suppressor miRNA expression pattern of the epithelial...

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Main Authors: Jainul, Mohammed Abdullah, Md Isa, Muhammad Lokman, Mat Yusof, Afzan
Format: Article
Language:English
English
Published: Malaysian Society of Applied Biology 2018
Subjects:
QH301 Biology
QR Microbiology
RA Public aspects of medicine
Online Access:http://irep.iium.edu.my/70601/1/70601_IMMUNOFLUORESCENT%20STAINING%20AND%20miRNA%20EXPRESSION_article.pdf
http://irep.iium.edu.my/70601/2/70601_IMMUNOFLUORESCENT%20STAINING%20AND%20miRNA%20EXPRESSION_scopus.pdf
http://irep.iium.edu.my/70601/
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spelling my.iium.irep.706012019-02-11T09:30:54Z http://irep.iium.edu.my/70601/ Immunofluorescent staining and mirna expression of HCT-8 and HT-29 cell lines upon cryptosporidium infection Jainul, Mohammed Abdullah Md Isa, Muhammad Lokman Mat Yusof, Afzan QH301 Biology QR Microbiology RA Public aspects of medicine The protozoan Cryptosporidium mainly infects epithelial cells of the colorectal region. The infection of this species can be observed and confirmed by immunofluorescent staining method in vitro. In addition, the onco-microRNA (miRNA) 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. Real Time-quantitative Polymerase Chain Reaction (RT-qPCR) was performed to determine the expression of miR-21 and miR-145 in infected cell lines. Data normalisation 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 normalised 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 onco-miRNAs and tumor suppressor miRNAs. Malaysian Society of Applied Biology 2018-11 Article PeerReviewed application/pdf en http://irep.iium.edu.my/70601/1/70601_IMMUNOFLUORESCENT%20STAINING%20AND%20miRNA%20EXPRESSION_article.pdf application/pdf en http://irep.iium.edu.my/70601/2/70601_IMMUNOFLUORESCENT%20STAINING%20AND%20miRNA%20EXPRESSION_scopus.pdf 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. Malaysian Applied Biology, 47 (5). pp. 101-112. ISSN 0126-8643
institution Universiti Islam Antarabangsa Malaysia
building IIUM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider International Islamic University Malaysia
content_source IIUM Repository (IREP)
url_provider http://irep.iium.edu.my/
language English
English
topic QH301 Biology
QR Microbiology
RA Public aspects of medicine
spellingShingle QH301 Biology
QR Microbiology
RA Public aspects of medicine
Jainul, Mohammed Abdullah
Md Isa, Muhammad Lokman
Mat Yusof, Afzan
Immunofluorescent staining and mirna expression of HCT-8 and HT-29 cell lines upon cryptosporidium infection
description The protozoan Cryptosporidium mainly infects epithelial cells of the colorectal region. The infection of this species can be observed and confirmed by immunofluorescent staining method in vitro. In addition, the onco-microRNA (miRNA) 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. Real Time-quantitative Polymerase Chain Reaction (RT-qPCR) was performed to determine the expression of miR-21 and miR-145 in infected cell lines. Data normalisation 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 normalised 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 onco-miRNAs and tumor suppressor miRNAs.
format Article
author Jainul, Mohammed Abdullah
Md Isa, Muhammad Lokman
Mat Yusof, Afzan
author_facet Jainul, Mohammed Abdullah
Md Isa, Muhammad Lokman
Mat Yusof, Afzan
author_sort Jainul, Mohammed Abdullah
title Immunofluorescent staining and mirna expression of HCT-8 and HT-29 cell lines upon cryptosporidium infection
title_short Immunofluorescent staining and mirna expression of HCT-8 and HT-29 cell lines upon cryptosporidium infection
title_full Immunofluorescent staining and mirna expression of HCT-8 and HT-29 cell lines upon cryptosporidium infection
title_fullStr Immunofluorescent staining and mirna expression of HCT-8 and HT-29 cell lines upon cryptosporidium infection
title_full_unstemmed Immunofluorescent staining and mirna expression of HCT-8 and HT-29 cell lines upon cryptosporidium infection
title_sort immunofluorescent staining and mirna expression of hct-8 and ht-29 cell lines upon cryptosporidium infection
publisher Malaysian Society of Applied Biology
publishDate 2018
url http://irep.iium.edu.my/70601/1/70601_IMMUNOFLUORESCENT%20STAINING%20AND%20miRNA%20EXPRESSION_article.pdf
http://irep.iium.edu.my/70601/2/70601_IMMUNOFLUORESCENT%20STAINING%20AND%20miRNA%20EXPRESSION_scopus.pdf
http://irep.iium.edu.my/70601/
_version_ 1643619240757952512
score 13.211869

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