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Observations on the expression of human papillomavirus major capsid protein in HeLa cells
BACKGROUND: The goal of this study was to identify the nature of the inclusion bodies that have been found in HeLa cells (cervical cancer immortal cell line) by electron microscope and to determine whether the major capsid protein (L1) of human papillomavirus (HPV) can be expressed in HPV-positive u...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4461987/ https://www.ncbi.nlm.nih.gov/pubmed/26064080 http://dx.doi.org/10.1186/s12935-015-0206-0 |
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author | Xiao, Chang-Yi Fu, Bing-Bing Li, Zhi-Ying Mushtaq, Gohar Kamal, Mohammad Amjad Li, Jia-Hua Tang, Gui-Cheng Xiao, Shuo-Shuang |
author_facet | Xiao, Chang-Yi Fu, Bing-Bing Li, Zhi-Ying Mushtaq, Gohar Kamal, Mohammad Amjad Li, Jia-Hua Tang, Gui-Cheng Xiao, Shuo-Shuang |
author_sort | Xiao, Chang-Yi |
collection | PubMed |
description | BACKGROUND: The goal of this study was to identify the nature of the inclusion bodies that have been found in HeLa cells (cervical cancer immortal cell line) by electron microscope and to determine whether the major capsid protein (L1) of human papillomavirus (HPV) can be expressed in HPV-positive uterine cervix cancer cells. METHODS: HPV L1 protein expression in HeLa cells was detected with anti-HPV L1 multivalent mice monoclonal antibody and rabbit polyclonal anti-HPV L1 antibody by ELISA, light microscope immunohistochemistry, electron microscope immunocytochemistry and Western blotting assays. Reverse transcriptional PCR (RT-PCR) was performed to detect the transcription of L1 mRNA in HeLa cells. The immortalized human keratinocyte HeCat was used as the negative control. RESULTS: HPV L1 proteins reacted positively in the lysate of HeLa cells by ELISA assays. HRP labeled light microscope immunohistochemistry assay showed that there was a strong HPV L1 positive reaction in HeLa cells. Under the electron microscope, irregular shaped inclusion bodies, assembled by many small and uniform granules, had been observed in the cytoplasm of some HeLa cells. These granules could be labeled by the colloidal gold carried by HPV L1 antibody. The Western blotting assay showed that there was a L1 reaction strap at 80–85 kDa in the HeLa cell lysates, hence demonstrating the existence of HPV18 L1 in HeLa cells. RT-PCR assay showed that the L1 mRNA was transcribed in HeLa cells. CONCLUSIONS: The inclusion bodies found in the cytoplasm of HeLa cells are composed of HPV18 L1 protein. Since HeLa cell line is a type of cervical cancer cells, this implies that HeLa cells have the ability to express HPV L1 proteins. |
format | Online Article Text |
id | pubmed-4461987 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-44619872015-06-11 Observations on the expression of human papillomavirus major capsid protein in HeLa cells Xiao, Chang-Yi Fu, Bing-Bing Li, Zhi-Ying Mushtaq, Gohar Kamal, Mohammad Amjad Li, Jia-Hua Tang, Gui-Cheng Xiao, Shuo-Shuang Cancer Cell Int Primary Research BACKGROUND: The goal of this study was to identify the nature of the inclusion bodies that have been found in HeLa cells (cervical cancer immortal cell line) by electron microscope and to determine whether the major capsid protein (L1) of human papillomavirus (HPV) can be expressed in HPV-positive uterine cervix cancer cells. METHODS: HPV L1 protein expression in HeLa cells was detected with anti-HPV L1 multivalent mice monoclonal antibody and rabbit polyclonal anti-HPV L1 antibody by ELISA, light microscope immunohistochemistry, electron microscope immunocytochemistry and Western blotting assays. Reverse transcriptional PCR (RT-PCR) was performed to detect the transcription of L1 mRNA in HeLa cells. The immortalized human keratinocyte HeCat was used as the negative control. RESULTS: HPV L1 proteins reacted positively in the lysate of HeLa cells by ELISA assays. HRP labeled light microscope immunohistochemistry assay showed that there was a strong HPV L1 positive reaction in HeLa cells. Under the electron microscope, irregular shaped inclusion bodies, assembled by many small and uniform granules, had been observed in the cytoplasm of some HeLa cells. These granules could be labeled by the colloidal gold carried by HPV L1 antibody. The Western blotting assay showed that there was a L1 reaction strap at 80–85 kDa in the HeLa cell lysates, hence demonstrating the existence of HPV18 L1 in HeLa cells. RT-PCR assay showed that the L1 mRNA was transcribed in HeLa cells. CONCLUSIONS: The inclusion bodies found in the cytoplasm of HeLa cells are composed of HPV18 L1 protein. Since HeLa cell line is a type of cervical cancer cells, this implies that HeLa cells have the ability to express HPV L1 proteins. BioMed Central 2015-05-23 /pmc/articles/PMC4461987/ /pubmed/26064080 http://dx.doi.org/10.1186/s12935-015-0206-0 Text en © Xiao et al. 2015 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Primary Research Xiao, Chang-Yi Fu, Bing-Bing Li, Zhi-Ying Mushtaq, Gohar Kamal, Mohammad Amjad Li, Jia-Hua Tang, Gui-Cheng Xiao, Shuo-Shuang Observations on the expression of human papillomavirus major capsid protein in HeLa cells |
title | Observations on the expression of human papillomavirus major capsid protein in HeLa cells |
title_full | Observations on the expression of human papillomavirus major capsid protein in HeLa cells |
title_fullStr | Observations on the expression of human papillomavirus major capsid protein in HeLa cells |
title_full_unstemmed | Observations on the expression of human papillomavirus major capsid protein in HeLa cells |
title_short | Observations on the expression of human papillomavirus major capsid protein in HeLa cells |
title_sort | observations on the expression of human papillomavirus major capsid protein in hela cells |
topic | Primary Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4461987/ https://www.ncbi.nlm.nih.gov/pubmed/26064080 http://dx.doi.org/10.1186/s12935-015-0206-0 |
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