Cargando…

Rad51C-ATXN7 fusion gene expression in colorectal tumors

BACKGROUND: Fusion proteins have unique oncogenic properties and their identification can be useful either as diagnostic or therapeutic targets. Next generation sequencing data have previously shown a fusion gene formed between Rad51C and ATXN7 genes in the MCF7 breast cancer cell line. However, the...

Descripción completa

Detalles Bibliográficos
Autores principales: Kalvala, Arjun, Gao, Li, Aguila, Brittany, Dotts, Kathleen, Rahman, Mohammad, Nana-Sinkam, Serge P., Zhou, Xiaoping, Wang, Qi-En, Amann, Joseph, Otterson, Gregory A., Villalona-Calero, Miguel A., Duan, Wenrui
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4906819/
https://www.ncbi.nlm.nih.gov/pubmed/27296891
http://dx.doi.org/10.1186/s12943-016-0527-1
_version_ 1782437474108178432
author Kalvala, Arjun
Gao, Li
Aguila, Brittany
Dotts, Kathleen
Rahman, Mohammad
Nana-Sinkam, Serge P.
Zhou, Xiaoping
Wang, Qi-En
Amann, Joseph
Otterson, Gregory A.
Villalona-Calero, Miguel A.
Duan, Wenrui
author_facet Kalvala, Arjun
Gao, Li
Aguila, Brittany
Dotts, Kathleen
Rahman, Mohammad
Nana-Sinkam, Serge P.
Zhou, Xiaoping
Wang, Qi-En
Amann, Joseph
Otterson, Gregory A.
Villalona-Calero, Miguel A.
Duan, Wenrui
author_sort Kalvala, Arjun
collection PubMed
description BACKGROUND: Fusion proteins have unique oncogenic properties and their identification can be useful either as diagnostic or therapeutic targets. Next generation sequencing data have previously shown a fusion gene formed between Rad51C and ATXN7 genes in the MCF7 breast cancer cell line. However, the existence of this fusion gene in colorectal patient tumor tissues is largely still unknown. METHODS: We evaluated for the presence of Rad51C-ATXN7 fusion gene in colorectal tumors and cells by RT-PCR, PCR, Topo TA cloning, Real time PCR, immunoprecipitation and immunoblotting techniques. RESULTS: We identified two forms of fusion mRNAs between Rad51C and ATXN7 in the colorectal tumors, including a Variant 1 (fusion transcript between Rad51C exons 1–7 and ATXN7 exons 6–13), and a Variant 2 (between Rad51C exons 1–6 and ATXN7 exons 6–13). In silico analysis showed that the Variant 1 produces a truncated protein, whereas the Variant 2 was predicted to produce a fusion protein with molecular weight of 110 KDa. Immunoprecipitation and Western blot analysis further showed a 110 KDa protein in colorectal tumors. 5-Azacytidine treatment of LS-174 T cells caused a 3.51-fold increase in expression of the fusion gene (Variant 2) as compared to no treatment controls evaluated by real time PCR. CONCLUSION: In conclusion we found a fusion gene between DNA repair gene Rad51C and neuro-cerebral ataxia Ataxin-7 gene in colorectal tumors. The in-frame fusion transcript of Variant 2 results in a fusion protein with molecular weight of 110 KDa. In addition, we found that expression of fusion gene is associated with functional impairment of Fanconi Anemia (FA) DNA repair pathway in colorectal tumors. The expression of Rad51C-ATXN7 in tumors warrants further investigation, as it suggests the potential of the fusion gene in treatment and predictive value in colorectal cancers. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12943-016-0527-1) contains supplementary material, which is available to authorized users.
format Online
Article
Text
id pubmed-4906819
institution National Center for Biotechnology Information
language English
publishDate 2016
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-49068192016-06-15 Rad51C-ATXN7 fusion gene expression in colorectal tumors Kalvala, Arjun Gao, Li Aguila, Brittany Dotts, Kathleen Rahman, Mohammad Nana-Sinkam, Serge P. Zhou, Xiaoping Wang, Qi-En Amann, Joseph Otterson, Gregory A. Villalona-Calero, Miguel A. Duan, Wenrui Mol Cancer Research BACKGROUND: Fusion proteins have unique oncogenic properties and their identification can be useful either as diagnostic or therapeutic targets. Next generation sequencing data have previously shown a fusion gene formed between Rad51C and ATXN7 genes in the MCF7 breast cancer cell line. However, the existence of this fusion gene in colorectal patient tumor tissues is largely still unknown. METHODS: We evaluated for the presence of Rad51C-ATXN7 fusion gene in colorectal tumors and cells by RT-PCR, PCR, Topo TA cloning, Real time PCR, immunoprecipitation and immunoblotting techniques. RESULTS: We identified two forms of fusion mRNAs between Rad51C and ATXN7 in the colorectal tumors, including a Variant 1 (fusion transcript between Rad51C exons 1–7 and ATXN7 exons 6–13), and a Variant 2 (between Rad51C exons 1–6 and ATXN7 exons 6–13). In silico analysis showed that the Variant 1 produces a truncated protein, whereas the Variant 2 was predicted to produce a fusion protein with molecular weight of 110 KDa. Immunoprecipitation and Western blot analysis further showed a 110 KDa protein in colorectal tumors. 5-Azacytidine treatment of LS-174 T cells caused a 3.51-fold increase in expression of the fusion gene (Variant 2) as compared to no treatment controls evaluated by real time PCR. CONCLUSION: In conclusion we found a fusion gene between DNA repair gene Rad51C and neuro-cerebral ataxia Ataxin-7 gene in colorectal tumors. The in-frame fusion transcript of Variant 2 results in a fusion protein with molecular weight of 110 KDa. In addition, we found that expression of fusion gene is associated with functional impairment of Fanconi Anemia (FA) DNA repair pathway in colorectal tumors. The expression of Rad51C-ATXN7 in tumors warrants further investigation, as it suggests the potential of the fusion gene in treatment and predictive value in colorectal cancers. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12943-016-0527-1) contains supplementary material, which is available to authorized users. BioMed Central 2016-06-13 /pmc/articles/PMC4906819/ /pubmed/27296891 http://dx.doi.org/10.1186/s12943-016-0527-1 Text en © Kalvala et al. 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. 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 Research
Kalvala, Arjun
Gao, Li
Aguila, Brittany
Dotts, Kathleen
Rahman, Mohammad
Nana-Sinkam, Serge P.
Zhou, Xiaoping
Wang, Qi-En
Amann, Joseph
Otterson, Gregory A.
Villalona-Calero, Miguel A.
Duan, Wenrui
Rad51C-ATXN7 fusion gene expression in colorectal tumors
title Rad51C-ATXN7 fusion gene expression in colorectal tumors
title_full Rad51C-ATXN7 fusion gene expression in colorectal tumors
title_fullStr Rad51C-ATXN7 fusion gene expression in colorectal tumors
title_full_unstemmed Rad51C-ATXN7 fusion gene expression in colorectal tumors
title_short Rad51C-ATXN7 fusion gene expression in colorectal tumors
title_sort rad51c-atxn7 fusion gene expression in colorectal tumors
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4906819/
https://www.ncbi.nlm.nih.gov/pubmed/27296891
http://dx.doi.org/10.1186/s12943-016-0527-1
work_keys_str_mv AT kalvalaarjun rad51catxn7fusiongeneexpressionincolorectaltumors
AT gaoli rad51catxn7fusiongeneexpressionincolorectaltumors
AT aguilabrittany rad51catxn7fusiongeneexpressionincolorectaltumors
AT dottskathleen rad51catxn7fusiongeneexpressionincolorectaltumors
AT rahmanmohammad rad51catxn7fusiongeneexpressionincolorectaltumors
AT nanasinkamsergep rad51catxn7fusiongeneexpressionincolorectaltumors
AT zhouxiaoping rad51catxn7fusiongeneexpressionincolorectaltumors
AT wangqien rad51catxn7fusiongeneexpressionincolorectaltumors
AT amannjoseph rad51catxn7fusiongeneexpressionincolorectaltumors
AT ottersongregorya rad51catxn7fusiongeneexpressionincolorectaltumors
AT villalonacaleromiguela rad51catxn7fusiongeneexpressionincolorectaltumors
AT duanwenrui rad51catxn7fusiongeneexpressionincolorectaltumors