Overexpression of CA1 mRNA and the CA I Protein in Tumor Cells Does Not Change the Gene Expression of the ECM Proteins

In our study, we performed retroviral transduction to overexpress codon-optimized variant of gene encoding human carbonic anhydrase I (optiCA1) in two tumor cell lines PC3 and MDA-MB-231, derived from human prostatic and breast carcinoma respectively. We achieved significantly enhanced and stable ov...

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Main Authors: Ján Lakota, Mária Dubrovčáková
Format: Article
Language:English
Published: MDPI AG 2020-01-01
Series:International Journal of Molecular Sciences
Subjects:
Online Access:https://www.mdpi.com/1422-0067/21/2/639
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spelling doaj-befc86370eb64dd9b6f8ea78620306332020-11-25T02:30:03ZengMDPI AGInternational Journal of Molecular Sciences1422-00672020-01-0121263910.3390/ijms21020639ijms21020639Overexpression of CA1 mRNA and the CA I Protein in Tumor Cells Does Not Change the Gene Expression of the ECM ProteinsJán Lakota0Mária Dubrovčáková1Biomedical Research Center, SAS, Dubravska cesta 9, 845 05 Bratislava, SlovakiaBiomedical Research Center, SAS, Dubravska cesta 9, 845 05 Bratislava, SlovakiaIn our study, we performed retroviral transduction to overexpress codon-optimized variant of gene encoding human carbonic anhydrase I (optiCA1) in two tumor cell lines PC3 and MDA-MB-231, derived from human prostatic and breast carcinoma respectively. We achieved significantly enhanced and stable overexpression of exogenous optiCA1 gene. The expression of endogenous, wild CA1 gene was found to be normally low (C<sub>t</sub> 28.6 for PC3 cells) or below to the detection limit (C<sub>t</sub> 35.5 for MDA-MB-231 cells). No morphological changes and no decreasing viability of tumor cells were observed upon stable overexpression of the optiCA1 gene. In our study we have shown that the overexpression of the optimized human CA1 in engineered PC3 and MDA-MB-231 cells did not induce similar changes as we observed in tumor cells cultivated in the presence of human sera containing extensively high titers of anti-CA I autoantibodies from patients with complete remission of malignant disease. In both optiCA1transduced cell lines, the expression of selected genes responsible for basal lamina assembly, cytoskeleton, extracellular matrix proteins and proto-oncogenes (COL1A1, COL4A4, LAMC2, CTHRC1, and WNT7B) was not changed.https://www.mdpi.com/1422-0067/21/2/639carbonic anhydrase ica1 gene overexpressiontumor cell
collection DOAJ
language English
format Article
sources DOAJ
author Ján Lakota
Mária Dubrovčáková
spellingShingle Ján Lakota
Mária Dubrovčáková
Overexpression of CA1 mRNA and the CA I Protein in Tumor Cells Does Not Change the Gene Expression of the ECM Proteins
International Journal of Molecular Sciences
carbonic anhydrase i
ca1 gene overexpression
tumor cell
author_facet Ján Lakota
Mária Dubrovčáková
author_sort Ján Lakota
title Overexpression of CA1 mRNA and the CA I Protein in Tumor Cells Does Not Change the Gene Expression of the ECM Proteins
title_short Overexpression of CA1 mRNA and the CA I Protein in Tumor Cells Does Not Change the Gene Expression of the ECM Proteins
title_full Overexpression of CA1 mRNA and the CA I Protein in Tumor Cells Does Not Change the Gene Expression of the ECM Proteins
title_fullStr Overexpression of CA1 mRNA and the CA I Protein in Tumor Cells Does Not Change the Gene Expression of the ECM Proteins
title_full_unstemmed Overexpression of CA1 mRNA and the CA I Protein in Tumor Cells Does Not Change the Gene Expression of the ECM Proteins
title_sort overexpression of ca1 mrna and the ca i protein in tumor cells does not change the gene expression of the ecm proteins
publisher MDPI AG
series International Journal of Molecular Sciences
issn 1422-0067
publishDate 2020-01-01
description In our study, we performed retroviral transduction to overexpress codon-optimized variant of gene encoding human carbonic anhydrase I (optiCA1) in two tumor cell lines PC3 and MDA-MB-231, derived from human prostatic and breast carcinoma respectively. We achieved significantly enhanced and stable overexpression of exogenous optiCA1 gene. The expression of endogenous, wild CA1 gene was found to be normally low (C<sub>t</sub> 28.6 for PC3 cells) or below to the detection limit (C<sub>t</sub> 35.5 for MDA-MB-231 cells). No morphological changes and no decreasing viability of tumor cells were observed upon stable overexpression of the optiCA1 gene. In our study we have shown that the overexpression of the optimized human CA1 in engineered PC3 and MDA-MB-231 cells did not induce similar changes as we observed in tumor cells cultivated in the presence of human sera containing extensively high titers of anti-CA I autoantibodies from patients with complete remission of malignant disease. In both optiCA1transduced cell lines, the expression of selected genes responsible for basal lamina assembly, cytoskeleton, extracellular matrix proteins and proto-oncogenes (COL1A1, COL4A4, LAMC2, CTHRC1, and WNT7B) was not changed.
topic carbonic anhydrase i
ca1 gene overexpression
tumor cell
url https://www.mdpi.com/1422-0067/21/2/639
work_keys_str_mv AT janlakota overexpressionofca1mrnaandthecaiproteinintumorcellsdoesnotchangethegeneexpressionoftheecmproteins
AT mariadubrovcakova overexpressionofca1mrnaandthecaiproteinintumorcellsdoesnotchangethegeneexpressionoftheecmproteins
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