Cargando…
FAM3B/PANDER inhibits cell death and increases prostate tumor growth by modulating the expression of Bcl-2 and Bcl-X(L) cell survival genes
BACKGROUND: FAM3B/PANDER is a novel cytokine-like protein that induces apoptosis in insulin-secreting beta-cells. Since in silico data revealed that FAM3B can be expressed in prostate tumors, we evaluated the putative role of this cytokine in prostate tumor progression. METHODS: FAM3B expression was...
Autores principales: | , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2018
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5778767/ https://www.ncbi.nlm.nih.gov/pubmed/29357840 http://dx.doi.org/10.1186/s12885-017-3950-9 |
_version_ | 1783294422321987584 |
---|---|
author | Maciel-Silva, Paula Caldeira, Izabela de Assis Santos, Icaro Carreira, Ana Claudia Oliveira Siqueira, Flavia Ramos Antonioli, Eliane Goldberg, Anna Carla Belizário, José Ernesto Garay-Malpartida, Humberto Miguel |
author_facet | Maciel-Silva, Paula Caldeira, Izabela de Assis Santos, Icaro Carreira, Ana Claudia Oliveira Siqueira, Flavia Ramos Antonioli, Eliane Goldberg, Anna Carla Belizário, José Ernesto Garay-Malpartida, Humberto Miguel |
author_sort | Maciel-Silva, Paula |
collection | PubMed |
description | BACKGROUND: FAM3B/PANDER is a novel cytokine-like protein that induces apoptosis in insulin-secreting beta-cells. Since in silico data revealed that FAM3B can be expressed in prostate tumors, we evaluated the putative role of this cytokine in prostate tumor progression. METHODS: FAM3B expression was analyzed by quantitative PCR in tumor tissue clinical samples and prostate tumor cell lines. Culture growth and viability of DU145 cell line were evaluated after treatment with either exogenous FAM3B protein obtained from conditioned media (CM) of 293 T cells overexpressing FAM3B or a recombinant FAM3B protein produced in a bacterial host. DU145 cells overexpressing FAM3B protein were produced by lentiviral-mediated transduction of full-length FAM3B cDNA. Cell viability and apoptosis were analyzed in DU145/FAM3B cells after treatment with several cell death inducers, such as TNF-alpha, staurosporine, etoposide, camptothecin, and serum starvation conditions. Anchorage-independent growth in soft agarose assay was used to evaluate in vitro tumorigenicity. In vivo tumorigenicity and invasiveness were evaluated by tumor xenograft growth in nude mice. RESULTS: We observed an increase in FAM3B expression in prostate tumor samples when compared to normal tissues. DU145 cell viability and survival increased after exogenous treatment with recombinant FAM3B protein or FAM3B-secreted protein. Overexpression of FAM3B in DU145 cells promoted inhibition of DNA fragmentation and phosphatidylserine externalization in a time and dose-dependent fashion, upon apoptosis triggered by TNF-alpha. These events were accompanied by increased gene expression of anti-apoptotic Bcl-2 and Bcl-XL, decreased expression of pro-apoptotic Bax and diminished caspase-3, −8 and −9 proteolytic activities. Furthermore, inhibition of Bcl-2 anti-apoptotic family proteins with small molecules antagonists decreases protective effects of FAM3B in DU145 cells. When compared to the respective controls, cells overexpressing FAM3B displayed a decreased anchorage- independent growth in vitro and increased tumor growth in xenografted nude mice. The immunohistochemistry analysis of tumor xenografts revealed a similar anti-apoptotic phenotype displayed by FAM3B-overexpressing tumor cells. CONCLUSIONS: Taken together, by activating pro-survival mechanisms FAM3B overexpression contributes to increased resistance to cell death and tumor growth in nude mice, highlighting a putative role for this cytokine in prostate cancer progression. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12885-017-3950-9) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-5778767 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-57787672018-01-31 FAM3B/PANDER inhibits cell death and increases prostate tumor growth by modulating the expression of Bcl-2 and Bcl-X(L) cell survival genes Maciel-Silva, Paula Caldeira, Izabela de Assis Santos, Icaro Carreira, Ana Claudia Oliveira Siqueira, Flavia Ramos Antonioli, Eliane Goldberg, Anna Carla Belizário, José Ernesto Garay-Malpartida, Humberto Miguel BMC Cancer Research Article BACKGROUND: FAM3B/PANDER is a novel cytokine-like protein that induces apoptosis in insulin-secreting beta-cells. Since in silico data revealed that FAM3B can be expressed in prostate tumors, we evaluated the putative role of this cytokine in prostate tumor progression. METHODS: FAM3B expression was analyzed by quantitative PCR in tumor tissue clinical samples and prostate tumor cell lines. Culture growth and viability of DU145 cell line were evaluated after treatment with either exogenous FAM3B protein obtained from conditioned media (CM) of 293 T cells overexpressing FAM3B or a recombinant FAM3B protein produced in a bacterial host. DU145 cells overexpressing FAM3B protein were produced by lentiviral-mediated transduction of full-length FAM3B cDNA. Cell viability and apoptosis were analyzed in DU145/FAM3B cells after treatment with several cell death inducers, such as TNF-alpha, staurosporine, etoposide, camptothecin, and serum starvation conditions. Anchorage-independent growth in soft agarose assay was used to evaluate in vitro tumorigenicity. In vivo tumorigenicity and invasiveness were evaluated by tumor xenograft growth in nude mice. RESULTS: We observed an increase in FAM3B expression in prostate tumor samples when compared to normal tissues. DU145 cell viability and survival increased after exogenous treatment with recombinant FAM3B protein or FAM3B-secreted protein. Overexpression of FAM3B in DU145 cells promoted inhibition of DNA fragmentation and phosphatidylserine externalization in a time and dose-dependent fashion, upon apoptosis triggered by TNF-alpha. These events were accompanied by increased gene expression of anti-apoptotic Bcl-2 and Bcl-XL, decreased expression of pro-apoptotic Bax and diminished caspase-3, −8 and −9 proteolytic activities. Furthermore, inhibition of Bcl-2 anti-apoptotic family proteins with small molecules antagonists decreases protective effects of FAM3B in DU145 cells. When compared to the respective controls, cells overexpressing FAM3B displayed a decreased anchorage- independent growth in vitro and increased tumor growth in xenografted nude mice. The immunohistochemistry analysis of tumor xenografts revealed a similar anti-apoptotic phenotype displayed by FAM3B-overexpressing tumor cells. CONCLUSIONS: Taken together, by activating pro-survival mechanisms FAM3B overexpression contributes to increased resistance to cell death and tumor growth in nude mice, highlighting a putative role for this cytokine in prostate cancer progression. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12885-017-3950-9) contains supplementary material, which is available to authorized users. BioMed Central 2018-01-22 /pmc/articles/PMC5778767/ /pubmed/29357840 http://dx.doi.org/10.1186/s12885-017-3950-9 Text en © The Author(s). 2018 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 Article Maciel-Silva, Paula Caldeira, Izabela de Assis Santos, Icaro Carreira, Ana Claudia Oliveira Siqueira, Flavia Ramos Antonioli, Eliane Goldberg, Anna Carla Belizário, José Ernesto Garay-Malpartida, Humberto Miguel FAM3B/PANDER inhibits cell death and increases prostate tumor growth by modulating the expression of Bcl-2 and Bcl-X(L) cell survival genes |
title | FAM3B/PANDER inhibits cell death and increases prostate tumor growth by modulating the expression of Bcl-2 and Bcl-X(L) cell survival genes |
title_full | FAM3B/PANDER inhibits cell death and increases prostate tumor growth by modulating the expression of Bcl-2 and Bcl-X(L) cell survival genes |
title_fullStr | FAM3B/PANDER inhibits cell death and increases prostate tumor growth by modulating the expression of Bcl-2 and Bcl-X(L) cell survival genes |
title_full_unstemmed | FAM3B/PANDER inhibits cell death and increases prostate tumor growth by modulating the expression of Bcl-2 and Bcl-X(L) cell survival genes |
title_short | FAM3B/PANDER inhibits cell death and increases prostate tumor growth by modulating the expression of Bcl-2 and Bcl-X(L) cell survival genes |
title_sort | fam3b/pander inhibits cell death and increases prostate tumor growth by modulating the expression of bcl-2 and bcl-x(l) cell survival genes |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5778767/ https://www.ncbi.nlm.nih.gov/pubmed/29357840 http://dx.doi.org/10.1186/s12885-017-3950-9 |
work_keys_str_mv | AT macielsilvapaula fam3bpanderinhibitscelldeathandincreasesprostatetumorgrowthbymodulatingtheexpressionofbcl2andbclxlcellsurvivalgenes AT caldeiraizabela fam3bpanderinhibitscelldeathandincreasesprostatetumorgrowthbymodulatingtheexpressionofbcl2andbclxlcellsurvivalgenes AT deassissantosicaro fam3bpanderinhibitscelldeathandincreasesprostatetumorgrowthbymodulatingtheexpressionofbcl2andbclxlcellsurvivalgenes AT carreiraanaclaudiaoliveira fam3bpanderinhibitscelldeathandincreasesprostatetumorgrowthbymodulatingtheexpressionofbcl2andbclxlcellsurvivalgenes AT siqueiraflaviaramos fam3bpanderinhibitscelldeathandincreasesprostatetumorgrowthbymodulatingtheexpressionofbcl2andbclxlcellsurvivalgenes AT antoniolieliane fam3bpanderinhibitscelldeathandincreasesprostatetumorgrowthbymodulatingtheexpressionofbcl2andbclxlcellsurvivalgenes AT goldbergannacarla fam3bpanderinhibitscelldeathandincreasesprostatetumorgrowthbymodulatingtheexpressionofbcl2andbclxlcellsurvivalgenes AT belizariojoseernesto fam3bpanderinhibitscelldeathandincreasesprostatetumorgrowthbymodulatingtheexpressionofbcl2andbclxlcellsurvivalgenes AT garaymalpartidahumbertomiguel fam3bpanderinhibitscelldeathandincreasesprostatetumorgrowthbymodulatingtheexpressionofbcl2andbclxlcellsurvivalgenes |