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Breast cancer genes PSMC3IP and EPSTI1 play a role in apoptosis regulation.

Capdevila-Busquets E, Badiola N, Arroyo R, Alcalde V, Soler-López M, Aloy P - PLoS ONE (2015)

Bottom Line: A key element to delineate the biology of individual tumors is the regulation of apoptosis.We first explore the existence of direct physical interactions with annotated BC-apoptotic genes.Based on the generated interaction network, we examine several apoptotic markers to determine the effect of PSMC3IP and EPSTI1 gene expression modulation in two different human breast cancer cell lines to suggest potential molecular mechanisms to unveil their role in the disease.

View Article: PubMed Central - PubMed

Affiliation: Joint IRB-BSC-CRG Program in Computational Biology, Institute for Research in Biomedicine (IRB Barcelona), Barcelona, Catalonia, Spain.

ABSTRACT
A key element to delineate the biology of individual tumors is the regulation of apoptosis. In this work, we functionally characterize two breast cancer associated genes, the proteasome 26S subunit ATPase 3 interacting protein (PSMC3IP) and the epithelial-stromal interaction 1 (EPSTI1), to explore their potential apoptotic role in breast cancer. We first explore the existence of direct physical interactions with annotated BC-apoptotic genes. Based on the generated interaction network, we examine several apoptotic markers to determine the effect of PSMC3IP and EPSTI1 gene expression modulation in two different human breast cancer cell lines to suggest potential molecular mechanisms to unveil their role in the disease. Our results show that PSMC3IP and EPSTI1 are able to modulate the extrinsic apoptotic pathway in estrogen receptor positive and triple negative breast cancer cell lines, highlighting them as potential therapeutic targets.

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Modulation of PSMC3IP and EPSTI1 expression in breast cancer cells.(A-B) Genes were overexpressed as Myc-tagged fusion proteins in different cell lines and protein relative levels were analysed based on MYC-tag empty transfection vector (Vector). (C-D) Endogenous gene expression was silenced using specific siRNA and depletion levels were analysed based on siRNA against luciferase expression (siLUC) as a negative control. Prior to depletion experiments, EPSTI1 expression was induced by treating cells with IFN-α at 1000 U/ml for 8h. XIAP was used as a reference anti-apoptotic protein in all experiments. Each bar represents the mean ±SD of three experiments performed in duplicate (*P <0.05, **P <0.01, ***P <0.001 vs MYC-tag vector in overexpression assays and vs siLUCIFERASE in silencing).
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pone.0115352.g003: Modulation of PSMC3IP and EPSTI1 expression in breast cancer cells.(A-B) Genes were overexpressed as Myc-tagged fusion proteins in different cell lines and protein relative levels were analysed based on MYC-tag empty transfection vector (Vector). (C-D) Endogenous gene expression was silenced using specific siRNA and depletion levels were analysed based on siRNA against luciferase expression (siLUC) as a negative control. Prior to depletion experiments, EPSTI1 expression was induced by treating cells with IFN-α at 1000 U/ml for 8h. XIAP was used as a reference anti-apoptotic protein in all experiments. Each bar represents the mean ±SD of three experiments performed in duplicate (*P <0.05, **P <0.01, ***P <0.001 vs MYC-tag vector in overexpression assays and vs siLUCIFERASE in silencing).

Mentions: We then modulated PSMC3IP and EPSTI1 differential expression by either cDNA or siRNA knockout cell transfection. We included the X-linked inhibitor of apoptosis protein (XIAP) as an anti-apoptotic reference gene, since it is a well-characterized inhibitor of caspase-3, caspase-7 and caspase-9 [40, 41] (Fig. 3). Compared to cells transfected with empty vectors, we observed a highly overexpression of PSMC3IP in both cell lines (MDA-MB-231, 6.5-fold; MCF-7, 13-fold) (Fig. 3A–3B), albeit we only achieved a moderate EPSTI1 overexpression (2.1-fold and 2.6-fold, respectively (Fig. 3A–3B). On the other hand, by siRNA transfection, we were able to reduce PSMC3IP levels by 70% in MDA-MB-231 and 50% in MCF-7 cells compared to cells transfected with control siRNA (siLUC) (Fig. 3C–3D). To maximize and visualize the effect of EPSTI1 depletion, we induced its endogenous expression with IFN-α prior to gene silencing in both cell lines (Fig. 3C–3D), as previously reported [17].


Breast cancer genes PSMC3IP and EPSTI1 play a role in apoptosis regulation.

Capdevila-Busquets E, Badiola N, Arroyo R, Alcalde V, Soler-López M, Aloy P - PLoS ONE (2015)

Modulation of PSMC3IP and EPSTI1 expression in breast cancer cells.(A-B) Genes were overexpressed as Myc-tagged fusion proteins in different cell lines and protein relative levels were analysed based on MYC-tag empty transfection vector (Vector). (C-D) Endogenous gene expression was silenced using specific siRNA and depletion levels were analysed based on siRNA against luciferase expression (siLUC) as a negative control. Prior to depletion experiments, EPSTI1 expression was induced by treating cells with IFN-α at 1000 U/ml for 8h. XIAP was used as a reference anti-apoptotic protein in all experiments. Each bar represents the mean ±SD of three experiments performed in duplicate (*P <0.05, **P <0.01, ***P <0.001 vs MYC-tag vector in overexpression assays and vs siLUCIFERASE in silencing).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4295872&req=5

pone.0115352.g003: Modulation of PSMC3IP and EPSTI1 expression in breast cancer cells.(A-B) Genes were overexpressed as Myc-tagged fusion proteins in different cell lines and protein relative levels were analysed based on MYC-tag empty transfection vector (Vector). (C-D) Endogenous gene expression was silenced using specific siRNA and depletion levels were analysed based on siRNA against luciferase expression (siLUC) as a negative control. Prior to depletion experiments, EPSTI1 expression was induced by treating cells with IFN-α at 1000 U/ml for 8h. XIAP was used as a reference anti-apoptotic protein in all experiments. Each bar represents the mean ±SD of three experiments performed in duplicate (*P <0.05, **P <0.01, ***P <0.001 vs MYC-tag vector in overexpression assays and vs siLUCIFERASE in silencing).
Mentions: We then modulated PSMC3IP and EPSTI1 differential expression by either cDNA or siRNA knockout cell transfection. We included the X-linked inhibitor of apoptosis protein (XIAP) as an anti-apoptotic reference gene, since it is a well-characterized inhibitor of caspase-3, caspase-7 and caspase-9 [40, 41] (Fig. 3). Compared to cells transfected with empty vectors, we observed a highly overexpression of PSMC3IP in both cell lines (MDA-MB-231, 6.5-fold; MCF-7, 13-fold) (Fig. 3A–3B), albeit we only achieved a moderate EPSTI1 overexpression (2.1-fold and 2.6-fold, respectively (Fig. 3A–3B). On the other hand, by siRNA transfection, we were able to reduce PSMC3IP levels by 70% in MDA-MB-231 and 50% in MCF-7 cells compared to cells transfected with control siRNA (siLUC) (Fig. 3C–3D). To maximize and visualize the effect of EPSTI1 depletion, we induced its endogenous expression with IFN-α prior to gene silencing in both cell lines (Fig. 3C–3D), as previously reported [17].

Bottom Line: A key element to delineate the biology of individual tumors is the regulation of apoptosis.We first explore the existence of direct physical interactions with annotated BC-apoptotic genes.Based on the generated interaction network, we examine several apoptotic markers to determine the effect of PSMC3IP and EPSTI1 gene expression modulation in two different human breast cancer cell lines to suggest potential molecular mechanisms to unveil their role in the disease.

View Article: PubMed Central - PubMed

Affiliation: Joint IRB-BSC-CRG Program in Computational Biology, Institute for Research in Biomedicine (IRB Barcelona), Barcelona, Catalonia, Spain.

ABSTRACT
A key element to delineate the biology of individual tumors is the regulation of apoptosis. In this work, we functionally characterize two breast cancer associated genes, the proteasome 26S subunit ATPase 3 interacting protein (PSMC3IP) and the epithelial-stromal interaction 1 (EPSTI1), to explore their potential apoptotic role in breast cancer. We first explore the existence of direct physical interactions with annotated BC-apoptotic genes. Based on the generated interaction network, we examine several apoptotic markers to determine the effect of PSMC3IP and EPSTI1 gene expression modulation in two different human breast cancer cell lines to suggest potential molecular mechanisms to unveil their role in the disease. Our results show that PSMC3IP and EPSTI1 are able to modulate the extrinsic apoptotic pathway in estrogen receptor positive and triple negative breast cancer cell lines, highlighting them as potential therapeutic targets.

Show MeSH
Related in: MedlinePlus