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CIP4 promotes metastasis in triple-negative breast cancer and is associated with poor patient prognosis.

Cerqueira OL, Truesdell P, Baldassarre T, Vilella-Arias SA, Watt K, Meens J, Chander H, Osório CA, Soares FA, Reis EM, Craig AW - Oncotarget (2015)

Bottom Line: This correlated with significant defects in 3D invasion of surrounding extracellular matrix by CIP4 KD TNBC cells when grown as spheroid colonies.In mammary orthotopic xenograft assays using both human TNBC cells (MDA-MB-231, HCC 1806) and rat MTLn3 cells, CIP4 silencing had no overt effect on tumor growth, but significantly reduced the incidence of lung metastases in each tumor model.In human invasive breast cancers, high CIP4 levels was significantly associated with high tumor stage, TNBC and HER2 subtypes, and risk of progression to metastatic disease.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, SP, Brazil.

ABSTRACT
Signaling via epidermal growth factor receptor (EGFR) and Src kinase pathways promote triple-negative breast cancer (TNBC) cell invasion and tumor metastasis. Here, we address the role of Cdc42-interacting protein-4 (CIP4) in TNBC metastasis in vivo, and profile CIP4 expression in human breast cancer patients. In human TNBC cells, CIP4 knock-down (KD) led to less sustained activation of Erk kinase and impaired cell motility compared to control cells. This correlated with significant defects in 3D invasion of surrounding extracellular matrix by CIP4 KD TNBC cells when grown as spheroid colonies. In mammary orthotopic xenograft assays using both human TNBC cells (MDA-MB-231, HCC 1806) and rat MTLn3 cells, CIP4 silencing had no overt effect on tumor growth, but significantly reduced the incidence of lung metastases in each tumor model. In human invasive breast cancers, high CIP4 levels was significantly associated with high tumor stage, TNBC and HER2 subtypes, and risk of progression to metastatic disease. Together, these results implicate CIP4 in promoting metastasis in TNBCs.

No MeSH data available.


Related in: MedlinePlus

CIP4 promotes TNBC cell invasion(A) Representative phase contrast images of spheroid colonies for MDA-MB-231 Tripz or shCIP4 (day 3) and following addition of invasion matrix (day 10, invasion). (B) Graph represents total cell area for spheroids colonies and post-invasion colonies (mean ± SD, *p < 0.05; representative results for 1 of 3 experiments).
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Figure 3: CIP4 promotes TNBC cell invasion(A) Representative phase contrast images of spheroid colonies for MDA-MB-231 Tripz or shCIP4 (day 3) and following addition of invasion matrix (day 10, invasion). (B) Graph represents total cell area for spheroids colonies and post-invasion colonies (mean ± SD, *p < 0.05; representative results for 1 of 3 experiments).

Mentions: Next, we tested the effects of CIP4 silencing on TNBC cell motility and invasion through the ECM. Consistent with the previous study by Pichot et al. using transient CIP4 KD [12], we observed a ~50% reduction in cell motility for Dox-treated shCIP4 cells compared to vector control in cell migration assays (Supplementary Figure S1A). The invasive potential of Dox-treated shCIP4 cells was also analyzed using Transwell™ chambers overlayed with Matrigel™, and we observed a ~75% reduction in numbers of invading cells compared to control cells (Supplementary Figure S1B). We also extended these studies to investigate the effects of CIP4 silencing on cell invasion in 3D assays [22]. In conditions of spheroid formation, CIP4 silencing had no effect on cell growth or size of spheroid colonies (Figure 3A). However, upon addition of ECM supplemented with serum, the shCIP4 spheroids remained compact, and showed reduced invasion through the surrounding ECM compared to that of control spheroid colonies (Figure 3A). Quantification of these results from multiple experimental replicates, revealed a significant reduction in 3D cell invasion upon silencing of CIP4 in TNBC cells (Figure 3B). These results are consistent with our recent study of CIP4 in EGFR-driven lung cancer [21], and support a role for CIP4 in promoting TNBC cell invasion.


CIP4 promotes metastasis in triple-negative breast cancer and is associated with poor patient prognosis.

Cerqueira OL, Truesdell P, Baldassarre T, Vilella-Arias SA, Watt K, Meens J, Chander H, Osório CA, Soares FA, Reis EM, Craig AW - Oncotarget (2015)

CIP4 promotes TNBC cell invasion(A) Representative phase contrast images of spheroid colonies for MDA-MB-231 Tripz or shCIP4 (day 3) and following addition of invasion matrix (day 10, invasion). (B) Graph represents total cell area for spheroids colonies and post-invasion colonies (mean ± SD, *p < 0.05; representative results for 1 of 3 experiments).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4496225&req=5

Figure 3: CIP4 promotes TNBC cell invasion(A) Representative phase contrast images of spheroid colonies for MDA-MB-231 Tripz or shCIP4 (day 3) and following addition of invasion matrix (day 10, invasion). (B) Graph represents total cell area for spheroids colonies and post-invasion colonies (mean ± SD, *p < 0.05; representative results for 1 of 3 experiments).
Mentions: Next, we tested the effects of CIP4 silencing on TNBC cell motility and invasion through the ECM. Consistent with the previous study by Pichot et al. using transient CIP4 KD [12], we observed a ~50% reduction in cell motility for Dox-treated shCIP4 cells compared to vector control in cell migration assays (Supplementary Figure S1A). The invasive potential of Dox-treated shCIP4 cells was also analyzed using Transwell™ chambers overlayed with Matrigel™, and we observed a ~75% reduction in numbers of invading cells compared to control cells (Supplementary Figure S1B). We also extended these studies to investigate the effects of CIP4 silencing on cell invasion in 3D assays [22]. In conditions of spheroid formation, CIP4 silencing had no effect on cell growth or size of spheroid colonies (Figure 3A). However, upon addition of ECM supplemented with serum, the shCIP4 spheroids remained compact, and showed reduced invasion through the surrounding ECM compared to that of control spheroid colonies (Figure 3A). Quantification of these results from multiple experimental replicates, revealed a significant reduction in 3D cell invasion upon silencing of CIP4 in TNBC cells (Figure 3B). These results are consistent with our recent study of CIP4 in EGFR-driven lung cancer [21], and support a role for CIP4 in promoting TNBC cell invasion.

Bottom Line: This correlated with significant defects in 3D invasion of surrounding extracellular matrix by CIP4 KD TNBC cells when grown as spheroid colonies.In mammary orthotopic xenograft assays using both human TNBC cells (MDA-MB-231, HCC 1806) and rat MTLn3 cells, CIP4 silencing had no overt effect on tumor growth, but significantly reduced the incidence of lung metastases in each tumor model.In human invasive breast cancers, high CIP4 levels was significantly associated with high tumor stage, TNBC and HER2 subtypes, and risk of progression to metastatic disease.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, SP, Brazil.

ABSTRACT
Signaling via epidermal growth factor receptor (EGFR) and Src kinase pathways promote triple-negative breast cancer (TNBC) cell invasion and tumor metastasis. Here, we address the role of Cdc42-interacting protein-4 (CIP4) in TNBC metastasis in vivo, and profile CIP4 expression in human breast cancer patients. In human TNBC cells, CIP4 knock-down (KD) led to less sustained activation of Erk kinase and impaired cell motility compared to control cells. This correlated with significant defects in 3D invasion of surrounding extracellular matrix by CIP4 KD TNBC cells when grown as spheroid colonies. In mammary orthotopic xenograft assays using both human TNBC cells (MDA-MB-231, HCC 1806) and rat MTLn3 cells, CIP4 silencing had no overt effect on tumor growth, but significantly reduced the incidence of lung metastases in each tumor model. In human invasive breast cancers, high CIP4 levels was significantly associated with high tumor stage, TNBC and HER2 subtypes, and risk of progression to metastatic disease. Together, these results implicate CIP4 in promoting metastasis in TNBCs.

No MeSH data available.


Related in: MedlinePlus