Calcium Sensor, NCS-1, Promotes Tumor Aggressiveness and Predicts Patient Survival

Moore, Lauren M.; England, Allison; Ehrlich, Barbara E.; Rimm, David L.

doi:10.1158/1541-7786.mcr-16-0408

Cited by 21 publications

(47 citation statements)

References 48 publications

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“…Actin is used as a loading control. Longer exposure shows NCS1 expression in control cells, as previously demonstrated (41). B ) Box plots demonstrating proliferation rates of MDA‐MB231 control and NCS1‐OE cells continuously over 5 d. Circles, triangles, and squares represent different biologic replicates.…”

Section: Resultssupporting

confidence: 86%

“…To explore the in vitro function of NCS1 in malignant tumors, we stably overexpressed NCS1 in MDA‐MB‐231 breast cancer cells (referred to as NCS1‐OE) using a previously described protocol (41). Immunoblotting was performed to confirm successful overexpression of the target gene ( Fig .…”

Section: Resultsmentioning

confidence: 99%

“…ATCC validates all cell lines by Short Tandem Repeat Analysis. The MDA‐MB‐231 cells were transduced with a NCS1 overexpression vector and a control vector as previously described (41). The MDA‐MB‐231 cell lines were maintained at 37°C, 5% CO 2 in DMEM medium supplemented with 10% fetal bovine serum, 1% l ‐glutamine and 1% perdcillin/streptomycin.…”

Section: Methodsmentioning

confidence: 99%

“…Previously (41) generated NCS1‐OE and control MDA‐MB‐231 cells were retro‐virally infected with a triple‐fusion protein reporter. The reporter encodes for herpes simplex virus thymidine kinase 1, green fluorescent protein (GFP) and firefly luciferase (45).…”

Section: Methodsmentioning

confidence: 99%

“…We have previously described NCS1 as a prognostic biomarker in cohorts of breast (41) and liver (42) cancer patients and demonstrated that the overexpression of NCS1 leads to a marked increase in invasion and motility in vitro (41) using 2‐dimensional (2‐D) assays. Furthermore, NCS1 expression levels are highly correlated with other components of Ca 2+ signaling as well as LIMK1 expression (42).…”

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confidence: 99%

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Neuronal calcium sensor 1 (NCS1) promotes motility and metastatic spread of breast cancer cells in vitro and in vivo

et al. 2018

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Increased levels of the calcium‐binding protein neuronal calcium sensor 1 (NCS1) predict an unfavorable patient outcome in several aggressive cancers, including breast and liver tumors. Previous studies suggest that NCS1 overexpression facilitates metastatic spread of these cancers. To investigate this hypothesis, we explored the effects of NCS1 overexpression on cell proliferation, survival, and migration patterns in vitro in 2‐ and 3‐dimensional (2/3‐D). Furthermore, we translated our results into an in vivo mouse xenograft model. Cell‐based proliferation assays were used to demonstrate the effects of overexpression of NCS1 on growth rates. In vitro colony formation and wound healing experiments were performed and 3‐D migration dynamics were studied using collagen gels. Nude mice were injected with breast cancer cells to monitor NCS1‐dependent metastasis formation over time. We observed that increased NCS1 levels do not change cellular growth rates, but do significantly increase 2‐ and 3‐D migration dynamics in vitro. Likewise, NCS1‐overexpressing cells have an increased capacity to form distant metastases and demonstrate better survival and less necrosis in vivo. We found that NCS1 preferentially localizes to the leading edge of cells and overexpression increases the motility of cancer cells. Furthermore, this phenotype is correlated with an increased number of metastases in a xenograft model. These results lay the foundation for exploring the relevance of an NCS1‐mediated pathway as a metastatic biomarker and as a target for pharmacologic interventions.—Apasu, J. E., Schuette, D., LaRanger, R., Steinle, J. A., Nguyen, L. D., Grosshans, H. K., Zhang, M., Cai, W. L., Yan, Q., Robert, M. E., Mak, M., Ehrlich, B. E. Neuronal calcium sensor 1 (NCS1) promotes motility and metastatic spread of breast cancer cells in vitro and in vivo. FASEB J. 33, 4802–4813 (2019). http://www.fasebj.org

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Section: Resultssupporting

confidence: 86%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Neuronal calcium sensor 1 (NCS1) promotes motility and metastatic spread of breast cancer cells in vitro and in vivo

et al. 2018

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NCS‐1 expression is higher in basal breast cancers and regulates calcium influx and cytotoxic responses to doxorubicin

et al. 2019

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Neuronal calcium sensor-1 (NCS-1) is a positive modulator of IP 3 receptors and was recently associated with poorer survival in breast cancers. However, the association between NCS-1 and breast cancer molecular subtypes and the effects of NCS-1 silencing on calcium (Ca 2+ ) signaling in breast cancer cells remain unexplored. Herein, we report for the first time an increased expression of NCS-1 in breast cancers of the basal molecular subtype, a subtype associated with poor prognosis. Using MDA-MB-231 basal breast cancer cells expressing the GCaMP6m Ca 2+ indicator, we showed that NCS-1 silencing did not result in major changes in cytosolic free Ca 2+ increases as a result of endoplasmic reticulum Ca 2+ store mobilization. However, NCS-1 silencing suppressed unstimulated basal Ca 2+ influx. NCS-1 silencing in MDA-MB-231 cells also promoted necrotic cell death induced by the chemotherapeutic drug doxorubicin (1 µM). The effect of NCS-1 silencing on cell death was phenocopied by silencing of ORAI1, a Ca 2+ store-operated Ca 2+ channel that maintains Ca 2+ levels in the endoplasmic reticulum Ca 2+ store and whose expression was significantly positively correlated with NCS-1 in clinical breast cancer samples. This newly identified association between NCS-1 and basal breast cancers, together with the identification of the role of NCS-1 in the regulation of the effects of doxorubicin in MDA-MB-231 breast cancer cells, suggests that NCS-1 and/or pathways regulated by NCS-1 may be important in the treatment of basal breast cancers in women.

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Neuronal Calcium Sensor 1 is up‐regulated in response to stress to promote cell survival and motility in cancer cells

et al. 2020

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Changes in intracellular calcium (Ca 2+ ) signaling can modulate cellular machinery required for cancer progression. Neuronal calcium sensor 1 (NCS1) is a ubiquitously expressed Ca 2+ -binding protein that promotes tumor aggressiveness by enhancing cell survival and metastasis. However, the underlying mechanism by which NCS1 contributes to increased tumor aggressiveness has yet to be identified. In this study, we aimed to determine (a) whether NCS1 expression changes in response to external stimuli, (b) the importance of NCS1 for cell survival and migration, and (c) the cellular mechanism(s) through which NSC1 modulates these outcomes. We found that NCS1 abundance increases under conditions of stress, most prominently after stimulation with the pro-inflammatory cytokine tumor necrosis factor a, in a manner dependent on nuclear factor kappa-light-chain-enhancer of activated B cells (NFjB). We found that NFjB signaling is activated in human breast cancer tissue, which was accompanied by an increase in NCS1 mRNA expression. Further exploration into the relevance of NCS1 in breast cancer progression showed that knockout of NCS1 (NCS1 KO) caused decreased cell survival and motility, increased baseline intracellular Ca 2+ levels, and decreased inositol 1,4,5-trisphosphate-mediated Ca 2+ responses. Protein kinase B (Akt) activity was decreased in NCS1 KO cells, which could be rescued by buffering intracellular Ca 2+ . Conversely, Akt activity was increased in cells overexpressing NCS1 (NCS1 OE). We therefore conclude that NCS1 acts as cellular stress response protein up-regulated by stress-induced NFjB signaling and that NCS1 influences cell survival and motility through effects on Ca 2+ signaling and Akt pathway activation. AbbreviationsAkt, protein kinase B; Ca 2+ , calcium; ER, endoplasmic reticulum; InsP3, inositol 1,4,5-trisphosphate; InsP3R, inositol 1,4,5-trisphosphate receptor; IjBa, nuclear factor of kappa-light polypeptide gene enhancer in B-cell inhibition, alpha; NCS1, neuronal calcium sensor 1; NFjB, nuclear factor kappa-light-chain-enhancer of activated B cells; p-Akt, phosphorylated Akt; PI3K, phosphoinositide-3-kinase; PP2Ac, catalytic subunit of protein phosphatase 2A; tBHP, tert-butylhydroperoxide; TG, thapsigargin; TNF-a, tumor necrosis factor alpha.

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Calcium Sensor, NCS-1, Promotes Tumor Aggressiveness and Predicts Patient Survival

Cited by 21 publications

References 48 publications

Neuronal calcium sensor 1 (NCS1) promotes motility and metastatic spread of breast cancer cells in vitro and in vivo

Neuronal calcium sensor 1 (NCS1) promotes motility and metastatic spread of breast cancer cells in vitro and in vivo

NCS‐1 expression is higher in basal breast cancers and regulates calcium influx and cytotoxic responses to doxorubicin

Neuronal Calcium Sensor 1 is up‐regulated in response to stress to promote cell survival and motility in cancer cells

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