PMID- 31652779
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20200928
IS  - 2072-6694 (Print)
IS  - 2072-6694 (Electronic)
IS  - 2072-6694 (Linking)
VI  - 11
IP  - 11
DP  - 2019 Oct 23
TI  - Adenylyl Cyclase Type 8 Overexpression Impairs Phosphorylation-Dependent Orai1 
      Inactivation and Promotes Migration in MDA-MB-231 Breast Cancer Cells.
LID - 10.3390/cancers11111624 [doi]
LID - 1624
AB  - Orai1 plays a major role in store-operated Ca(2+) entry (SOCE) in triple-negative 
      breast cancer (TNBC) cells. This channel is inactivated via different mechanisms, 
      including protein kinase C (PKC) and protein kinase A (PKA)-dependent 
      phosphorylation at Ser-27 and Ser-30 or Ser-34, respectively, which shapes the 
      Ca(2+) responses to agonists. The Ca(2+) calmodulin-activated adenylyl cyclase 
      type 8 (AC8) was reported to interact directly with Orai1, thus mediating a 
      dynamic interplay between the Ca(2+)- and cyclic adenosine monophosphate 
      (cAMP)-dependent signaling pathways. Here, we show that the breast cancer cell 
      lines MCF7 and MDA-MB-231 exhibit enhanced expression of Orai1 and AC8 as 
      compared to the non-tumoral breast epithelial MCF10A cell line. In these cells, 
      AC8 interacts with the Orai1alpha variant in a manner that is not regulated by Orai1 
      phosphorylation. AC8 knockdown in MDA-MB-231 cells, using two different small 
      interfering RNAs (siRNAs), attenuates thapsigargin (TG)-induced Ca(2+) entry and 
      also Ca(2+) influx mediated by co-expression of Orai1 and the Orai1-activating 
      small fragment (OASF) of STIM1 (stromal interaction molecule-1). Conversely, AC8 
      overexpression enhances SOCE, as well as Ca(2+) entry, in cells co-expressing 
      Orai1 and OASF. In MDA-MB-231 cells, we found that AC8 overexpression reduces the 
      Orai1 phosphoserine content, thus suggesting that AC8 interferes with Orai1 
      serine phosphorylation, which takes place at residues located in the AC8-binding 
      site. Consistent with this, the subset of Orai1 associated with AC8 in naive 
      MDA-MB-231 cells is not phosphorylated in serine residues in contrast to the 
      AC8-independent Orai1 subset. AC8 expression knockdown attenuates migration of 
      MCF7 and MDA-MB-231 cells, while this maneuver has no effect in the MCF10A cell 
      line, which is likely attributed to the low expression of AC8 in these cells. We 
      found that AC8 is required for FAK (focal adhesion kinase) phosphorylation in 
      MDA-MB-231 cells, which might explain its role in cell migration. Finally, we 
      found that AC8 is required for TNBC cell proliferation. These findings indicate 
      that overexpression of AC8 in breast cancer MDA-MB-231 cells impairs the 
      phosphorylation-dependent Orai1 inactivation, a mechanism that might support the 
      enhanced ability of these cells to migrate.
FAU - Sanchez-Collado, Jose
AU  - Sanchez-Collado J
AD  - Department of Physiology, (Cellular Physiology Research Group), Institute of 
      Molecular Pathology Biomarkers, University of Extremadura, 10003 Caceres, Spain. 
      josesc@unex.es.
FAU - Lopez, Jose J
AU  - Lopez JJ
AD  - Department of Physiology, (Cellular Physiology Research Group), Institute of 
      Molecular Pathology Biomarkers, University of Extremadura, 10003 Caceres, Spain. 
      jjlopez@unex.es.
FAU - Jardin, Isaac
AU  - Jardin I
AD  - Department of Physiology, (Cellular Physiology Research Group), Institute of 
      Molecular Pathology Biomarkers, University of Extremadura, 10003 Caceres, Spain. 
      ijp@unex.es.
FAU - Camello, Pedro J
AU  - Camello PJ
AD  - Department of Physiology, (Smooth Muscle Physiology Research Group), Institute of 
      Molecular Pathology Biomarkers, University of Extremadura, 10003 Caceres, Spain. 
      pcamello@unex.es.
FAU - Falcon, Debora
AU  - Falcon D
AD  - Department of Medical Physiology and Biophysics, Institute of Biomedicine of 
      Sevilla, 41013 Sevilla, Spain. dfalboy@gmail.com.
FAU - Regodon, Sergio
AU  - Regodon S
AD  - Department of Physiology, (Cellular Physiology Research Group), Institute of 
      Molecular Pathology Biomarkers, University of Extremadura, 10003 Caceres, Spain. 
      sergio@unex.es.
FAU - Salido, Gines M
AU  - Salido GM
AD  - Department of Physiology, (Cellular Physiology Research Group), Institute of 
      Molecular Pathology Biomarkers, University of Extremadura, 10003 Caceres, Spain. 
      gsalido@unex.es.
FAU - Smani, Tarik
AU  - Smani T
AD  - Department of Medical Physiology and Biophysics, Institute of Biomedicine of 
      Sevilla, 41013 Sevilla, Spain. tasmani@us.es.
FAU - Rosado, Juan A
AU  - Rosado JA
AD  - Department of Physiology, (Cellular Physiology Research Group), Institute of 
      Molecular Pathology Biomarkers, University of Extremadura, 10003 Caceres, Spain. 
      jarosado@unex.es.
LA  - eng
PT  - Journal Article
DEP - 20191023
PL  - Switzerland
TA  - Cancers (Basel)
JT  - Cancers
JID - 101526829
PMC - PMC6893434
OTO - NOTNLM
OT  - adenylyl cyclase 8
OT  - breast cancer cells
OT  - migration
OT  - orai1alpha
OT  - store-operated calcium entry
COIS- The authors declare no conflicts of interest.
EDAT- 2019/10/28 06:00
MHDA- 2019/10/28 06:01
PMCR- 2019/10/23
CRDT- 2019/10/27 06:00
PHST- 2019/10/10 00:00 [received]
PHST- 2019/10/21 00:00 [accepted]
PHST- 2019/10/27 06:00 [entrez]
PHST- 2019/10/28 06:00 [pubmed]
PHST- 2019/10/28 06:01 [medline]
PHST- 2019/10/23 00:00 [pmc-release]
AID - cancers11111624 [pii]
AID - cancers-11-01624 [pii]
AID - 10.3390/cancers11111624 [doi]
PST - epublish
SO  - Cancers (Basel). 2019 Oct 23;11(11):1624. doi: 10.3390/cancers11111624.