Abstract
Background: Orai1/CRACM1 is a principal component of the store-operated calcium channels. Store-operated calcium influx is highly correlated with inflammatory reactions, immunological regulation, and cell proliferation. Epidermal growth factor (EGF), which plays an important role in the regulation of cell proliferation, can activate store-operated calcium channels. However, the consequences of Orai1/CRACM1 overexpression in EGF-mediated lung cancer cells growth are not known. Methods: To investigate the role of Orai1/CRACM1 in EGF-mediated lung cancer cell proliferation, Orai1/CRACM1 plasmids were transfected into cells by lipofection. A cell proliferation assay, immunofluorescence staining, flow cytometry, and real-time polymerase chain reaction were employed to monitor cell proliferation. The calcium influx signals were investigated using a fluorescent-based calcium assay. Results: Transfection of Orai1/CRACM1 plasmids resulted in the inhibition of EGF-mediated cell proliferation. ERK1/2 and Akt phosphorylation were inhibited by Orai1/CRACM1 overexpression. Expression of the cell cycle modulator p21 was induced in the Orai1/CRACM1-overexpressing cells, whereas the expression of cyclin D3 was reduced. Flow cytometry revealed that overexpression of Orai1/CRACM1 resulted in G0/G1 cell cycle arrest. Importantly, Orai1/CRACM1 overexpression significantly attenuated EGF-mediated store-operated calcium influx. In addition, application of 2-APB, a store-operated calcium channel inhibitor, resulted in the inhibition of EGF-mediated cancer cell proliferation. Conclusions: We conclude that Orai1/CRACM1 overexpression attenuates store-operated Ca 2+ influx that in turn blocks EGF-mediated proliferative signaling and drives cell cycle arrest.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1278-1284 |
| Number of pages | 7 |
| Journal | Biochimica et Biophysica Acta - General Subjects |
| Volume | 1810 |
| Issue number | 12 |
| DOIs | |
| State | Published - Dec 1 2011 |
Fingerprint
All Science Journal Classification (ASJC) codes
- Biophysics
- Biochemistry
- Molecular Biology
Cite this
}
Orai1/CRACM1 overexpression suppresses cell proliferation via attenuation of the store-operated calcium influx-mediated signalling pathway in A549 lung cancer cells. / Hou, Ming Feng; Kuo, Ho Chang; Li, Jih Heng; Wang, Yu Shiuan; Chang, Chen Chia; Chen, Ku Chung; Chen, Wei Chiao; Chiu, Chien Chih; Yang, Shengyu; Chang, Wei Chiao.
In: Biochimica et Biophysica Acta - General Subjects, Vol. 1810, No. 12, 01.12.2011, p. 1278-1284.Research output: Contribution to journal › Article
TY - JOUR
T1 - Orai1/CRACM1 overexpression suppresses cell proliferation via attenuation of the store-operated calcium influx-mediated signalling pathway in A549 lung cancer cells
AU - Hou, Ming Feng
AU - Kuo, Ho Chang
AU - Li, Jih Heng
AU - Wang, Yu Shiuan
AU - Chang, Chen Chia
AU - Chen, Ku Chung
AU - Chen, Wei Chiao
AU - Chiu, Chien Chih
AU - Yang, Shengyu
AU - Chang, Wei Chiao
PY - 2011/12/1
Y1 - 2011/12/1
N2 - Background: Orai1/CRACM1 is a principal component of the store-operated calcium channels. Store-operated calcium influx is highly correlated with inflammatory reactions, immunological regulation, and cell proliferation. Epidermal growth factor (EGF), which plays an important role in the regulation of cell proliferation, can activate store-operated calcium channels. However, the consequences of Orai1/CRACM1 overexpression in EGF-mediated lung cancer cells growth are not known. Methods: To investigate the role of Orai1/CRACM1 in EGF-mediated lung cancer cell proliferation, Orai1/CRACM1 plasmids were transfected into cells by lipofection. A cell proliferation assay, immunofluorescence staining, flow cytometry, and real-time polymerase chain reaction were employed to monitor cell proliferation. The calcium influx signals were investigated using a fluorescent-based calcium assay. Results: Transfection of Orai1/CRACM1 plasmids resulted in the inhibition of EGF-mediated cell proliferation. ERK1/2 and Akt phosphorylation were inhibited by Orai1/CRACM1 overexpression. Expression of the cell cycle modulator p21 was induced in the Orai1/CRACM1-overexpressing cells, whereas the expression of cyclin D3 was reduced. Flow cytometry revealed that overexpression of Orai1/CRACM1 resulted in G0/G1 cell cycle arrest. Importantly, Orai1/CRACM1 overexpression significantly attenuated EGF-mediated store-operated calcium influx. In addition, application of 2-APB, a store-operated calcium channel inhibitor, resulted in the inhibition of EGF-mediated cancer cell proliferation. Conclusions: We conclude that Orai1/CRACM1 overexpression attenuates store-operated Ca 2+ influx that in turn blocks EGF-mediated proliferative signaling and drives cell cycle arrest.
AB - Background: Orai1/CRACM1 is a principal component of the store-operated calcium channels. Store-operated calcium influx is highly correlated with inflammatory reactions, immunological regulation, and cell proliferation. Epidermal growth factor (EGF), which plays an important role in the regulation of cell proliferation, can activate store-operated calcium channels. However, the consequences of Orai1/CRACM1 overexpression in EGF-mediated lung cancer cells growth are not known. Methods: To investigate the role of Orai1/CRACM1 in EGF-mediated lung cancer cell proliferation, Orai1/CRACM1 plasmids were transfected into cells by lipofection. A cell proliferation assay, immunofluorescence staining, flow cytometry, and real-time polymerase chain reaction were employed to monitor cell proliferation. The calcium influx signals were investigated using a fluorescent-based calcium assay. Results: Transfection of Orai1/CRACM1 plasmids resulted in the inhibition of EGF-mediated cell proliferation. ERK1/2 and Akt phosphorylation were inhibited by Orai1/CRACM1 overexpression. Expression of the cell cycle modulator p21 was induced in the Orai1/CRACM1-overexpressing cells, whereas the expression of cyclin D3 was reduced. Flow cytometry revealed that overexpression of Orai1/CRACM1 resulted in G0/G1 cell cycle arrest. Importantly, Orai1/CRACM1 overexpression significantly attenuated EGF-mediated store-operated calcium influx. In addition, application of 2-APB, a store-operated calcium channel inhibitor, resulted in the inhibition of EGF-mediated cancer cell proliferation. Conclusions: We conclude that Orai1/CRACM1 overexpression attenuates store-operated Ca 2+ influx that in turn blocks EGF-mediated proliferative signaling and drives cell cycle arrest.
UR - http://www.scopus.com/inward/record.url?scp=80855148154&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=80855148154&partnerID=8YFLogxK
U2 - 10.1016/j.bbagen.2011.07.001
DO - 10.1016/j.bbagen.2011.07.001
M3 - Article
C2 - 21782006
AN - SCOPUS:80855148154
VL - 1810
SP - 1278
EP - 1284
JO - Biochimica et Biophysica Acta - General Subjects
JF - Biochimica et Biophysica Acta - General Subjects
SN - 0006-3002
IS - 12
ER -