Zhonghua Xin Xue Guan Bing Za Zhi. 2011 Jul;39(7):649-53.
[Impact of stromal interaction molecule 1 silencing on cell cycle of endothelial progenitor cells].
Kuang, C. Y., Huang, L., Yu, Y., Deng, M. Y., Wang, K., Qian, D. H.,
["Cardiovascular Research Institute of Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China."]
["Cardiovascular Research Institute of Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China."]
OBJECTIVE: To investigate the effect of stromal interaction molecule 1 (STIM1) silencing on EPCs cell cycle. METHODS: Rat bone marrow derived endothelial progenitor cells (EPCs) were isolated and cultured in L-DMEM with 20% FBS. Ad-si/rSTIM1 and Ad-hSTIM1 were then transfected into EPCs and the expression of STIM1 mRNA was detected by RT-PCR. The cell cycle was determined using flow cytometry analysis and intracellular free Ca2+ was measured using LSCM. Co-immunoprecipitation was performed to examine the interaction between STIM1 and TRPC1. Protein levels of inositol 1, 4, 5-trisphosphate were analyzed with ELISA assay. RESULTS: Forty-eight hours after transfection, the expression of STIM1 mRNA was significantly downregulated (0.37 +/- 0.02 vs. 1.00 +/- 0.02, P < 0.05) and intracellular free Ca2+ level was significantly reduced (34.07 +/- 4.10 vs. 86.51 +/- 14.12, P < 0.05) in Ad-si/rSTIM1 group compared with control group. The cell cycle was arrested at G1 phase [(90.91 +/- 1.10)% vs. (77.10 +/- 0.56)%, P < 0.05] and the store-operated channel entry was strikingly inhibited in EPCs after treatment with Ad-si/rSTIM1. However, cotransfection of Ad-hSTIM1 with Ad-si/rSTIM1 significantly reversed these responses. Interestingly, co-immunoprecipitation study showed that STIM1 co-precipitated with TRPC1, and IP3 levels measured by ELISA were similar among three groups (P > 0.05). CONCLUSION: siRNA-mediated knockdown of STIM1 inhibited EPCs proliferation by reducing intracellular free Ca2+ through TRPC1-SOC signaling pathway.
PMID: 22088247
 Screening
Validation: In vitro validation
Validation: In vivo validation
Characterization
Functional consequence
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Validation: In vivo validation
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| Assay with endogenous proteins | Assay with overexpressed proteins | Reference | ||||||||
| Cell or tissue | Cell or tissue | TRP channel construct | Interactor construct | |||||||
| TRP channel | Interactor | Method | Species | Region | Species | Region | ||||
| TRPC2 |
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Epo-R | Co-immunoprecipitation | CHO-S | Mouse | Full-length | Mouse | Full-length | 22088247 | |
| TRPC2 |
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IP3R2 | Co-immunoprecipitation | CHO-S | Mouse | Full-length | Rat | Full-length | 22088247 | |
| TRPC2 |
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PLCƣ1 | Co-immunoprecipitation | CHO-S | Mouse | Full-length | Rat | Full-length | 22088247 | |
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click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
:
click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)