title: Silencing of STIM1 attenuates hypoxia-induced PASMCs proliferation via inhibition of the SOC/Ca2+/NFAT pathway.
abstract: BACKGROUND
  Stromal interaction molecule 1 (STIM1) is a newly discovered Ca2+ sensor on the endoplasmic reticulum which is an indispensable part in the activation of store-operated Ca2+ channels (SOC). Recent studies demonstrate that SOC of pulmonary smooth muscle cells (PASMCs) were upregulated by chronic hypoxia which contribute to the enhanced pulmonary vasoconstriction and vascular remodeling. However, the exact role of STIM1 in the development of chronic hypoxic pulmonary hypertension(HPH) remains unclear.
  
  
  METHODS
  In this study we investigated the cellular distribution and expression of STIM1 by immunofluorescence, qRTPCR and Western blotting methods in Wistar rat distal intrapulmonary arteries under normal and chronic hypobaric hypoxic conditions. In vitro, Wistar rat PASMCs were isolated and cultured. PASMCs were transfected with siRNA targeting STIM1 gene by liposome. The expression of STIM1 protein was detected by Western blotting. [3H]-thymidine ([3H]-TdR) incorporation were performed to detect PASMCs proliferation. The cell cycle was analyzed by flow cytometry. The SOC-mediated Ca2+ influx was calculated by Ca2+ fluorescence imaging and the nuclear translocation of NFATc3 was determined by immunofluorescence and Western blot analysis of nuclear extracts.
  
  
  RESULTS
  We found that during the development of HPH and the initiation of vascular remodeling, the mRNA and protein expression levels of STIM1 significantly increased in the distal intrapulmonary arteries. Moderate hypoxia significantly promotes PASMCs proliferation and cell cycle progression. Silencing of STIM1 significantly decreased cellular proliferation and delayed the cell cycle progression induced by hypoxia. Silencing of STIM1 also significantly decreased SOC-mediated Ca2+ influx and inhibited the nuclear translocation of NFATc3 in hypoxic PASMCs.
  
  
  CONCLUSIONS
  Our findings suggest that chronic hypobaric hypoxia upregulates the expression of STIM1 in the distal intrapulmonary arteries which plays an important role in the hypoxia-induced PASMCs proliferation via SOC/Ca2+/NFAT pathway and may represent a novel therapeutic target for the prevention of hypoxia pulmonary hypertension.
authors.affiliation: Department of Pathophysiology and high altitude physiology, College of high altitude military medicine, Third Military Medical University, Chongqing, China.
authors.forename: Xianhua
authors.initials: X
authors.lastname: Hou
authors.forename: Jian
authors.initials: J
authors.lastname: Chen
authors.forename: Yongjun
authors.initials: Y
authors.lastname: Luo
authors.forename: Fuyu
authors.initials: F
authors.lastname: Liu
authors.forename: Gang
authors.initials: G
authors.lastname: Xu
authors.forename: Yuqi
authors.initials: Y
authors.lastname: Gao
mesh_terms: D000818:Animals; D015220:Calcium Channels / Q000096:biosynthesis* / Q000235:genetics; D020013:Calcium Signaling / Q000502:physiology*; D049109:Cell Proliferation; D002478:Cells, Cultured; D020868:Gene Silencing / Q000502:physiology; D000860:Hypoxia / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D008297:Male; D008562:Membrane Glycoproteins / Q000096:biosynthesis* / Q000235:genetics; D009131:Muscle, Smooth, Vascular / Q000378:metabolism / Q000473:pathology; D032389:Myocytes, Smooth Muscle / Q000378:metabolism* / Q000473:pathology; D050778:NFATC Transcription Factors / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics; D011651:Pulmonary Artery / Q000378:metabolism / Q000473:pathology; D051381:Rats; D017208:Rats, Wistar; D000071737:Stromal Interaction Molecule 1
chemical_list: D015220:Calcium Channels; D008562:Membrane Glycoproteins; D050778:NFATC Transcription Factors; C518857:Stim1 protein, rat; D000071737:Stromal Interaction Molecule 1