The Effect Of Calcium Channel On Cell Proliferation, Migration And Invasion

BackgroundStore-operated Ca2+entry (SOCE) is a common mechanism to regulate Ca2+influx intocells and participates in many biological processes including cell proliferation. STIM1protein and Orai1protein are important components of SOCE. Glomerular mesangialcells (GMCs) involve in the regulation of the glomerular filtration rate. From a clinicalpoint of view, many physiological functions will change with aging.ObjectiveTo investigate the role of SOCE in mesangial cell proliferation in aged rats and themechanism.Methods1SOCE in GMCs of young and aged ratsSOCE was activated by Ca2+release induced by physiological agonists inprimary-cultured young (3months old) and aged (22months old) rat GMCs. UsingLeica TCS SP5confocal system to detect changes of intracellular Ca2+concentration.2Expression levels of STIM1and Orai1in glomeruli from young and aged ratsWestern Blot and immunofluorescence were used to detect the expression levels ofSTIM1and Orai1in glomeruli from young and aged rats. 3The roles of STIM1and Orai1in rat GMCs SOCESTIM1siRNA or Orai1siRNA were transfected into the primary-cultured young ratGMCs, and the Leica TCS SP5confocal system was used to detect changes ofintracellular Ca2+concentration. Using immunofluorescence to detect STIM1and Orai1puncta formation induced by Ca2+store depletion in primary-cultured young and agedrat GMCs.4The proliferation capability of aged rat GMCs and the mechanism.Bromodeoxyuridine assay and MTT assay were used to detect the proliferationcapability of young and aged rat GMCs, and primary-cultured young rat GMCstransfected with STIM1siRNA or Orai1siRNA.Results1SOCE induced by physiological agonists was significantly attenuated in aged ratGMCs compared with that in younger rats.2The expression of both STIM1and Orai1proteins was significantly reduced inaged rat glomeruli compared with that in younger rats.3SOCE was significantly attenuated in primary-cultured young rat GMCs transfectedwith STIM1siRNA or Orai1siRNA.4Ca2+store depletion significantly induced STIM1and Orai1puncta formation inyoung rats GMCs.5STIM1and Orai1puncta formation was significantly weakened compared withthat in younger rats. 6The proliferation of aged rat GMCs was significantly attenuated compared with thatin younger rats.7The proliferation of primary-cultured young rat GMCs transfected with STIM1siRNA or Orai1siRNA was significantly attenuated.ConclusionThe attenuation of GMCs proliferation can be due to the decreased SOCE partiallycaused by reduced expression of STIM1and Orai1.