Division of Women's Health, Women's Health Academic Centre, King's College London, King's Health Partners London, UK.
Physiology and Pharmacology, Faculty of Medicine, University of Calgary Calgary, AB, Canada.
Front Physiol. 2014 May 6;5:169. doi: 10.3389/fphys.2014.00169. eCollection 2014.
Store-operated calcium (Ca(2+)) entry (SOCE) can be mediated by two novel proteins, STIM/Orai. We have previously demonstrated that members of the TRPC family, putative basal and store operated calcium entry channels, are present in human myometrium and regulated by labor associated stimuli IL-1β and mechanical stretch. Although STIM and Orai isoforms (1-3) have been reported in other smooth muscle cell types, there is little known about the expression or gestational regulation of STIM and Orai expression in human myometrium. Total RNA was isolated from lower segment human myometrial biopsies obtained at Cesarean section from women at the time of preterm no labor (PTNL), preterm labor (PTL), term non-labor (TNL), and term with labor (TL); primary cultured human uterine smooth muscle cells, and a human myometrial cell line (hTERT-HM). STIM1-2, and Orai1-3 mRNA expression was assessed by quantitative real-time PCR. All five genes were expressed in myometrial tissue and cultured cells. STIM1-2 and Orai2-3 expression was significantly lower in cultured cells compared tissue. This has implications with regard investigation of the contribution of these proteins in cultured cells. Orai2 was the most abundant Orai isoform in human myometrium. Expression of STIM1-2/Orai1-3 did not alter with the onset of labor. Orai1 mRNA expression in cultured cells was enhanced by IL-1β treatment. This novel report of STIM1-2 and Orai1-3 mRNA expression in pregnant human myometrium and Orai1 regulation by IL-1β indicates a potential role for these proteins in calcium signaling in human myometrium during pregnancy.
钙库操纵性钙(Ca(2+))内流(SOCE)可由两种新型蛋白,即 STIM/Orai 介导。我们先前的研究表明,TRPC 家族成员(假定的基础和钙库操纵性钙内流通道)存在于人子宫平滑肌中,并受与分娩相关的刺激物 IL-1β 和机械拉伸调节。虽然在其他平滑肌细胞类型中已经报道了 STIM 和 Orai 同工型(1-3),但关于 STIM 和 Orai 在人子宫平滑肌中的表达或妊娠调节知之甚少。我们从剖宫产时取自未临产(PTNL)、早产临产(PTL)、足月未临产(TNL)和足月临产(TL)的女性的下段子宫肌活检中分离出总 RNA;从原代培养的人子宫平滑肌细胞和人子宫平滑肌细胞系(hTERT-HM)中分离出总 RNA。通过定量实时 PCR 评估 STIM1-2 和 Orai1-3 的 mRNA 表达。所有五个基因在子宫组织和培养细胞中均有表达。与组织相比,培养细胞中 STIM1-2 和 Orai2-3 的表达显著降低。这对于研究这些蛋白质在培养细胞中的作用具有重要意义。Orai2 是人类子宫平滑肌中最丰富的 Orai 同工型。STIM1-2/Orai1-3 的表达在分娩开始时没有改变。IL-1β 处理可增强培养细胞中 Orai1 的 mRNA 表达。本研究首次报道了 STIM1-2 和 Orai1-3 在妊娠人子宫平滑肌中的 mRNA 表达以及 IL-1β 对 Orai1 的调节,表明这些蛋白在妊娠期间人子宫平滑肌钙信号转导中具有潜在作用。