Functional TRPV2 and TRPV4 channels in human cardiac c-kit(+) progenitor cells.
Functional TRPV2 and TRPV4 channels in human cardiac c-kit(+) progenitor cells.
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DOI：
10.1111/jcmm.12800
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发表时间：
2016-06
影响因子：
5.3
通讯作者：
Li GR
中科院分区：
文献类型：
--
作者：
Che H;Xiao GS;Sun HY;Wang Y;Li GR
人类心脏c-kit+祖细胞的细胞生理学和生物学特性尚未得到广泛的表征，仍是一个活跃的研究领域。本研究探讨瞬时受体电位香草酸(TRPV)的功能表达及其在调节人心脏c-kit+祖细胞增殖和迁移中的可能作用。我们发现编码TRPV2和TRPV4通道的基因及其蛋白在人c-kit+心脏干细胞中显著表达。TRPV2的激活剂丙磺舒可引起细胞内钙离子浓度的增加，这种作用可被TRPV2阻滞剂Ru红减弱或消除。TRPV4通道激活剂4α-佛波酯引起的钙振荡可被TRPV4阻断剂RN-1734抑制。在TRPV2shRNA或TRPV4shRNA感染的细胞中，丙磺舒或4α-佛波酯对细胞内钙离子浓度的变化有明显的抑制作用。沉默TRPV2，而不是TRPV4，通过将细胞停滞在细胞周期的G0/G1边界而显著降低细胞的增殖。沉默TRPV2或TRPV4可减少细胞迁移。Western印迹显示，沉默TRPV2可降低细胞周期蛋白D1、细胞周期蛋白E、pERK1/2和PACK的表达，而沉默TRPV4仅能降低PACT的表达。我们的结果首次证明在人心脏c-kit+祖细胞中大量表达有功能的TRPV2和TRPV4通道。TRPV2通道，而不是TRPV4通道，参与调节细胞周期进程；此外，TRPV2和TRPV4都参与人类心脏c-kit+祖细胞的迁移。
The cellular physiology and biology of human cardiac c‐kit+ progenitor cells has not been extensively characterized and remains an area of active research. This study investigates the functional expression of transient receptor potential vanilloid (TRPV) and possible roles for this ion channel in regulating proliferation and migration of human cardiac c‐kit+ progenitor cells. We found that genes coding for TRPV2 and TRPV4 channels and their proteins are significantly expressed in human c‐kit+ cardiac stem cells. Probenecid, an activator of TRPV2, induced an increase in intracellular Ca2+ (Ca2+ i), an effect that may be attenuated or abolished by the TRPV2 blocker ruthenium red. The TRPV4 channel activator 4α‐phorbol 12‐13‐dicaprinate induced Ca2+ i oscillations, which can be inhibited by the TRPV4 blocker RN‐1734. The alteration of Ca2+ i by probenecid or 4α‐phorbol 12‐13‐dicprinate was dramatically inhibited in cells infected with TRPV2 short hairpin RNA (shRNA) or TRPV4 shRNA. Silencing TRPV2, but not TRPV4, significantly reduced cell proliferation by arresting cells at the G0/G1 boundary of the cell cycle. Cell migration was reduced by silencing TRPV2 or TRPV4. Western blot revealed that silencing TRPV2 decreased expression of cyclin D1, cyclin E, pERK1/2 and pAkt, whereas silencing TRPV4 only reduced pAkt expression. Our results demonstrate for the first time that functional TRPV2 and TRPV4 channels are abundantly expressed in human cardiac c‐kit+ progenitor cells. TRPV2 channels, but not TRPV4 channels, participate in regulating cell cycle progression; moreover, both TRPV2 and TRPV4 are involved in migration of human cardiac c‐kit+ progenitor cells.