VEGFC/VEGFR3 Signaling Regulates Mouse Spermatogonial Cell Proliferation via the Activation of AKT/MAPK and Cyclin D1 Pathway and Mediates the Apoptosis by affecting Caspase 3/9 and Bcl-2
VEGFC/VEGFR3 Signaling Regulates Mouse Spermatogonial Cell Proliferation via the Activation of AKT/MAPK and Cyclin D1 Pathway and Mediates the Apoptosis by affecting Caspase 3/9 and Bcl-2
复制标题
VEGFC/VEGFR3 信号传导通过激活 AKT/MAPK 和 Cyclin D1 通路调节小鼠精原细胞增殖，并通过影响 Caspase 3/9 和 Bcl-2 介导细胞凋亡
DOI：
10.1080/15384101.2017.1407891
复制
发表时间：
2018-01-01
期刊：
影响因子：
4.3
通讯作者：
Li, Zheng
中科院分区：
文献类型：
--
作者：
Zhao, Liangyu;Zhu, Zijue;Li, Zheng
我们先前已经表明，Vegfc及其受体Vegfr 3的转录水平在精原细胞中很高，在精母细胞和精子细胞中极低。然而，VEGFC/VEGFR 3信号通路在精原细胞命运决定中的作用和机制尚不清楚。为此，在这里，我们探讨了VEGFC/VEGFR 3的作用和信号通路，通过使用来自永生化的小鼠精原细胞保留有丝分裂生殖细胞的标志物，即GC-1细胞。VEGFR 3在小鼠初级精原细胞和GC-1细胞中表达。VEGFC可促进GC-1细胞增殖和DNA合成，增强PI 3 K-AKT和MAPK的磷酸化，而AKT抑制剂LY 294002和MAPK抑制剂CI-1040可阻断VEGFC对GC-1细胞增殖的影响。此外，VEGFC增加c-fos和Egr 1的转录和cyclin D1，PCNA和Bcl-2的蛋白水平。相反，通过VEGFR 3敲低阻断VEGFC/VEGFR 3信号传导减少了AKT/MAPK的磷酸化，并降低了细胞周期蛋白D1和PCNA的水平。此外，VEGFR 3基因敲低不仅导致GC-1细胞凋亡增加，而且还导致Bcl-2表达减少，Caspase-3/9和PARP的裂解增加。综上所述，VEGFC/VEGFR 3信号通路通过AKT /MAPK和cyclin D1通路促进GC-1细胞增殖，通过Caspase-3/9、PARP和Bcl-2抑制细胞凋亡。因此，这项研究为哺乳动物精原细胞命运决定的分子机制提供了新的见解。
We have previously shown that the transcript levels of Vegfc and its receptor Vegfr3 were high in spermatogonia and extremely low in spermatocytes and spermatids. However, it remains unknown about the functions and the mechanisms of VEGFC/VEGFR3 signaling in regulating the fate determinations of spermatogonia. To this end, here we explored the role and signaling pathways of VEGFC/VEGFR3 by using a cell line derived from immortalized mouse spermatogonia retaining markers of mitotic germ cells, namely GC-1 cells. VEGFR3 was expressed in mouse primary spermatogonia and GC-1 cells. VEGFC stimulated the proliferation and DNA synthesis of GC-1 cells and enhanced the phosphorylation of PI3K-AKT and MAPK, whereas LY294002 (an inhibitor for AKT) and CI-1040 (an inhibitor for MAPK) blocked the effect of VEGFC on GC-1 cell proliferation. Furthermore, VEGFC increased the transcripts of c-fos and Egr1 and protein levels of cyclin D1, PCNA and Bcl-2. Conversely, the blocking of VEGFC/VEGFR3 signaling by VEGFR3 knockdown reduced the phosphorylation of AKT/MAPK and decreased the levels of cyclin D1 and PCNA. Additionally, VEGFR3 knockdown not only resulted in more apoptosis of GC-1 cells but also led to a decrease of Bcl-2 and promoted the cleavage of Caspase-3/9 and PARP. Collectively, these data suggested that VEGFC/VEGFR3 signaling promotes the proliferation of GC-1 cells via the AKT /MAPK and cyclin D1 pathway and it inhibits the cell apoptosis through Caspase-3/9, PARP and Bcl-2. Thus, this study sheds a novel insight to the molecular mechanisms underlying the fate decisions of mammalian spermatogonia.