Deletion of the PDGFR-β gene affects key fibroblast functions important for wound healing
Deletion of the PDGFR-β gene affects key fibroblast functions important for wound healing
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DOI：
10.1074/jbc.m413081200
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发表时间：
2005-03-11
影响因子：
4.8
通讯作者：
Sasahara, M
中科院分区：
文献类型：
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作者：
Gao, ZY;Sasaoka, T;Sasahara, M
这项研究提供了新的观点的独特方面的血小板衍生生长因子β受体（PDGFR-β）的信号转导和生物反应，通过建立一个突变的小鼠品系，其中两个loxP序列插入到PDGFR-β基因组序列的内含子。从突变小鼠中分离皮肤成纤维细胞并在体外转染Cre重组酶诱导PDGFR-β基因缺失（PDGFR-β（Delta/Delta））。PDGFR-β蛋白的最终消耗显著减弱了血小板衍生生长因子（PDGF）-BB诱导的细胞迁移、增殖，并保护了培养的PDGFR-β（Delta/Delta）真皮成纤维细胞免于H2 O2诱导的细胞凋亡。PDGF-AA和胎牛血清具有促有丝分裂和抗凋亡作用，但不能诱导PDGFR-β（Delta/Delta）成纤维细胞迁移。关于PDGF信号传导，PDGF-BB诱导的Akt、ERK 1/2和JNK磷酸化在PDGFR-β（Delta/Delta）成纤维细胞中减少，但不影响p38，但PDGF-AA诱导的信号传导没有改变。过表达的磷脂磷酸酶，SHIP 2和/或PTEN，抑制PDGF-BB诱导的磷酸化Akt和ERK 1/2的PDGFR-β（三角洲/三角洲）成纤维细胞，但不影响JNK和p38。这些结果表明，PDGFR-β（Delta/Delta）真皮成纤维细胞中不同PDGFR-β信号通路的破坏损害了它们的增殖和存活，但完全抑制了迁移反应，并且PDGF-BB诱导的Akt和ERK 1/2磷酸化可能由PDGFR-α介导，至少部分由脂质磷酸酶SHIP 2和/或PTEN调节。因此，皮肤成纤维细胞上的PDGFR-β功能似乎在PDGF-BB对皮肤伤口愈合的作用中是关键的，并且在配体诱导的生物反应和细胞信号传导的潜在性质中与PDGFR-β的功能明显不同。
This study provides new perspectives of the unique aspects of platelet-derived growth factor beta-receptor (PDGFR-beta) signaling and biological responses through the establishment of a mutant mouse strain in which two loxP sequences were inserted into the introns of PDGFR-beta genome sequences. Isolation of skin fibroblasts from the mutant mice and Cre recombinase transfection in vitro induced PDGFR-beta gene deletion ( PDGFR-beta(Delta/Delta)). The resultant depletion of the PDGFR-beta protein significantly attenuated platelet-derived growth factor ( PDGF)-BB-induced cell migration, proliferation, and protection from H2O2-induced apoptosis of the cultured PDGFR-beta(Delta/Delta) dermal fibroblasts. PDGF-AA and fetal bovine serum were mitogenic and antiapoptotic but were unable to induce the migration in PDGFR-beta(Delta/Delta) fibroblasts. Concerning the PDGF signaling, PDGF-BB-induced phosphorylation of Akt, ERK1/2, and JNK, but not p38, decreased in PDGFR-beta(Delta/Delta) fibroblasts, but PDGF-AA-induced signaling was not altered. Overexpression of the phospholipid phosphatases, SHIP2 and/or PTEN, inhibited PDGF-BB-induced phosphorylation of Akt and ERK1/2 in PDGFR-beta(Delta/Delta) fibroblasts but did not affect that of JNK and p38. These results indicate that disruption of distinct PDGFR-beta signaling pathways in PDGFR-beta(Delta/Delta) dermal fibroblasts impaired their proliferation and survival, but completely inhibits migratory response, and that PDGF-BB-induced phosphorylation of Akt and ERK1/2 possibly mediated by PDGFR-alpha is regulated, at least in part, by the lipid phosphatases SHIP2 and/or PTEN. Thus, the PDGFR-beta function on dermal fibroblasts appears to be critical in PDGF-BB action for skin wound healing and is clearly distinctive from that of PDGFR-beta in the ligand-induced biological responses and the underlying properties of cellular signaling.