受体型酪氨酸磷酸酶J抑制血管内皮细胞增殖及迁移

Inhibition effect of PTPRJ over-expression on proliferation and migration of vascular endothelial cells

目的分析受体型酪氨酸磷酸酶J(PTPRJ)过表达对血管内皮细胞增殖及迁移活性的影响及机制。方法使用PTPRJ质粒转染人脐静脉内皮细胞(HUVEC)细胞。使用Western blot、RT-PCR、免疫荧光检测HUVEC细胞中PTPRJ表达情况,Western blot检测ERK1/2激酶磷酸化情况,RTPCR检测血小板源性生长因子-B(PDGF-B)的表达变化,Brd U摄入法检测细胞增殖率,划痕法检测细胞迁移率变化。结果免疫荧光检测质粒转染后HUVEC细胞中PTPRJ表达定位于细胞膜;PTPRJ质粒转染后ERK1/2激酶磷酸化受抑制,PDGF-B mRNA表达减少;PTPRJ质粒转染组细胞增殖率较空质粒组下降,差异有统计学意义(P=0.004)。PTPRJ质粒转染组细胞迁移率较空质粒组下降,差异有统计学意义(P=0.002)。结论 PTPRJ通过下调PDGF-B表达、抑制ERK1/2激酶磷酸化,对血管内皮细胞增殖及迁移活性具有负性调控作用。

Objective To investigate the effect of PTPRJ over-expression on cell proliferation and migration of vascular endothelial cells.Methods HUVEC cells were transfected by PTPRJ plasmids.PTPRJ expression was detected by immunofluorescence,Western blot and RT-PCR.ERK1/2 phosphorylation was measured by Western blot.Expression of PDGF-B was detected by RT-PCR.Cell proliferation rate was measured by BrdU nuclear incorporation assay.Cell migration rate was detected by wound healing assay.Results Immunofluorescence showed that PTPRJ was mainly distributed in cell membrane after PTPRJ plasmids transfection.ERK1/2 phosphorylation was inhibited and PDGF-B mRNA was down-regulated by PTPRJ plasmids transfection.Cell proliferation rate decreased after PTPRJ-WT transfection compared with empty vector transfection （P =0.004）.Cell migration rate decreased after PTPRJ-WT transfection compared with empty vector transfection （P =0.002）.Conclusion PTPRJ has critical negative influence on cell proliferation and migration of HUVEC cells by reversing ERK1/2 activation and inhibiting PDGF-B expression.