基础医学与临床 ›› 2019, Vol. 39 ›› Issue (9): 1243-1247.

• 研究论文 • 上一篇    下一篇

PODXL参与瘢痕疙瘩成纤维细胞的过度增殖

郭亚东,吴红娟,孙要文   

  1. 陕西省人民医院
  • 收稿日期:2018-06-20 修回日期:2018-12-05 出版日期:2019-09-05 发布日期:2019-09-06
  • 通讯作者: 孙要文 E-mail:sunyaowen@yeah.net
  • 基金资助:
    陕西省自然基金课题

PODXL participates in the excessive proliferation of keloid fibroblasts

  • Received:2018-06-20 Revised:2018-12-05 Online:2019-09-05 Published:2019-09-06
  • Contact: sun yaowen E-mail:sunyaowen@yeah.net

摘要: 目的 探讨Podocalyxin类蛋白(PODXL)基因对瘢痕疙瘩成纤维细胞增殖、凋亡的影响。 方法 反转录-荧光定量PCR(RT-qPCR)检测正常皮肤成纤维细胞(hDF)和瘢痕疙瘩成纤维细胞(hKF)中PODXL mRNA的表达量;用重组PODXL基因的shRNA慢病毒表达载体感染瘢痕疙瘩成纤维细胞下调PODXL基因的表达量, EdU法检测细胞增殖,流式细胞仪检测细胞的周期和凋亡率,蛋白质印迹法(Western blot)检测细胞中磷脂酰肌醇-3激酶(PI3K)、丝苏氨酸蛋白激酶(AKT)总蛋白和磷酸化蛋白。 结果 与正常皮肤成纤维细胞相比,PODXL mRNA在瘢痕疙瘩成纤维细胞中的表达量显著增加(P<0.05);下调PODXL基因表达显著抑制瘢痕疙瘩成纤维细胞增殖(P<0.05),细胞周期阻滞于S期,促进细胞的凋亡(P<0.05),下降PI3K和AKT磷酸化蛋白水平(P<0.05)。 结论 PODXL在瘢痕疙瘩成纤维细胞中的表达量显著增加;下调PODXL基因表达可能通过抑制PI3K/AKT信号通路激活,从而将细胞周期阻滞在S期,抑制细胞的增殖和诱导凋亡。

关键词: 瘢痕疙瘩, 成纤维细胞, PODXL, PI3K/AKT

Abstract: Objective To investigate the effect of Podocalyxin protein (PODXL) on the proliferation and apoptosis of keloid fibroblasts. Methods Reverse transcription quantitative PCR (RT-qPCR) was used to detect the expression of PODXL mRNA in normal skin fibroblasts and keloid fibroblasts. The expression of PODXL gene in keloid fibroblasts was inhibited by recombinant PODXL gene shRNA lentiviral expression vector infection. The proliferation of cells was measured by EdU assay. The cell cycle and apoptosis rate were analyzed by flow cytometry. The level of phosphatidylinositol 3-kinase (PI3K), tyrosine kinase (AKT) total protein and phosphorylated protein in cells were detected by Western Blot. Results Compared with normal skin fibroblasts, the expression of PODXL mRNA in keloid fibroblasts was significantly increased(P<0.05). Down-regulation of PODXL gene expression significantly inhibited the proliferation of keloid fibroblasts(P<0.05), arrested the cell cycle in S phase and promoted apoptosis(P<0.05), decreased PI3K, AKT phosphorylation protein levels(P<0.05). Conclusion The expression of PODXL in keloid fibroblasts is significantly increased. Down-regulation of PODXL gene expression may arrest cell cycle in S phase, inhibit proliferation and induce apoptosis by inhibit the activation of PI3K/AKT signaling pathway.

Key words: Keloid, fibroblasts, PODXL, PI3K/AKT