果糖-1,6-二磷酸醛缩酶 A 通过诱导片状伪足形成参与 HaCaT 细胞迁移。

Fructose-1,6-bisphosphate aldolase A is involved in HaCaT cell migration by inducing lamellipodia formation.

机构信息

Graduate School of Biological Science, Hiroshima University, Higashihiroshima, Japan.

出版信息

J Dermatol Sci. 2010 May;58(2):123-9. doi: 10.1016/j.jdermsci.2010.02.012. Epub 2010 Mar 6.

DOI:10.1016/j.jdermsci.2010.02.012

PMID:20362419

Abstract

BACKGROUND

No previous report has investigated the involvement of glycolytic enzymes in keratinocyte migration. Fructose-1,6-bisphosphate aldolase A (ALDOA) is a glycolytic enzyme bound to the cytoskeleton by certain growth factors, which are known to enhance keratinocyte migration. We postulated that ALDOA is involved in keratinocyte migration.

OBJECTIVE

To investigate the possible role of ALDOA in keratinocyte migration.

METHODS

The localization of endogenous ALDOA and the actin cytoskeleton was observed by laser scanning confocal microscopy in HaCaT cells. The effects of ALDOA on lamellipodia formation and migration were evaluated using ALDOA siRNA-transfected cells. In addition, the involvement of epidermal growth factor (EGF) in ALDOA-induced events was investigated.

RESULTS

Strong ALDOA expression was observed along the ruffling membrane and lamellipodia, and it was colocalized with the actin cytoskeleton in lamellipodia. In a scratch wound assay, the wound recovery area was significantly decreased on transfection with ALDOA siRNA. The rate of lamellipodia-forming cells also decreased. On stimulation with EGF, the wound recovery area and ALDOA and its mRNA levels increased. On the other hand, ALDOA siRNA transfection suppressed EGF-enhanced migration.

CONCLUSION

We concluded that ALDOA is involved in keratinocyte migration following the induction of lamellipodia formation, and ALDOA-related migration is enhanced by EGF.

摘要

背景

目前尚无研究报道糖酵解酶在角质形成细胞迁移中的作用。果糖-1,6-二磷酸醛缩酶 A（ALDOA）是一种糖酵解酶，通过某些生长因子与细胞骨架结合，这些生长因子已知可增强角质形成细胞的迁移。我们假设 ALDOA 参与了角质形成细胞的迁移。

目的

研究 ALDOA 在角质形成细胞迁移中的可能作用。

方法

使用激光共聚焦显微镜观察 HaCaT 细胞中内源性 ALDOA 和肌动蛋白细胞骨架的定位。使用 ALDOA siRNA 转染细胞评估 ALDOA 对片状伪足形成和迁移的影响。此外，还研究了表皮生长因子（EGF）在 ALDOA 诱导事件中的作用。

结果

在皱襞膜和片状伪足中观察到强烈的 ALDOA 表达，并且在片状伪足中与肌动蛋白细胞骨架共定位。在划痕实验中，ALDOA siRNA 转染后伤口恢复面积明显减小。形成片状伪足的细胞比例也降低了。EGF 刺激后，伤口恢复面积和 ALDOA 及其 mRNA 水平增加。另一方面，ALDOA siRNA 转染抑制了 EGF 增强的迁移。