Effect of graphene oxide in promoting HaCaT cell migration
in vitro and possible mechanisms- Journal of Third Military Medical University Vol. 40, Issue 10, Pages: 889-897(2018)
- 作者机构：
1.400038 重庆，陆军军医大学(第三军医大学)第一附属医院全军烧伤研究所，创伤、烧伤与复合伤国家重点实验室
2.610064 成都，四川大学高分子研究所，高分子材料工程国家重点实验室
3.MB R3T 2N2加拿大 温哥华，曼尼托巴大学和曼尼托巴儿童健康研究所，机械工程、生物化学和医学遗传学系
- 作者简介：
- 基金信息：the Key Program of National Natural Science Foundation of China(81630055);the Major Technology Innovation of Southwest Hospital Program(SWH2016ZDCX2014)
DOI：10.16016/j.1000-5404.201712234
CLC： R318.08;R322.99;R329.28Received：25 December 2017，
Revised：2018-02-07，
Published：30 May 2018
- 稿件说明：
移动端阅览
Tao YANG, Daijun ZHOU, Song WANG, et al. Effect of graphene oxide in promoting HaCaT cell migration
in vitro and possible mechanisms[J]. Journal of Third Military Medical University, 2018, 40(10): 889-897.Tao YANG, Daijun ZHOU, Song WANG, et al. Effect of graphene oxide in promoting HaCaT cell migration
in vitro and possible mechanisms[J]. Journal of Third Military Medical University, 2018, 40(10): 889-897. DOI： 10.16016/j.1000-5404.201712234.
摘要
目的
2
观察氧化石墨烯(grapheneoxide
GO)对人表皮细胞株HaCaT细胞迁移的影响，初步探讨其可能机制。
方法
2
将HaCaT细胞分别应用GO终浓度为0、0.001、0.01、0.1、1.0、10.0 μg/mL的RPMI1640细胞培养液培养，在培养的不同时相点，应用CCK-8法检测细胞活性，蛋白印迹法检测PCNA表达。采用划痕实验检测细胞迁移情况，激光共聚焦显微镜观察细胞骨架改变，蛋白质印迹法检测细胞F-actin、Vinculin的表达。
结果
2
(1) 低浓度GO对HaCaT细胞增殖无影响，当GO浓度高于1.0 μg/mL时则具有明显的毒性作用。10.0 μg/mL GO可明显抑制细胞PCNA表达(
P
<
0.05)。(2)划痕实验结果显示：低浓度GO可促进HaCaT细胞迁移，其中以0.01 μg/mL浓度效果最明显，在划痕后24 h，该组细胞迁移面积比对照组增加35.33% (
P
<
0.01)。(3)激光共聚焦显微镜观察发现，0.01 μg/mL GO刺激后，细胞纤毛明显增多，胞内应力性纤维增粗。而其1.0 μg/mL组细胞变小、变圆。(4)蛋白印迹法检测结果显示：0.01 μg/mL GO刺激后F-actin表达增加(
P
<
0.05)，而1.0 μg/mL作用后F-actin表达却明显低于对照组。GO能促进Vinculin表达，但仅当GO浓度高于0.1 μg/mL后差异才有统计学意义。
结论
2
特定浓度的GO可促进HaCaT细胞迁移，可能与其影响细胞骨架及F-actin、Vinculin表达有关。
Abstract
Objective
2
To investigate the effect of graphene oxide (GO) on the migration of human epidermal cell line (HaCaT cells) and explore the possible mechanisms.
Methods
2
HaCaT cells were cultured in RPMI1640 media containing different concentrations of GO for 24~72 h
and the changes in cell viability and proliferation were examined using the cell counting kit-8 (CCK-8) assay. The changes in cell migration were assessed in a cell scratch model. The cytoskeletons of the treated cells were observed using confocal laser scanning microscope (CLSM)
and the expression levels of proliferating cell nuclear antigen (PCNA)
F-actin and vinculin after GO treatment were detected using Western blotting.
Results
2
Compared with the control cells
the cells exposed to low concentrations of GO for 24 h exhibited no significant changes in cell viability
while GO at concentrations above 1.0 μg/mL caused obvious cytotoxicity (
P
<
0.01)
and treatment with 10.0 μg/mL GO for 24 h resulted in significantly decreased PCNA expression in the cells (
P
<
0.05). Scratch wound assay showed that GO at low concentrations was capable of promoting the migr
ation of HaCaT cells
and the effect was the strongest at 0.01 μg/mL
which significantly increased the cell migration rate by 35.33% at 24 h (
P
<
0.01); GO at 1.0 μg/mL obviously inhibited the cell migration (
P
<
0.01). CLSM demonstrated an obviously increased number of cell cilia and thickened cell stress fibers in cells exposed to 0.01 μg/mL GO. Incubation of the cells with 0.01 μg/mL GO significantly increased the expression level of F-actin
while 1.0 μg/mL GO significantly suppressed F-actin expression (
P
<
0.05). GO also enhanced the expression of vinculin in the cells
and the changes were statistically significant when GO concentration was above 0.1 μg/mL.
Conclusion
2
GO within a defined concentration range can significantly promote the migration of HaCaT cells
the mechanism of which may involve the regulation of F-actin and vinculin expressions and cytoskeleton rearrangement.
关键词
Keywords
references
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