| 管鸿丹,郑榕,官秉洁,林玉萍,王弼思,徐本华,宋建元.下调脂肪酸结合蛋白5对人皮肤角质形成细胞放射损伤的影响[J].中华放射医学与防护杂志,2023,43(1):8-14 |
| 下调脂肪酸结合蛋白5对人皮肤角质形成细胞放射损伤的影响 |
| Effect of down-regulation of FABP5 on radiation damage of human keratinocytes |
| 投稿时间：2022-09-21 |
| DOI：10.3760/cma.j.cn112271-20220921-00381 |
| 中文关键词: 皮肤 脂肪酸结合蛋白5 放射敏感性 辐射防护 PI3K/AKT信号通路 |
| 英文关键词:Skin Fatty acid binding protein 5 Radiosensitivity Radiation protection PI3K/AKT signaling pathway |
| 基金项目:福建省卫生健康中青年骨干人才培养项目(2019-ZQN-44) |
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| 中文摘要: |
| 目的 探究下调脂肪酸结合蛋白5(FABP5)对皮肤细胞放射损伤的影响及其相关机制。方法 构建下调FABP5的慢病毒载体,将慢病毒感染人皮肤角质形成细胞(HaCaT)并检测转染效率。将细胞分为空白对照组、下调FABP5组、单纯照射组、下调FABP5+照射组。予6 MV X射线照射后,CCK-8法测定细胞增殖活力,划痕实验检测细胞迁移,流式细胞术分析细胞凋亡,克隆形成实验检测放射敏感性,免疫印迹检测细胞中PARP1、γ-H2AX、AKT、p-AKT的蛋白表达水平。结果 成功构建下调FABP5的HaCaT细胞,从RNA水平(t=25.14,P＜0.05)和蛋白水平(t=20.06,P＜0.05)验证均达到下调FABP5的目的。下调FABP5组细胞增殖(t=3.55、5.88、3.18,P＜0.05)、迁移能力(t=15.44,P＜0.05)显著减弱,其放射增敏比为0.782。下调FABP5+照射组细胞凋亡率较单纯照射组显著减少[(9.82±1.45)%vs.(22.05±6.71)%,t=3.08,P＜0.05]。下调FABP5+照射组细胞的PARP1和γ-H2AX蛋白水平分别为0.04±0.04、0.11±0.06和0.26±0.11、0.22±0.07,均低于单纯照射组的0.21±0.10、0.52±0.22和0.57±0.06、0.43±0.02(t=2.83、3.07、4.50、5.33,P＜0.05),而p-AKT蛋白水平高于单纯照射组(t=-16.24~3.02,P＜0.05)。结论 下调FABP5抑制皮肤细胞增殖、迁移,增强放射抵抗性,减少辐射诱导皮肤细胞凋亡和DNA损伤。这可能是通过磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(AKT)信号通路来抑制皮肤细胞放射敏感性,为放射性皮肤损伤防护提供一种新思路。 |
| 英文摘要: |
| Objective To investigate the effects of down-regulation of FABP5 (fatty acid binding protein 5) on radiation damage of skin cells, and explore underlying mechanism.Methods A lentiviral vector with down-regulated FABP5 was constructed to infect human immortalized keratinocytes (HaCaT) cells, and the transfection efficiency was examined. The HaCaT cells were divided into blank control group, FABP5 down-regulation group (FABP5), radiation group (IR), and FABP5 down-regulation combined with radiation group (FABP5+IR). After 6 MV X-ray radiation, cell proliferation viability was measured by CCK-8 assay, cell migration was detected by scratch assay, apoptosis was analyzed by flow cytometry, radiosensitivity was evaluated by cloning formation assay, and the cellular protein expressions of PARP1, γ-H2AX, AKT and p-AKT were detected by Western blot.Results FABP5 was successfully knocked-down in both RNA level (t=25.14, P<0.05) and protein level (t=20.06, P<0.05). The down-regulation of FABP5 decreased the abilities of cells proliferation (t=3.55, 5.88, 3.18, P<0.05) and migration (t=15.44, P<0.05), but increased cell resistance to irradiation with a radiosensitization ratio of 0.782. The apoptosis rate of FABP5+IR group was significantly lower than IR group (22.05±6.71)% vs. (9.82±1.45)%, t=3.08, P<0.05. The protein levels of PARP1 and γ-H2AX in FABP5+IR group were also lower than those in the IR group 0.04±0.04, 0.11±0.06, 0.26±0.11, 0.22±0.07, 0.21±0.10, 0.52±0.22, 0.57±0.06, 0.43±0.02(t=2.83, 3.07, 4.50, 5.33, P<0.05), while the protein level of p-Akt in FABP5+IR group was higher than that in IR group (t=-16.24—3.02, P<0.05).Conclusions Down-regulation of FABP5 inhibited cell proliferation and migration, increased radioresistance, and reduced radiation-induced apoptosis and DNA damage of skin cells probably through PI3K/AKT signaling pathway. |
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