Effect of down-regulation of FABP5 on radiation damage of human keratinocytes
10.3760/cma.j.cn112271-20220921-00381
- VernacularTitle: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
- Author:
Hongdan GUAN
1
;
Rong ZHENG
;
Bingjie GUAN
;
Yuping LIN
;
Bisi WANG
;
Benhua XU
;
Jianyuan SONG
Author Information
1. 福建医科大学附属协和医院放疗科 福建省肿瘤智能影像与精准放疗重点实验室 福建省消化、血液系统与乳腺恶性肿瘤放射与治疗临床医学研究中心，福州 350001
- Keywords:
Skin;
Fatty acid binding protein 5;
Radiosensitivity;
Radiation protection;
PI3K/AKT signaling pathway
- From:
Chinese Journal of Radiological Medicine and Protection
2023;43(1):8-14
- CountryChina
- Language:Chinese
-
Abstract:
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.
