Regulatory effects of bio-intensity electric field on microtubule acetylation in human epidermal cell line HaCaT

Wu Yating; Zhang Ze; Ji Ran; Zhang Shuhao; Wang Wenping; Wu Chao; Zhang Jiaping; Jiang Xupin; Zhang Hengshu

doi:10.3760/cma.j.cn501120-20211105-00377

Volume 38 Issue 11

Nov. 2022

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Article Navigation > CHINESE JOURNAL OF BURNS AND WOUNDS > 2022 > 38(11): 1066-1072

Wu YT,Zhang Z,Ji R,et al.Regulatory effects of bio-intensity electric field on microtubule acetylation in human epidermal cell line HaCaT[J].Chin J Burns Wounds,2022,38(11):1066-1072.DOI: 10.3760/cma.j.cn501120-20211105-00377.

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Regulatory effects of bio-intensity electric field on microtubule acetylation in human epidermal cell line HaCaT

doi: 10.3760/cma.j.cn501120-20211105-00377

1.
Department of Plastic and Burn Surgery, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China
2.
Department of Plastic Surgery, the First Affiliated Hospital of Army Medical University (the Third Military Medical University), Chongqing 400038, China

Funds:

Gerenal Program of National Natural Science Foundation of China 82072172

Youth Found of National Natural Science Foundation of China 81601683

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Corresponding author: Jiang Xupin, Email: jiangxupin@126.com; Zhang Hengshu, Email: 1390527030@qq.com
Received Date: 2021-11-05

Abstract

Abstract

Objective To investigate the regulatory effects of bio-intensity electric field on directional migration and microtubule acetylation in human epidermal cell line HaCaT, aiming to provide molecular theoretical basis for the clinical treatment of wound repair. Methods The experimental research methods were used. HaCaT cells were collected and divided into simulated electric field group (n=54) placed in the electric field device without electricity for 3 h and electric field treatment group (n=52) treated with 200 mV/mm electric field for 3 h (the same treatment methods below). The cell movement direction was observed in the living cell workstation and the movement velocity, trajectory velocity, and direction of cosθ of cell movement within 3 h of treatment were calculated. HaCaT cells were divided into simulated electric field group and electric field treatment 1 h group, electric field treatment 2 h group, and electric field treatment 3 h group which were treated with 200 mV/mm electric field for corresponding time. HaCaT cells were divided into simulated electric field group and 100 mV/mm electric field group, 200 mV/mm electric field group, and 300 mV/mm electric field group treated with electric field of corresponding intensities for 3 h. The protein expression of acetylated α-tubulin was detected by Western blotting (n=3). HaCaT cells were divided into simulated electric field group and electric field treatment group, and the protein expression of acetylated α-tubulin was detected and located by immunofluorescence method (n=3). Data were statistically analyzed with Kruskal-Wallis H test,Mann-Whitney U test, Bonferroni correction, one-way analysis of variance, least significant difference test, and independent sample t test. Results Within 3 h of treatment, compared with that in simulated electric field group, the cells in electric field treatment group had obvious tendency to move directionally, the movement velocity and trajectory velocity were increased significantly (with Z values of -8.53 and -2.05, respectively, P<0.05 or P<0.01), and the directionality was significantly enhanced (Z=-8.65, P<0.01). Compared with (0.80±0.14) in simulated electric field group, the protein expressions of acetylated α-tubulin in electric field treatment 1 h group (1.50±0.08) and electric field treatment 2 h group (1.89±0.06) were not changed obviously (P>0.05), while the protein expression of acetylated α-tubulin of cells in electric field treatment 3 h group (3.37±0.36) was increased significantly (Z=-3.06, P<0.05). After treatment for 3 h, the protein expressions of acetylated α-tubulin of cells in 100 mV/mm electric field group, 200 mV/mm electric field group, and 300 mV/mm electric field group were 1.63±0.05, 2.24±0.08, and 2.00±0.13, respectively, which were significantly more than 0.95±0.27 in simulated electric field group (P<0.01). Compared with that in 100 mV/mm electric field group, the protein expressions of acetylated α-tubulin in 200 mV/mm electric field group and 300 mV/mm electric field group were increased significantly (P<0.01); the protein expression of acetylated α-tubulin of cells in 300 mV/mm electric field group was significantly lower than that in 200 mV/mm electric field group (P<0.05). After treatment for 3 h, compared with that in simulated electric field group, the acetylated α-tubulin of cells had enhanced directional distribution and higher protein expression (t=5.78, P<0.01). Conclusions Bio-intensity electric field can induce the directional migration of HaCaT cells and obviously up-regulate the level of α-ubulin acetylation after treatment at 200 mV/mm bio-intensity electric field for 3 h.
- Skin,
- Cell movement,
- Tubulin,
- Acetylation,
- Bio-intensity electric field,
- Epidermal cells

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References

[1]	GradaA, Otero-VinasM, Prieto-CastrilloF, et al. Research techniques made simple: analysis of collective cell migration using the wound healing assay[J]. J Invest Dermatol, 2017, 137(2): e11-e16. DOI: 10.1016/j.jid.2016.11.020.
[2]	冀然,张泽,王文平,等.生物强度电场对人表皮细胞株 HaCaT和小鼠表皮细胞运动性及CD9表达的影响[J].中华烧伤杂志,2021,37(1):34-41.DOI: 10.3760/cma.j.cn501120-20200115-00023.
[3]	TaiG, TaiM, ZhaoM. Electrically stimulated cell migration and its contribution to wound healing[J/OL]. Burns Trauma, 2018, 6(1): 20[2022-10-23]. https://pubmed.ncbi.nlm.nih.gov/30003115/. DOI: 10.1186/s41038-018-0123-2.
[4]	王文平,冀然,张泽,等.生物强度电场对人皮肤成纤维细胞转化的调节作用[J].中华烧伤与创面修复杂志,2022,38(4):354-362.DOI: 10.3760/cma.j.cn501120-20210112-00017.
[5]	JiR,TengM,ZhangZ,et al.Electric field down-regulates CD9 to promote keratinocytes migration through AMPK pathway[J].Int J Med Sci,2020,17(7):865-873.DOI: 10.7150/ijms.42840.
[6]	LinBJ,TsaoSH,ChenA,et al.Lipid rafts sense and direct electric field-induced migration[J].Proc Natl Acad Sci U S A,2017,114(32):8568-8573.DOI: 10.1073/pnas.1702526114.
[7]	GarcinC,StraubeA.Microtubules in cell migration[J].Essays Biochem,2019,63(5):509-520.DOI: 10.1042/EBC20190016.
[8]	JankeC, MontagnacG. Causes and consequences of microtubule acetylation[J]. Curr Biol, 2017, 27(23):R1287-R1292. DOI: 10.1016/j.cub.2017.10.044.
[9]	MorelliG, EvenA, Gladwyn-NgI, et al. p27Kip1 modulates axonal transport by regulating α-tubulin acetyltransferase 1 stability[J]. Cell Rep, 2018, 23(8): 2429-2442. DOI: 10.1016/j.celrep.2018.04.083.
[10]	ChawanV, YevateS, GajbhiyeR, et al. Acetylation/deacetylation and microtubule associated proteins influence flagellar axonemal stability and sperm motility[J]. Biosci Rep, 2020, 40(12): BSR20202442. DOI: 10.1042/BSR20202442.
[11]	DeakinNO, TurnerCE. Paxillin inhibits HDAC6 to regulate microtubule acetylation, Golgi structure, and polarized migration[J]. J Cell Biol, 2014, 206(3): 395-413. DOI: 10.1083/jcb.201403039.
[12]	NuccitelliR. A role for endogenous electric fields in wound healing[J]. Curr Top Dev Biol, 2003, 58: 1-26. DOI: 10.1016/S0070-2153(03)58001-2.
[13]	CaiJ, ZhongY, TianS. Naturally occurring davanone terpenoid exhibits anticancer potential against ovarian cancer cells by inducing programmed cell death, by inducing caspase-dependent apoptosis, loss of mitochondrial membrane potential, inhibition of cell migration and invasion and targeting PI3K/AKT/MAPK signaling pathway[J]. J BUON, 2020, 25(5): 2301-2307.
[14]	GoodsonHV, JonassonEM. Microtubules and microtubule-associated proteins[J]. Cold Spring Harb Perspect Biol, 2018, 10(6): a022608. DOI: 10.1101/cshperspect.a022608.
[15]	LaFlammeSE, Mathew-SteinerS, SinghN, et al. Integrin and microtubule crosstalk in the regulation of cellular processes[J]. Cell Mol Life Sci, 2018, 75(22): 4177-4185. DOI: 10.1007/s00018-018-2913-x.
[16]	JankeC, MagieraMM. The tubulin code and its role in controlling microtubule properties and functions[J]. Nat Rev Mol Cell Biol, 2020, 21(6): 307-326. DOI: 10.1038/s41580-020-0214-3.
[17]	Roll-MecakA. The tubulin code in microtubule dynamics and information encoding[J]. Dev Cell, 2020, 54(1): 7-20. DOI: 10.1016/j.devcel.2020.06.008.
[18]	LiuN, XiongY, RenY, et al. Proteomic profiling and functional characterization of multiple post-translational modifications of tubulin[J]. J Proteome Res, 2015, 14(8): 3292-3304. DOI: 10.1021/acs.jproteome.5b00308.
[19]	XuZ, SchaedelL, PortranD, et al. Microtubules acquire resistance from mechanical breakage through intralumenal acetylation[J]. Science, 2017, 356(6335): 328-332. DOI: 10.1126/science.aai8764.
[20]	Eshun-WilsonL, ZhangR, PortranD, et al. Effects of α-tubulin acetylation on microtubule structure and stability[J]. Proc Natl Acad Sci U S A, 2019, 116(21): 10366-10371. DOI: 10.1073/pnas.1900441116.
[21]	BanceB, SeetharamanS, LeducC, et al. Microtubule acetylation but not detyrosination promotes focal adhesion dynamics and astrocyte migration[J]. J Cell Sci, 2019, 132(7): jcs225805. DOI: 10.1242/jcs.225805.
[22]	AtkinsonSJ, GontarczykAM, AlghamdiAA, et al. The β3-integrin endothelial adhesome regulates microtubule-dependent cell migration[J]. EMBO Rep, 2018, 19(7): e44578. DOI: 10.15252/embr.201744578.
[23]	MyatMM, RashmiRN, MannaD, et al. Drosophila KASH-domain protein Klarsicht regulates microtubule stability and integrin receptor localization during collective cell migration[J]. Dev Biol, 2015, 407(1): 103-114. DOI: 10.1016/j.ydbio.2015.08.003.
[24]	Rampioni VinciguerraGL, CitronF, SegattoI, et al. p27kip1 at the crossroad between actin and microtubule dynamics[J]. Cell Div, 2019, 14(1): 2. DOI: 10.1186/s13008-019-0045-9.
[25]	ReedNA, CaiD, BlasiusTL, et al. Microtubule acetylation promotes kinesin-1 binding and transport[J]. Curr Biol, 2006, 16(21): 2166-2172. DOI: 10.1016/j.cub.2006.09.014.
[26]	Castro-CastroA, JankeC, MontagnacG, et al. ATAT1/MEC-17 acetyltransferase and HDAC6 deacetylase control a balance of acetylation of alpha-tubulin and cortactin and regulate MT1-MMP trafficking and breast tumor cell invasion[J]. Eur J Cell Biol, 2012, 91(11/12): 950-960. DOI: 10.1016/j.ejcb.2012.07.001.
[27]	van DijkJ, BompardG, CauJ, et al. Microtubule polyglutamylation and acetylation drive microtubule dynamics critical for platelet formation[J]. BMC Biol, 2018, 16(1): 116. DOI: 10.1186/s12915-018-0584-6.
[28]	ShiP, WangY, HuangY, et al. Arp2/3-branched actin regulates microtubule acetylation levels and affects mitochondrial distribution[J]. J Cell Sci, 2019, 132(6): jcs226506. DOI: 10.1242/jcs.226506.
[29]	HubbertC, GuardiolaA, ShaoR, et al. HDAC6 is a microtubule-associated deacetylase[J]. Nature, 2002, 417(6887): 455-458. DOI: 10.1038/417455a.
[30]	AdalbertR, KaiedaA, AntoniouC, et al. Novel HDAC6 inhibitors increase tubulin acetylation and rescue axonal transport of mitochondria in a model of charcot-marie-tooth type 2F[J]. ACS Chem Neurosci, 2020, 11(3): 258-267. DOI: 10.1021/acschemneuro.9b00338.
[31]	KershawS, MorganDJ, BoydJ, et al. Glucocorticoids rapidly inhibit cell migration through a novel, non-transcriptional HDAC6 pathway[J]. J Cell Sci, 2020, 133(11): jcs242842. DOI: 10.1242/jcs.242842.
[32]	Valenzuela-FernándezA, CabreroJR, SerradorJM, et al. HDAC6: a key regulator of cytoskeleton, cell migration and cell-cell interactions[J]. Trends Cell Biol, 2008, 18(6): 291-297. DOI: 10.1016/j.tcb.2008.04.003.
[33]	KeB, ChenY, TuW, et al. Inhibition of HDAC6 activity in kidney diseases: a new perspective[J]. Mol Med, 2018, 24(1): 33. DOI: 10.1186/s10020-018-0027-4.
[34]	KalinskiAL, KarAN, CraverJ, et al. Deacetylation of Miro1 by HDAC6 blocks mitochondrial transport and mediates axon growth inhibition[J]. J Cell Biol, 2019, 218(6): 1871-1890. DOI: 10.1083/jcb.201702187.
[35]	SeidelC, SchnekenburgerM, DicatoM, et al. Histone deacetylase 6 in health and disease[J]. Epigenomics, 2015, 7(1): 103-118. DOI: 10.2217/epi.14.69.
[36]	Shafaq-ZadahM, Gomes-SantosCS, BardinS, et al. Persistent cell migration and adhesion rely on retrograde transport of β1 integrin[J]. Nature Cell Biology, 2016, 18(1): 54-64. DOI: 10.1038/ncb3287.
[37]	RenX, SunH, LiuJ, et al. Keratinocyte electrotaxis induced by physiological pulsed direct current electric fields[J]. Bioelectrochemistry, 2019, 127: 113-124. DOI: 10.1016/j.bioelechem.2019.02.001.