hsa-let-7a-5p调控牙周膜干细胞增殖及凋亡

张晔琳; 马丽娅; 彭旭晖; 杨禾丰; 佘睿

doi:10.12259/j.issn.2095-610X.S20231028

hsa-let-7a-5p调控牙周膜干细胞增殖及凋亡

doi: 10.12259/j.issn.2095-610X.S20231028

张晔琳^{1, 2},
马丽娅^{1, 2},
彭旭晖³,
杨禾丰^{1, 2},
佘睿^{1, 2, 4, ,}

1.
昆明医科大学口腔医学院，云南昆明　650500
2.
云南省口腔医学重点实验室，云南昆明　650500
3.
永善县人民医院，云南昭通　657300
4.
昆明医科大学口腔医院牙体牙髓科，云南昆明　650500

基金项目: 国家级大学生创新创业训练计划基金资助项目（202110678007）；云南省教育厅科学研究基金指导性项目（42017002）；云南省教育厅科学研究基金资助项目（2022Y216）

详细信息

作者简介:
张晔琳（2001～），女，云南保山人，在读本科生

通讯作者:
佘睿，E-mail：3116964506@QQ.com

中图分类号: R781.4
计量
- 文章访问数: 1463
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- PDF下载量: 17
- 被引次数: 0
出版历程
- 收稿日期: 2023-08-13
- 网络出版日期: 2023-09-12
- 刊出日期: 2023-10-25

Hsa-let-7a-5p Regulates Proliferation and Apoptosis of Periodontal Ligament Stem Cells

Yelin ZHANG^{1, 2},
Liya MA^{1, 2},
Xuhui PENG³,
Hefeng YANG^{1, 2},
Rui SHE^{1, 2, 4
, ,}

1.
School of Stomatology，Kunming Medical University，Kunming Yunnan 650500
2.
Yunnan Key Laboratory of Stomatology，Kunming Yunnan 650500
3.
Yongshan County People’s Hospital，Zhaotong Yunnan 657300
4.
Dept. of Endodontology，The Affiliated Stomatology Hospital of Kunming Medical University，Kunming Yunnan 650500，China

摘要

摘要: 目的探讨hsa-let-7a-5p对牙周膜干细胞（periodontal ligament stem cells，PDLSCs）增殖及凋亡的影响。方法将let-7a-5p mimics、mimics nc、let-7a-5p inhibitor、inhibitor nc转染进入PDLSCs，通过RT-qPCR测定转染效率，通过CCK-8、细胞周期、EdU实验检测let-7a-5p对PDLSCs增殖的影响，通过细胞凋亡实验检测PDLSCs的凋亡率。结果 let-7a-5p成功转染进入PDLSCs。let-7a-5p mimics可抑制PDLSCs增殖能力（P < 0.05），促进PDLSCs凋亡（P < 0.01）。let-7a-5p inhibitor可促进PDLSCs增殖能力（P < 0.05），抑制PDLSCs凋亡（P < 0.01）。结论 hsa-let-7a-5p可抑制PDLSCs增殖，促进PDLSCs凋亡。
- hsa-let-7a-5p /
- microRNA /
- 牙周膜干细胞 /
- 增殖 /
- 凋亡
Abstract: Objective To investigate the effect of hsa-let-7a-5p on the proliferation and apoptosis of periodontal ligament stem cells （PDLSCs）. Methods let-7a-5p mimics, mimics nc, let-7a-5p inhibitor, and inhibitor nc were transfected into PDLSCs, and transfection efficiency was measured by RT-qPCR. The effects of let-7a-5p on PDLSCs proliferation were evaluated by CCK-8, cell cycle, and EdU experiments, while the apoptosis rate of PDLSCs was detected by apoptosis experiments. Results let-7a-5p was successfully transfected into PDLSCs. let-7a-5p mimics inhibited cell proliferation （P < 0.05） and promoted cell apoptosis of PDLSCs （P < 0.01）. let-7a-5p inhibitor promoted cell proliferation （P < 0.05） and inhibited cell apoptosis of PDLSCs （P < 0.01）. Conclusion hsa-let-7a-5p can inhibit the proliferation of PDLSCs and promote the apoptosis of PDLSCs.
- hsa-let-7a-5p /
- microRNA /
- Periodontal ligament stem cell /
- Proliferation /
- Apoptosis

HTML全文

图 1 let-7a-5p转染效率测定

A：荧光验证let-7a-5p转染进入PDLSCs；B：RT-qPCR测定let-7a-5p转染效率。n = 3，^**P < 0.01，^#P < 0.0001。

Figure 1. Transfection efficiency of let-7a-5p

下载: 全尺寸图片幻灯片

图 2 let-7a-5p抑制牙周膜干细胞增殖

A：CCK-8探究let-7a-5p mimics对PDLSCs增殖的影响，n = 4；B：CCK-8探究let-7a-5p inhibitor对PDLSCs增殖的影响，n = 4；C：EdU实验探究let-7a-5p mimics对PDLSCs增殖的影响，n = 5；D：EdU实验探究let-7a-5p inhibitor对PDLSCs增殖的影响，n = 5；E：细胞周期探究let-7a-5p mimics对PDLSCs增殖的影响；F：细胞周期探究let-7a-5p inhibitor对PDLSCs增殖的影响。^*P < 0.05，^**P < 0.01，^***P < 0.001，^#P < 0.0001。

Figure 2. let-7a-5p inhibits periodontal ligament stem cell proliferation

下载: 全尺寸图片幻灯片

图 3 let-7a-5p促进牙周膜干细胞凋亡

A：细胞凋亡实验探究let-7a-5p mimics对PDLSCs凋亡的影响；B：细胞凋亡实验探究let-7a-5p inhibitor对PDLSCs凋亡的影响。n = 3，^**P < 0.01。

Figure 3. let-7a-5p promotes periodontal ligament stem cell apoptosis

下载: 全尺寸图片幻灯片

图 4 let-7a-5p靶基因及功能预测

A：let-7a-5p靶基因预测的韦恩图；B：靶基因的GO富集气泡图；C：靶基因KEGG通路富集气泡图。

Figure 4. let-7a-5p target gene and function prediction

下载: 全尺寸图片幻灯片

参考文献(31)

[1]	Kwon T,Lamster I B,Levin L. Current concepts in the management of periodontitis[J]. International Dental Journal,2021,71(6):462-476. doi: 10.1111/idj.12630
[2]	Isola G,Santonocito S,Lupi S M,et al. Periodontal health and disease in the context of systemic diseases[J]. Mediators of Inflammation,2023,2023:9720947.
[3]	Păunică I,Giurgiu M,Dumitriu AS,et al. The bidirectional relationship between periodontal disease and diabetes mellitus-A review[J]. Diagnostics (Basel),2023,13(4):681. doi: 10.3390/diagnostics13040681
[4]	Seo B M,Miura M,Gronthos S,et al. Investigation of multipotent postnatal stem cells from human periodontal ligament[J]. Lancet,2004,364(9429):149-155. doi: 10.1016/S0140-6736(04)16627-0
[5]	Zhao X,Lin H,Ding T,et al. Overview of the main biological mechanisms linked to changes in periodontal ligament stem cells and the inflammatory microenvironment[J]. J Zhejiang Univ Sci B,2023,24(5):373-386. doi: 10.1631/jzus.B2200576
[6]	Luan X,Zhou X,Trombetta-eSilva J,et al. MicroRNAs and periodontal homeostasis[J]. J Dent Res,2017,96(5):491-500. doi: 10.1177/0022034516685711
[7]	Huang P,Jia L. MicroRNA-28-5p as a potential diagnostic biomarker for chronic periodontitis and its role in cell proliferation and inflammatory response[J]. J Dent Sci,2022,17(4):1501-1509. doi: 10.1016/j.jds.2022.04.031
[8]	Venugopal P,Koshy T,Lavu V,et al. Differential expression of microRNAs let-7a,miR-125b,miR-100,and miR-21 and interaction with NF-kB pathway genes in periodontitis pathogenesis[J]. J Cell Physiol,2018,233(8):5877-5884. doi: 10.1002/jcp.26391
[9]	Lee Y H,Na H S,Jeong S Y,et al. Comparison of inflammatory microRNA expression in healthy and periodontitis tissues[J]. Biocell,2011,35(2):43-49. doi: 10.32604/biocell.2011.35.043
[10]	Lee N H,Lee E,Kim Y S,et al. Differential expression of microRNAs in the saliva of patients with aggressive periodontitis: A pilot study of potential biomarkers for aggressive periodontitis[J]. J Periodontal Implant Sci,2020,50(5):281-290. doi: 10.5051/jpis.2000120006
[11]	Ma L,Rao N,Jiang H,et al. Small extracellular vesicles from dental follicle stem cells provide biochemical cues for periodontal tissue regeneration[J]. Stem Cell Res Ther,2022,13(1):92. doi: 10.1186/s13287-022-02767-6
[12]	Li Q,Yang G,Li J,et al. Stem cell therapies for periodontal tissue regeneration: A network meta-analysis of preclinical studies[J]. Stem Cell Res Ther,2020,11(1):427. doi: 10.1186/s13287-020-01938-7
[13]	Tomokiyo A,Wada N,Maeda H. Periodontal ligament stem cells: Regenerative potency in periodontium[J]. Stem Cells Dev,2019,28(15):974-985. doi: 10.1089/scd.2019.0031
[14]	Jin H Y,Gonzalez-Martin A,Miletic A V,et al. Transfection of microRNA mimics should be used with caution[J]. Front Genet,2015,6:340.
[15]	Esau C C. Inhibition of microRNA with antisense oligonucleotides[J]. Methods,2008,44(1):55-60. doi: 10.1016/j.ymeth.2007.11.001
[16]	Robertson B,Dalby A B,Karpilow J,et al. Specificity and functionality of microRNA inhibitors[J]. Silence,2010,1(1):10. doi: 10.1186/1758-907X-1-10
[17]	Campani V,De Rosa G,Misso G,et al. Lipid nanoparticles to deliver miRNA in cancer[J]. Curr Pharm Biotechnol,2016,17(8):741-749. doi: 10.2174/138920101708160517234941
[18]	Johnson C D,Esquela-Kerscher A,Stefani G,et al. The let-7 microRNA represses cell proliferation pathways in human cells[J]. Cancer Res,2007,67(16):7713-7722. doi: 10.1158/0008-5472.CAN-07-1083
[19]	Chen Z,Qiu J,Gao Y,et al. Study on the mechanism of let-7a-5p in regulating the proliferation in cervical cancer cells[J]. Clin Transl Oncol,2022,24(8):1631-1642. doi: 10.1007/s12094-022-02810-1
[20]	Yu J J,Pi W S,Cao Y,et al. Let-7a inhibits osteosarcoma cell growth and lung metastasis by targeting Aurora-B[J]. Cancer Manag Res,2018,10:6305-6315. doi: 10.2147/CMAR.S185090
[21]	Liu C,Chen Z,Fang M,et al. MicroRNA let-7a inhibits proliferation of breast cancer cell by downregulating USP32 expression[J]. Transl Cancer Res,2019,8(5):1763-1771. doi: 10.21037/tcr.2019.08.30
[22]	Chen Y,Qiao L,Zhang Z,et al. Let-7a inhibits proliferation and promotes apoptosis of human asthmatic airway smooth muscle cells[J]. Exp Ther Med,2019,17(5):3327-3334.
[23]	James N,Kini S,Pai S,et al. Comparative evaluation of corneal storage medias used as tooth avulsion medias in maintaining the viability of periodontal ligament cells using the cell counting kit-8 assay[J]. Clin Cosmet Investig Dent,2022,14:87-94. doi: 10.2147/CCIDE.S314478
[24]	Sun X,Zhang C,Jin H,et al. Flow cytometric analysis of T lymphocyte proliferation in vivo by EdU incorporation[J]. Int Immunopharmacol,2016,41:56-65. doi: 10.1016/j.intimp.2016.10.019
[25]	Rieger A M. Flow cytometry and cell cycle analysis: An overview[J]. Methods Mol Biol,2022,2579:47-57.
[26]	Xu X,Lai Y,Hua Z C. Apoptosis and apoptotic body: Disease message and therapeutic target potentials[J]. Biosci Rep,2019,39(1):BSR20180992. doi: 10.1042/BSR20180992
[27]	Zhang J,Dong X,Yan Q,et al. Galectin-1 inhibited LPS-induced autophagy and apoptosis of human periodontal ligament stem cells[J]. Inflammation,2021,44(4):1302-1314. doi: 10.1007/s10753-021-01417-y
[28]	Liu L,Zhao J,Peng Y,et al. miR-let-7a-5p inhibits invasion and migration of hepatoma cells by regulating BZW2 expression[J]. Onco Targets Ther,2020,13:12269-12279. doi: 10.2147/OTT.S278954
[29]	Cai K,Wan Y,Sun G,et al. Let-7a inhibits proliferation and induces apoptosis by targeting EZH2 in nasopharyngeal carcinoma cells[J]. Oncol Rep,2012,28(6):2101-2106. doi: 10.3892/or.2012.2027
[30]	Ersahin T,Tuncbag N,Cetin-Atalay R. The PI3K/AKT/mTOR interactive pathway[J]. Mol Biosyst,2015,11(7):1946-1954. doi: 10.1039/C5MB00101C
[31]	Xia Z,Li Q,Tang Z. Network pharmacology,molecular docking,and experimental pharmacology explored Ermiao wan protected against periodontitis via the PI3K/AKT and NF-κB/MAPK signal pathways[J]. J Ethnopharmacol,2023,303:115900. doi: 10.1016/j.jep.2022.115900

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