[1]张 千,唐欣轶,汪 烈.lncRNA MALAT1调节miR-150-5p/XBP1轴对人牙周膜干细胞增殖和成骨分化的实验研究[J].现代检验医学杂志,2025,40(04):79-85.[doi:10.3969/j.issn.1671-7414.2025.04.014]
　ZHANG Qian,TANG Xinyi,WANG Lie.Experimental Study of lncRNA MALAT1 on Proliferation and Osteogenic Differentiation of Human Periodontal Ligament Stem Cells by Regulating miR-150-5p/XBP1 Axis[J].Journal of Modern Laboratory Medicine,2025,40(04):79-85.[doi:10.3969/j.issn.1671-7414.2025.04.014]
点击复制

lncRNA MALAT1调节miR-150-5p/XBP1轴对人牙周膜干细胞增殖和成骨分化的实验研究()

《现代检验医学杂志》[ISSN:/CN:]

卷:
第40卷
期数:
2025年04期
页码:
79-85
栏目:
论著
出版日期:
2025-07-15

文章信息/Info

Title:
Experimental Study of lncRNA MALAT1 on Proliferation and Osteogenic Differentiation of Human Periodontal Ligament Stem Cells by Regulating miR-150-5p/XBP1 Axis
文章编号:
1671-7414（2025）04-079-07
作者:
张 千，唐欣轶，汪 烈
（武汉市第三医院口腔科，武汉 430000）
Author(s):
ZHANG Qian, TANG Xinyi, WANG Lie
(Department of Stomatology, Wuhan Third Hospital, Wuhan 430000，China)
关键词:
长链非编码RNA; 肺腺癌转移相关转录子1; 微小RNA-150-5p; 牙周膜干细胞; X-框结合蛋白1; 成骨分化
分类号:
R781.42；R392.12
DOI:
10.3969/j.issn.1671-7414.2025.04.014
文献标志码:
A
摘要:
目的 探讨长链非编码RNA（lncRNA）肺腺癌转移相关转录子1（MALAT1）调节miR-150-5p/X- 框结合蛋白1（XBP1）轴对人牙周膜干细胞（PDLSCs）增殖和成骨分化的影响及机制。方法 实时荧光定量PCR（qRT-PCR）检测未分化及已分化PDLSCs 中lncRNA MALAT1，miR-150-5p 及XBP1 mRNA 表达。取传至第三代的PDLSCs，将其分为对照组、pcDNA 组、pcDNA-lncRNA MALAT1 组、anti-miR-NC 组、anti-miR-150-5p 组、pcDNA-lncRNAMALAT1+mimic NC 组和pcDNA-lncRNA MALAT1+miR-150-5p mimic 组。qRT-PCR 检测lncRNA MALAT1，miR-150-5p 及XBP1 mRNA 在PDLSCs 中的表达；CCK-8，克隆形成实验检测PDLSCs 增殖；对硝基酚磷酸底物法检测碱性磷酸酶（ALP）活性；茜素红染色检测矿化结节形成率；Western blot 检测XBP1，细胞周期素D1（Cyclin D1）、骨钙素（OCN）、Runt 相关转录因子2（RUNX2）和骨桥蛋白（OPN）表达；双荧光素酶实验验证lncRNA MALAT1 与miR-150-5p 及miR-150-5p 与XBP1的靶向关系。 结果 与未分化组比较，已分化组PDLSCs中lncRNA MALAT1表达（1.95±0.14vs 1.00±0.00）及XBP1 mRNA 表达（1.63±0.12 vs 1.00±0.00）升高，miR-150-5p 表达（0.26±0.01 vs 1.00±0.00）降低，差异具有统计学意义（t=16.622，12.860，181.262，均P<0.001）。过表达lncRNA MALAT1 或下调miR-150-5p 均可促进人PDLSCs 增殖和成骨分化（t=13.693 ～ 45.518），miR-150-5p mimic 逆转了lncRNA MALAT1 过表达对PDLSCs 增殖及成骨分化的促进作用（t= 9.229 ～ 27.854），差异具有统计学意义（均P<0.05）。双荧光素酶实验证实，miR-150-5P 过表达降低了转染lncRNA MALATL-WT，XBP1-WT 细胞的荧光素酶活性（t=56.546，89.826，均P ＜ 0.001）。结论 过表达lncRNA MALAT1 可能通过下调miR-150-5p 促进XBP1 表达进而促进人PDLSCs 增殖和成骨分化。
Abstract:
Objective To investigate the effects of long non coding RNA (lncRNA) metastasis associated lung adenocarcinoma transcript 1 (MALAT1) on the proliferation and osteogenic differentiation of human periodontal ligament stem cells (PDLSCs) by regulating the miR-150-5p/X-box binding protein 1 (XBP1) axis. Methods Quantitative real-time PCR (qRT-PCR) was used to detect the expression of lncRNA MALAT1, miR-150-5p and XBP1 mRNA in undifferentiated and differentiated PDLSCs. The third generation PDLSCs were divided into control group, pcDNA group, pcDNA-lncRNA MALAT1 group, anti-miR-NC group, anti-miR-150-5p group, pcDNA-lncRNA MALAT1+mimic NC group and pcDNA-lncRNA MALAT1+miR-150-5p mimic group. The expression of lncRNA MALAT1, miR-150-5p and XBP1 mRNA of PDLSCs were detected by qRT-PCR. CCK-8 and the clone formation assay were used to detect the proliferation of PDLSCs. P-nitrophenolphosphate substrate method was used to detect alkaline phosphatase (ALP) activity. Alizarin red staining was applied to detect the formation rate of mineralized nodules. Western blot was used to detect XBP1, Cyclin D1, osteocalcin (OCN), Runt related transcription factor 2 (RUNX2) and osteopontin (OPN) proteins. Dual luciferase assay was used to verify the targeting relationship between lncRNA MALAT1 and miR-150-5p, and between miR-150-5p and XBP1 was validated. Results Compared with the undifferentiated group, the expression of lncRNA MALAT1 (1.95±0.14 vs 1.00±0.00) and XBP1 mRNA expression (1.63±0.12 vs 1.00±0.00) increased in the differentiated PDLSCs, the expression of miR-150-5p decreased (0.26±0.01 vs 1.00±0.00) in differentiated group, with the differences were statistical significance (t=16.622, 12.860, 181.262，all P<0.001). Overexpression of lncRNA MALAT1 or down-regulation of miR-150-5p could promote the proliferation and osteogenic differentiation of human PDLSCs (t= 13.693 ～ 45.518)，miR-150- 5p mimic reversed the effects of overexpression of lncRNA MALAT1 on proliferation and osteogenic differentiation of PDLSCs (t=9.229 ～ 27.854)，and the differences were statistically significant (all P<0.05)，respectively. Dual luciferase assay confirmed that overexpression of miR-150-5P reduced the luciferase activity in transfected lncRNA MALAT1-WT and XBP1-WT cells (t=56.546, 89.826, all P<0.001). Conclusion Overexpression of lncRNA MALAT1 may promote XBP1 expression by downregulating miR-150-5p, thereby promoting proliferation and osteogenic differentiation of human PDLSCs.

参考文献/References:

[1] ZHAO Ying, QUAN Yue, LEI Ting, et al. The role of inflammasome NLPR3 in the development and therapy of periodontitis[J]. International Journal of Medical Sciences, 2022, 19(10): 1603-1614.
[2] SUN Xuefei, LI Zhidan, DONG Shaojie, et al. Long non-coding RNA SNHG5 promotes osteogenic differentiation of human periodontal ligament stem cells via mediating miR-23b-3p/Runx2 axis[J]. International Journal of Medical Sciences, 2023, 20(7): 958-968.
[3] HAN Ruiqi, ZHANG Wenjuan, ZHANG Lina, et al. Notoginsenoside R1 promotes proliferation and osteogenic differentiation of hPDLSCs via Wnt/β-catenin signaling pathway [J].Drug Design,Development and Therapy, 2022, 16: 4399-4409.
[4] WANG Hong, GAO Shuting, DISSANAYAKA W L. Circ_0003764 regulates the osteogenic differentiation of periodontal ligament stem cells[J].International Dental Journal, 2024, 74(5): 1110-1119.
[5] ZHOU Yuan, XU Zhuo, WANG Yuanyi, et al. LncRNA MALAT1 mediates osteogenic differentiation in osteoporosis by regulating the miR-485-5p/WNT7B axis[J]. Frontiers in Endocrinology (Lausanne), 2023,13: 922560.
[6] GU Nan,WANG Yao,LI Lingfeng, et al.The mechanism of lncRNA MALAT1 targeting the miR-124-3p/IGF2BP1 axis to regulate osteogenic differentiation of periodontal ligament stem cells[J]. Clinical Oral Investigations, 2024, 28(4): 219.
[7] 陈静涛, 王燕, 李宝东, 等.MiR-150-5p 下调通过激活Wnt 信号通路促进人牙周膜干细胞成骨分化及MMP2 表达[J]. 临床与病理杂志,2021,41(12):2763-2770. CHEN Jingtao, WANG Yan, LI Baodong, et al. MiR-150-5p downregulation facilitates osteogenic differentiation of human periodontal ligament stem cells and MMP2 expression via activating Wnt signaling pathway[J]. Journal of Clinical and Pathological Research, 2021, 41(12): 2763-2770.
[8] CUI Ziwei, QIN Ruoshan, FENG Jianbao, et al. XBP1s gene of endoplasmic reticulum stress enhances proliferation and osteogenesis of human periodontal ligament cells [J]. Tissue and Cell, 2023, 83: 102139.
[9] TELES F, COLLMAN R G, MOMINKHAN D,et al. Viruses, periodontitis, and comorbidities[J]. Periodontology 2000, 2022, 89(1): 190-206.
[10] HERRERA D,SANZ M,KEBSCHULL M, et al. Treatment of stage Ⅳ periodontitis: the EFP S3 level clinical practice guideline[J]. Journal of Clinical Periodontology, 2022, 49 (Suppl 22): 4-71.
[11] MOLINA A, SANZ-S?NCHEZ I, SANZ-MART?N I, et al. Complications in sinus lifting procedures:classification and management[J]. Periodontology 2000, 2022, 88(1): 103-115.
[12] MOHEBICHAMKHORAMI F, FATTAHI R, NIKNAM Z, et al. Periodontal ligament stem cells as a promising therapeutic target for neural damage[J]. Stem Cell Research & Therapy, 2022, 13(1): 273.
[13] ZHANG Yunfan, DOU Xueyu, ZHANG Lingyun, et al. Facile fabrication of a biocompatible composite gel with sustained release of aspirin for bone regeneration [J]. Bioactive Materials, 2021, 11: 130-139.
[14] TU Shaoqin, CHEN Yihua, FENG Yi, et al. LncRNA CYTOR facilitates osteogenic differentiation of human periodontal ligament stem cells by modulating SOX11 via sponging miR-6512-3p [J]. Stem Cells International,2023, 2023: 5671809.
[15] HUA Li, ZHANG Xiaohong. MALAT1 regulates osteogenic differentiation of human periodontal ligament stem cells through mediating miR-155-5p/ETS1 axis [J]. Tissue and Cell, 2021, 73: 101619.
[16] CHEN Pei, HUANG Yanhong, WANG Yarong, et al. MALAT1 overexpression promotes the proliferation of human periodontal ligament stem cells by upregulating fibroblast growth factor 2[J]. Experimental and Therapeutic Medicine, 2019, 18(3): 1627-1632.
[17] GU Yingzhi, BAI Yuxing. LncRNA MALAT1 promotes osteogenic differentiation through the miR-93-5p/SMAD5 axis[J]. Oral Diseases, 2024, 30(4): 2398-2409.
[18] YING Ming, ZHANG Bin. Daidzein promotes the proliferation and osteogenic differentiation of periodontal ligament stem cell[J]. Oral Diseases, 2023,29(3): 1226-1233.
[19] FAN Chun, LI Zhiyuan, JI Qiuxia, et al. Carboxymethyl chitin or chitosan for osteoinduction effect on the human periodontal ligament stem cells[J]. Dental Materials Journal, 2022, 41(3): 392-401.
[20] ZHANG Ying, WANG Fuyou, CHEN Guangxing,et al. LncRNA MALAT1 promotes osteoarthritis by modulating miR-150-5p/AKT3 axis [J]. Cell & Bioscience, 2019, 9: 54.
[21] HUANG Jing, XU Yuejing, HUANG Pengcheng. Salivary miR-150-5p as an indicator of periodontitis severity and regulator of human periodontal ligament fibroblast behavior by targeting AIFM2[J]. Journal of Periodontal Research, 2024, 59(1): 187-194.
[22] 廖烘, 刘伟, 龙运峰.LncRNA NEAT1 通过调节miR-125b-5p/IGFBP5 轴对血管瘤内皮细胞增殖、凋亡、迁移的实验研究[J]. 现代检验医学杂志,2023,38(3):65-71. LIAO Hong, LIU Wei, LONG Yunfeng. Experimental study on lncRNA NEAT1 on proliferation, apoptosis and migration of hemangioma endothelial cells by regulating miR-125b-5p/IGFBP5 axis[J]. Journal of Modern Laboratory Medicine, 2023, 38(3): 65-71.
[23] 陆佳艺, 伍倩琪, 陈伊燕, 等. 牙龈卟啉单胞菌来源的脂多糖通过X 盒结合蛋白1 调控脂肪细胞胰岛素信号通路的机制研究[J]. 华西口腔医学杂志,2022,40(2):148-154. LU Jiayi, WU Qianqi, CHEN Yiyan, et al. Mechanism of porphyromonas gingivalis-lipopolysaccharide in regulating the insulin signaling pathway in adipo-cytes via x-box binding protein 1[J]. West China Journal of Stomatology, 2022, 40(2): 148-154.

相似文献/References:

[1]张 振,李 芬.ceRNA与肿瘤[J].现代检验医学杂志,2016,31(04):128.[doi:10.3969/j.issn.1671-7414.2016.04.036]
[2]白盈盈,朱光旭,潘兴华.长链非编码RNA对结直肠癌潜在诊断价值的研究进展[J].现代检验医学杂志,2018,33(01):161.[doi:10.3969/j.issn.1671-7414.2018.01.001]
[3]洪　宏,袁建芬,喻海忠.乳腺癌患者血清长链非编码RNA ATB 表达水平检测及临床诊断价值[J].现代检验医学杂志,2020,35(02):22.[doi:10.3969 ／ j.issn.1671-7414.2020.02.007]
　HONG Hong,YUAN Jian-fen,YU Hai-zhong.Detection and Clinical Diagnosis Value of Serum Long Non-codingRNA ATB in Patients with Breast Cancer[J].Journal of Modern Laboratory Medicine,2020,35(04):22.[doi:10.3969 ／ j.issn.1671-7414.2020.02.007]
[4]李　悦,徐焕铭,樊　华.基于TCGA 数据对60 岁以上不同分层急性髓系白血病患者相关lncRNA 的基因信息学分析[J].现代检验医学杂志,2020,35(03):20.[doi:10.3969/j.issn.1671-7414.2020.03.005]
　LI Yue,XU Huan-ming,FAN Hua.Genetic Informatics Analysis of lncRNA Related to Patients with Different Stratified Acute Myeloid Leukemia Over 60 Years Based on TCGA Database[J].Journal of Modern Laboratory Medicine,2020,35(04):20.[doi:10.3969/j.issn.1671-7414.2020.03.005]
[5]梁晓虎,郭俊霞,雷尚昆.食管癌组织中lnc RNA DNAJC3-AS1的表达及其与临床病理特征的相关性研究[J].现代检验医学杂志,2021,36(02):75.[doi:doi:10.3969/j.issn.1671-7414.2021.02.018]
　LIANG Xiao-hu,GUO Jun-xia,LEI Shangkun.Expression of Lnc RNA DNAJC3-AS1 in Esophageal Carcinoma Tissues and Its Correlation with Clinicopathological Features[J].Journal of Modern Laboratory Medicine,2021,36(04):75.[doi:doi:10.3969/j.issn.1671-7414.2021.02.018]
[6]刘　霄,黄晓燕,王建华.长链非编码RNASNHG9在不同肿瘤中的最新研究进展[J].现代检验医学杂志,2021,36(04):176.[doi:10.3969/j.issn.1671-7414.2021.04.038]
　LIU Xiao,HUANG Xiao-yan,WANG Jian-hua.Reasearch Progress of Long Non-coding RNA SNHG9 in Different Tumors[J].Journal of Modern Laboratory Medicine,2021,36(04):176.[doi:10.3969/j.issn.1671-7414.2021.04.038]
[7]朱　娇,孙国才,刘素荣,等.食管癌患者血清神经元特异性烯醇化酶与长链非编码RNA ITGA9-AS1水平联合检测的实验诊断价值研究[J].现代检验医学杂志,2021,36(05):62.[doi:10.3969/j.issn.1671-7414.2021.05.014]
　ZHU Jiao,SUN Guo-cai,LIU Su-rong,et al.Experimental Diagnostic Value of Combined Detection of SerumNeuron-Specific Enolase and Long Non-Coding RNA ITGA9-AS1 inPatients with Esophageal Cancer[J].Journal of Modern Laboratory Medicine,2021,36(04):62.[doi:10.3969/j.issn.1671-7414.2021.05.014]
[8]刘立民,刘雪香,邹　燕,等.长链非编码RNA调控CD4+T淋巴细胞增殖分化和在免疫性疾病中的最新研究进展[J].现代检验医学杂志,2021,36(06):200.[doi:10.3969/j.issn.1671-7414.2021.06.044]
　LIU Li-min,LIU Xue-xiang,ZOU Yan,et al.Roles of lncRNA in Regulating the Proliferation, Differentiation of CD4+T Lymphocytes and Immune Diseases[J].Journal of Modern Laboratory Medicine,2021,36(04):200.[doi:10.3969/j.issn.1671-7414.2021.06.044]
[9]张文娟,郭 逸,段 勇,等.健康献血者机采血小板储存中LncRNA LIPCAR 的表达及初步研究[J].现代检验医学杂志,2023,38(03):119.[doi:10.3969/j.issn.1671-7414.2023.03.021]
　ZHANG Wen-juan,GUO Yi,DUAN Yong,et al.Expression and Preliminary Study of LncRNA LIPCAR in Platelet Storage of Mechanically Collected Platelets from Healthy Blood Donors[J].Journal of Modern Laboratory Medicine,2023,38(04):119.[doi:10.3969/j.issn.1671-7414.2023.03.021]
[10]李 轶,尚 立.非酒精性脂肪肝病患者血清LncRNA PVT1 表达水平与胰岛素抵抗及肝纤维化的相关性研究[J].现代检验医学杂志,2023,38(05):80.[doi:10.3969/j.issn.1671-7414.2023.05.015]
　LI Yi,SHANG Li.Correlation of Serum LncRNA PVT1 Expression Level with Insulin Resistance and Liver Fibrosis in Patients with Nonalcoholic Fatty Liver Disease[J].Journal of Modern Laboratory Medicine,2023,38(04):80.[doi:10.3969/j.issn.1671-7414.2023.05.015]

备注/Memo

备注/Memo:
基金项目：武汉市医学科研项目[ 健康发展（西医类）面上项目]（WX23A03）。
作者简介：张千（1993-），女，硕士，医师，研究方向：口腔医学，E-mail：qq13if7@163.com。
通讯作者：汪烈（1982-），男，硕士，主任医师，研究方向：口腔医学，E-mail：4325130@qq.com。
更新日期/Last Update: 2025-07-15