Experimental Study of lncRNA MALAT1 on Proliferation and Osteogenic Differentiation of Human Periodontal Ligament Stem Cells by Regulating miR-150-5p/XBP1 Axis
10.3969/j.issn.1671-7414.2025.04.014
- VernacularTitle: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
- Author:
Qian ZHANG
1
;
Xinyi TANG
1
;
Lie WANG
1
Author Information
1. 武汉市第三医院口腔科,武汉 430000
- Publication Type:Journal Article
- Keywords:
long non coding RNA;
metastasis associated lung adenocarcinoma transcript 1;
microRNA-150-5p;
periodontal ligament stem cells;
X-box binding protein 1;
osteogenic differentiation
- From:
Journal of Modern Laboratory Medicine
2025;40(4):79-85
- CountryChina
- Language:Chinese
-
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.
