miR-431-5p靶向Smad4影响人牙髓干细胞增殖、分化的实验研究

Abstract

Abstract: Objective:To explore the regulation effect and mechanism of miR-431-5p on the proliferation and differentiation of human dental pulp stem cells (DPSC). Methods:The third molars of healthy people were collected to isolate and culture DPSC. In the experiments, mimic-NC group, miR-431-5p mimic group, pc-Smad4 group and pc-Smad4+miR-431-5p mimic group were set up. DPSC were transfected with lipid plastid transfection. The targeted relationship between miR-431-5p and Smad4 was verified by double luciferase reporter gene assay. The cells proliferation was detected by CCK-8. The calcified nodules were observed by alizarin red staining. The expression and activity of alkaline phosphatase (ALP) were detected by ALP staining kits and ALP activity test. The expressions of osteogenic differentiation marker osteocalcin (OCN) and Smad4 were detected by Western blot. Results:After co-transfection of miR-431-5p mimic and wild-type Smad4 reporter gene vector, luciferase activity of DPSC was significantly reduced (P<0.01), while which was not significantly changed after co-transfection of miR-431-5p mimic and mutant Smad4 reporter gene vector (P>0.05). Compared with mimic-NC group, expression of miR-431-5p was up-regulated, expression of Smad4 was down-regulated, ability of cells proliferation was increased, staining of calcified nodules was light, ALP activity was reduced, and expression of OCN protein was down-regulated in miR-431-5p mimic group (P<0.05). Compared with mimic-NC group, ability of cells proliferation was decreased, staining of calcified nodules was deepen, ALP activity was increased, and the expression of OCN was up-regulated in pc-Smad4 group (P<0.05). Compared with pc-Smad4 group, ability of cells proliferation was significantly increased, staining of calcified nodules was light, ALP activity was decreased, and the expression of OCN protein was down-regulated in pc-Smad4+miR-431-5p mimic group (P<0.05). Conclusions:The miR-431-5p can inhibit osteogenic differentiation of DPSC and promote their proliferation by inhibiting the expression of Smad4 protein.

Key words: dental pulp stem cell, miR-431-5p, Smad4, proliferation, osteogenic differentiation

References 13

[1]	赵振宇, 李影, 王港, 等. p38丝裂原活化蛋白激酶对人乳牙牙髓干细胞成骨分化能力的影响[J]. 中华老年口腔医学杂志, 2019, 17(3):7-12.
[2]	孙帅, 李军. 骨形态发生蛋白调控牙髓干细胞成牙本质分化的研究进展[J]. 中华实用诊断与治疗杂志, 2020, 34(11):107-110.
[3]	Lu X , Chen X , Xing J , et al. miR-140-5p regulates the odontoblastic differentiation of dental pulp stem cells via the Wnt1/β-catenin signaling pathway[J]. Stem Cell Res Ther, 2019, 10(1):226.
[4]	Sun DG, Xin BC, Wu D, et al. miR‐140‐5p‐mediated regulation of the proliferation and differentiation of human dental pulp stem cells occurs through the lipopolysaccharide/toll‐like receptor 4 signaling pathway[J]. Eur J Oral Sci, 2017, 125(6):419-425.
[5]	Qiu Z , Lin S , Hu X , et al. Involvement of miR‐146a‐5p/neurogenic locus notch homolog protein 1 in the proliferation and differentiation of STRO‐1+ human dental pulp stem cells[J]. Eur J Oral Sci, 2019, 127(4):294-303.
[6]	吴韫慧, 郭磊, 蒋翠霞,等. 人牙髓干细胞增殖分化过程中的miR-431[J]. 中国组织工程研究, 2019, 23(21):49-54.
[7]	Xu CM , Chen LX , Gao F , et al. MiR-431 suppresses proliferation and metastasis of lung cancer via down-regulating DDX5[J]. Eur Rev Med Pharmacol Sci, 2019, 23(2):699-707.
[8]	Wang Y , Zhang K , Yuan X , et al. miR-431-5p regulates cell proliferation and apoptosis in fibroblast-like synoviocytes in rheumatoid arthritis by targeting XIAP[J]. Arthritis Res Ther, 2020, 22(1):231.
[9]	姜力铭, 宋戈, 夏商, 等. 转化生长因子β1对人牙髓干细胞成骨分化作用研究[J]. 中国实用口腔科杂志, 2018, 11(9):4-7.
[10]	刘亚楠, 李祥伟, 孙宏晨,等. bFGF调控DPSCs迁移,增殖和分化的研究进展[J]. 口腔医学, 2017, 37(8):746-750.
[11]	李鹏, 张晓莉, 艾力麦尔丹·艾尼瓦尔,等. 骨形成蛋白-2与转化生长因子-β3对牙髓干细胞成骨向分化的对比研究[J]. 中华实用诊断与治疗杂志, 2019, 33(1):7-10.
[12]	Hu X, Zhong Y, Kong Y, et al. Lineage-specific exosomes promote the odontogenic differentiation of human dental pulp stem cells (DPSCs) through TGFβ1/smads signaling pathway via transfer of microRNAs[J]. Stem Cell Res Ther, 2019, 10(1):170.
[13]	木合塔尔·霍加. 转化生长因子β3促进兔牙髓干细胞成骨向分化机制的初步研究[J]. 口腔医学, 2019, 39(2):97-102.

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[5]	Yong HU. Effects of macrophages induced by Fusobacterium nucleatum on proliferation and migration of tongue squamous cell carcinoma Cal-27 cells [J]. Oral Biomedicine, 2021, 12(3): 166-169.
[6]	Xinquan 无Jiang. Magnesium-doped TiO2 coating on titanium by miro-arc oxidation promoting osteogenic differentiation of rat BMSCs [J]. Oral Biomedicine, 2021, 12(2): 76-81.
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[8]	. In vitro evaluation of biological activity on titanium dioxide nanotubes loaded with chitosan hydrogel crosslinked with genipin [J]. , 2020, 11(3): 164-167.
[9]	. The regulation of miR-21 in inflammatory response and osteogenic differentiation of periodontal ligament stem cells [J]. , 2020, 11(2): 86-91.
[10]	. The Influence of inflammatory micro-environment on regenerative capacity of PDLSCs and UCMSCs [J]. , 2020, 11(2): 71-75.
[11]	. The effect of stiffness of matrix on the osteoblasts growth and differentiation [J]. , 2020, 11(1): 13-18.
[12]	. Effects of miR-9 on the function of human periodontal ligament stem cells [J]. , 2019, 10(3): 114-117.
[13]	. Effect of electrospun SF/CS composite fiber scaffold on cell proliferation and osteogenic differentiation of hBMSCs in vitro [J]. , 2019, 10(2): 73-82.
[14]	. Preparation and biocompatibility of polycaprolactone/type I collagen/fluoroapatite composite scaffold [J]. , 2019, 10(1): 12-16.
[15]	Hao-Qing YANG. SFRP2 promoted osteogenic differentiation of adipose-derived stem cells [J]. , 2018, 9(3): 120-124.

Experimental study about the effects of miR-431-5p on the proliferation and differentiation of human dental pulp stem cells by targeting Smad4

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