AFF4在年轻恒牙牙髓炎中的表达及对牙髓细胞分化的影响-东南大学学报医学版

AFF4在年轻恒牙牙髓炎中的表达及对牙髓细胞分化的影响

单位：新疆医科大学第五附属医院口腔科, 新疆乌鲁木齐 830011

分类号：R781.31

出版年·卷·期（页码）：2022·41·第四期（517-524）

摘要：

目的：探讨AF4/FMR2家族成员4（AFF4）在年轻恒牙牙髓炎中的表达特征和对牙髓细胞（DPCs）成骨分化的影响。方法：荧光定量PCR （qRT-PCR）、蛋白质印迹法、免疫组织化学法检测AFF4的表达。培养DPCs，用AFF4过表达载体（pcDNA-AFF4组）和过表达载体对照（pcDNA-ctrl组）转染DPCs，CCK-8法检测细胞增殖，Annexin V-FITC/PI双染法检测细胞的凋亡，碱性磷酸酶（ALP）活性检测和茜素红S （ARS）染色分析细胞的成骨分化。qRT-PCR检测分化标志物骨钙素（OCN）、骨桥蛋白（OPN）、骨形成锌指转录因子（OSX）、肌节同源框基因2（MSX2）、远端同源框基因5（DLX5）和runt相关转录因子2（RUNX2）的表达。结果：年轻恒牙牙髓炎中AFF4表达下调（P<0.05）。与pcDNA-ctrl组比，pcDNA-AFF4组中细胞增殖率、ALP活性、矿化结构都增加，而凋亡率降低，且OCN、OPN、OSX、MSX2、DLX5、RUNX2的表达上调（均P<0.05）。结论：年轻恒牙牙髓炎中AFF4表达降低，过表达AFF4促进DPCs增殖和成骨分化，并抑制细胞凋亡。

Objective: To investigate the expression characteristics of AF4/FMR2 family member 4 (AFF4) in young permanent teeth pulpitis and its effect on the osteogenic differentiation of dental pulp cells(DPCs). Methods: Real-time quantitative PCR (qRT-PCR), Western blot and immunohistochemistry were used to detect the expression of AFF4. Human DPCs were cultured and transfected with AFF4 overexpression vector (pcDNA-AFF4 group) and overexpression vector control (pcDNA-ctrl group). Cell proliferation was detected by CCK-8 assay. The apoptosis of cells was detected by Annexin V-FITC/PI double staining. The osteogenic differentiation of cells was analyzed by alkaline phosphatase (ALP) activity detection and Alizarin red S staining. The expression of differentiation markers osteocalcin (OCN), osteopontin (OPN), osterix (OSX), Msh Homeobox 2 (MSX2), distal-less homeobox 5 (DLX5) and runt-related transcription factor 2 (RUNX2) were detected by qRT-PCR. Results: AFF4 was down-regulated in young permanent tooth pulpitis (P<0.05). Compared with the pcDNA-ctrl group, the cell proliferation rate, ALP activity, and mineralization structure in the pcDNA-AFF4 group increased, while the apoptosis rate decreased, and the expressions of OCN, OPN, OSX, MSX2, DLX5, and RUNX2 were all up-regulated (all P <0.05). Conclusion: The expression of AFF4 is decreased in young permanent teeth pulpitis. Overexpression of AFF4 promotes the proliferation and osteogenic differentiation of DPCs, and inhibits cell apoptosis.

参考文献：

[1] TOMASELLO L, MAUCERI R, COPPOLA A, et al.Mesenchymal stem cells derived from inflamed dental pulpal and gingival tissue:a potential application for bone formation[J].Stem Cell Res Ther, 2017, 8(1):179-189.
[2] NUTI N, CORALLO C, CHAN B M, et al.Multipotent differentiation of human dental pulp stem cells:a literature review[J].Stem Cell Rev Rep, 2016, 12(5):511-523.
[3] TAN Q, CAO Y, ZHENG X, et al.BMP4-regulated human dental pulp stromal cells promote pulp-like tissue regeneration in a decellularized dental pulp matrix scaffold[J].Odontology, 2021, 109(4):895-903.
[4] ZHOU C C, XIONG Q C, ZHU X X, et al.AFF1 and AFF4 differentially regulate the osteogenic differentiation of human MSCs[J].Bone Res, 2017, 5(1):17044-17055.
[5] GUO Y C, WANG M Y, ZHANG S W, et al.Ubiquitin-specific protease USP34 controls osteogenic differentiation and bone formation by regulating BMP2 signaling[J].EMBO J, 2018, 37(20):e99398-e99409.
[6] CARRIZALES-SEPÚLVEDA E F, ORDAZ-FARÍAS A, VERA-PINEDA R, et al.Periodontal disease, systemic inflammation and the risk of cardiovascular disease[J].Heart Lung Circ, 2018 (11):1327-1334.
[7] CASAGRANDE L, CORDEIRO M M, NÖR S A, et al.Dental pulp stem cells in regenerative dentistry[J].Odontology, 2011, 99(1):e7-e19.
[8] MANJI F, DAHLEN G, FEJERSKOV O.Caries and periodontitis:contesting the conventional wisdom on their aetiology[J].Caries Res, 2018, 52(1):548-564.
[9] KINANE D F, STATHOPOULOU P G, PAPAPANOU P N.Periodontal diseases[J].Nat Rev Dis Prim, 2017, 3(1):17038-17049.
[10] ANITUA E, TROYA M, ZALDUENDO M.Progress in the use of dental pulp stem cells in regenerative medicine[J].Cytotherapy, 2018, 20(4):479-498.
[11] HERNÁNDEZ-MONJARAZ B, SANTIAGO-OSORIO E, LEDESMA-MARTÍNEZ E, et al.Retrieval of a periodontally compromised tooth by allogeneic grafting of mesenchymal stem cells from dental pulp:a case report[J].J Int Med Res, 2018, 46 (1):2983-2993.
[12] BITOUN E, OLIVER P L, DAVIES K E.The mixed-lineage leukemia fusion partner AF4 stimulates RNA polymerase Ⅱ transcriptional elongation and mediates coordinated chromatin remodeling[J].Hum Mol Genet, 2007, 16(1):92-106.
[13] BISWAS D, MILNE T A, BASRUR V, et al.Function of leukemogenic mixed lineage leukemia 1 (MLL) fusion proteins through distinct partner protein complexes[J].Proc Natl, 2011, 108(1):15751-15756.
[14] CHEN Y, CRAMER P.Structure of the super-elongation complex subunit AFF4 Cterminal homology domain reveals requirements for AFF homo-and heterodimerization[J].J Biol Chem, 2019, 294(1):10663-10673.
[15] SCHULZE-GAHMEN U, LU H, ZHOU Q, et al.Alber, AFF4 binding to Tat-P-TEFb indirectly stimulates TAR recognition of super elongation complexes at the HIV promoter[J].Elife, 2014(3):e0237-e0248.
[16] XIAO Q, ZHANG Y, QI X, et al.AFF4 regulates osteogenic differentiation of human dental follicle cells[J].Int J Oral Sci, 2020, 12(1):20-29.
[17] CHEN Y, CRAMER P.Structure of the super-elongation complex subunit AFF4 C-terminal homology domain reveals requirements for AFF homo-and heterodimerization[J].J Biol Chem, 2019, 294(1):10663-10673.

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