Gu Yingzhi, Bai Yuxing
Department of Orthodontics, Beijing Stomatological Hospital, Capital Medical University, Beijing, China.
Oral Dis. 2024 May;30(4):2398-2409. doi: 10.1111/odi.14705. Epub 2023 Aug 3.
Promoting the osteogenic differentiation of periodontal ligament stem cells (PDLSCs) is a way to regenerate periodontal bone. This study aimed to determine whether lncRNA MALAT1 promotes the osteogenic differentiation of human PDLSCs in vitro.
Human PDLSCs were extracted from the human periodontal ligament, and after osteogenic differentiation was induced using osteogenic medium, the human PDLSCs were transfected with siRNA-MALAT1, miR-93-5p mimics, and miR-93-5p inhibitors. The expression of osteogenesis-related genes was assessed by RT-qPCR and western blotting, alkaline phosphatase (ALP) activity was assessed by ALP activity assay, and the formation of mineralized nodules was assessed by alizarin red S (ARS) staining. RNA immunoprecipitation (RIP) and luciferase assays were performed to assess the binding of MALAT1, miR-93-5p, and SMAD5.
The expression of lncRNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) was upregulated, while that of miR-93-5p was downregulated after PDLSC osteogenic differentiation. Knockdown of MALAT1 inhibited the osteogenic differentiation of PDLSCs, and MALAT1 expression negatively correlated with miR-93-5p expression. miR-93-5p inhibited the osteogenic differentiation of human PDLSCs by specifically binding to SMAD5.
MALAT1 regulates human PDLSC differentiation by regulating the miR-93-5p/SMAD5 axis.
促进牙周膜干细胞(PDLSCs)的成骨分化是牙周骨再生的一种方式。本研究旨在确定长链非编码RNA MALAT1是否在体外促进人PDLSCs的成骨分化。
从人牙周膜中提取人PDLSCs,使用成骨培养基诱导成骨分化后,将人PDLSCs用siRNA-MALAT1、miR-93-5p模拟物和miR-93-5p抑制剂进行转染。通过RT-qPCR和蛋白质免疫印迹法评估成骨相关基因的表达,通过碱性磷酸酶(ALP)活性测定评估ALP活性,通过茜素红S(ARS)染色评估矿化结节的形成。进行RNA免疫沉淀(RIP)和荧光素酶测定以评估MALAT1、miR-93-5p和SMAD5的结合情况。
PDLSCs成骨分化后,长链非编码RNA转移相关肺腺癌转录本1(MALAT1)的表达上调,而miR-93-5p的表达下调。敲低MALAT1抑制了PDLSCs的成骨分化,且MALAT1表达与miR-93-5p表达呈负相关。miR-93-5p通过特异性结合SMAD5抑制人PDLSCs的成骨分化。
MALAT1通过调节miR-93-5p/SMAD5轴调控人PDLSCs的分化。