Yao Xuanxuan, Qin Ruoshan, Cui Ziwei, He Dengqi, Sun Xiaorong, Sun Yuefeng, He Xiangyi
School of Dentistry, Lanzhou University, Lanzhou, Gansu 730000, P.R. China.
Prosthodontic Department, Stomatological Hospital of Lanzhou City, Lanzhou, Gansu 730000, P.R. China.
Exp Ther Med. 2024 Dec 17;29(2):33. doi: 10.3892/etm.2024.12783. eCollection 2025 Feb.
A human periodontal ligament cell line stably overexpressing the granulin precursor gene () was established through lentiviral mediation to explore the effects of on the proliferation and osteogenic capacity of human periodontal ligament cells (hPDLCs). In the present study, a homologous recombinant lentiviral plasmid, pLV-GRN, was constructed and transfected into the hPDLCs. The expression levels of and progranulin were assessed using reverse transcription-quantitative (RT-q)PCR and western blotting and the effect of on the proliferative capacity was determined using the MTT assay. The osteogenic capacity of human periodontal cells overexpressing was evaluated. The results showed that successful construction and transfection of the homologous recombinant lentiviral plasmid pLV-GRN led to the development of a stable periodontal cell line overexpressing . The MTT assay results revealed an enhanced proliferative capacity in the pLV-GRN group compared with that in the hPDLCs and pLV-puro groups (P<0.05). Alizarin red staining and alkaline phosphatase (ALP) activity assays indicated a significantly increased osteogenic capacity in the pLV-GRN group compared with the hPDLCs and pLV-puro groups (P<0.01). RT-qPCR demonstrated strong expression of the osteogenic genes ALP, runt-related transcription factor 2 (Runx2) and osteopontin (OPN) in the periodontal cells of the pLV-GRN group (P<0.05), whereas western blotting results corroborated the high expression of the osteogenic genes Runx-2 and OPN in the periodontal cells of the pLV-GRN group (P<0.05). In summary, the overexpression of significantly enhanced the proliferation and osteogenic capacity of hPDLCs. These findings provide an experimental foundation for periodontal tissue regeneration.
通过慢病毒介导建立了稳定过表达颗粒蛋白前体基因（）的人牙周膜细胞系，以探讨其对人牙周膜细胞（hPDLCs）增殖和成骨能力的影响。在本研究中，构建了同源重组慢病毒质粒pLV-GRN并将其转染到hPDLCs中。使用逆转录定量（RT-q）PCR和蛋白质印迹法评估颗粒蛋白前体基因（）和前颗粒蛋白的表达水平，并使用MTT法测定其对增殖能力的影响。评估了过表达颗粒蛋白前体基因（）的人牙周细胞的成骨能力。结果表明，同源重组慢病毒质粒pLV-GRN的成功构建和转染导致了稳定的过表达颗粒蛋白前体基因（）的牙周细胞系的建立。MTT分析结果显示，与hPDLCs组和pLV-puro组相比，pLV-GRN组的增殖能力增强（P<0.05）。茜素红染色和碱性磷酸酶（ALP）活性测定表明，与hPDLCs组和pLV-puro组相比，pLV-GRN组的成骨能力显著增强（P<0.01）。RT-qPCR显示，pLV-GRN组牙周细胞中碱性磷酸酶（ALP）、 runt相关转录因子2（Runx2）和骨桥蛋白（OPN）等成骨基因表达强烈（P<0.05），而蛋白质印迹结果证实pLV-GRN组牙周细胞中Runx-2和OPN等成骨基因高表达（P<0.05）。总之，颗粒蛋白前体基因（）的过表达显著增强了hPDLCs的增殖和成骨能力。这些发现为牙周组织再生提供了实验基础。
