[Effects of let-7a on proliferation, osteogenic differentiation and apoptosis of human dental pulp stem cells].

Q4 Medicine 上海口腔医学 Pub Date : 2023-10-01
Yan Liu, Dan-Ni Shan, Jing Sun, Yu-Xi Zou, Chang-Yong Yuan
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引用次数: 0

Abstract

Purpose: To study the effect and possible mechanism of let-7a on proliferation, differentiation and apoptosis of human dental pulp stem cell (hDPSCs).

Methods: The cells were divided into four groups: overexpression control (let-7a control/let-7a agomir control), overexpression let-7a (let-7a mimics/let-7a agomir), knockdown let-7a control (let-7a inhibitor control) and knockdown let-7a (let-7a inhibitor). Cell counting kit-8 assay(CCK-8) was used to detect the proliferation of cells at 24 hours, 48 hours and 72 hours after transfection. Calcified nodules were detected by Alizarin red staining. The protein expression of alkaline phosphatase (ALP), osteopontin (OPN), 4E-binding protein 1 (4EBP1), p-4EBP1, mammalian target of rapamycin (mTOR) and p-mTOR were detected by Western blot. Annexin V-APC/7-AAD cell apoptosis detection kit was used to detect the level of apoptosis after transfection. Statistical analysis was performed using GraphPad Prism 5.0 software.

Results: Let-7a inhibited proliferation of hDPSCs and promoted odontoblast differentiation and apoptosis. Let-7a down-regulated the expression of 4EBP1, p-4EBP1, mTOR and p-mTOR.

Conclusions: Let-7a may inhibit proliferation of hDPSCs and promote their differentiation and apoptosis by inhibiting mTOR-4EBP1 molecular pathway.

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[let-7a对人牙髓干细胞增殖、成骨分化和凋亡的影响]
目的:研究let-7a对人牙髓干细胞(hDPSCs)增殖、分化和凋亡的影响及可能机制:将细胞分为四组:过表达对照组(let-7a对照组/let-7a激动剂对照组)、过表达let-7a组(let-7a模拟组/let-7a激动剂组)、敲除let-7a对照组(let-7a抑制剂对照组)和敲除let-7a组(let-7a抑制剂组)。细胞计数试剂盒-8(CCK-8)用于检测转染后 24 小时、48 小时和 72 小时的细胞增殖情况。茜素红染色法检测钙化结节。通过 Western 印迹检测碱性磷酸酶(ALP)、骨生成素(OPN)、4E 结合蛋白 1(4EBP1)、p-4EBP1、哺乳动物雷帕霉素靶蛋白(mTOR)和 p-mTOR 的蛋白表达。Annexin V-APC/7-AAD 细胞凋亡检测试剂盒用于检测转染后的细胞凋亡水平。使用 GraphPad Prism 5.0 软件进行统计分析:结果:Let-7a抑制了hDPSCs的增殖,促进了颌骨母细胞的分化和凋亡。Let-7a下调了4EBP1、p-4EBP1、mTOR和p-mTOR的表达:结论:Let-7a可通过抑制mTOR-4EBP1分子通路来抑制hDPSCs的增殖并促进其分化和凋亡。
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来源期刊
上海口腔医学
上海口腔医学 Medicine-Medicine (all)
CiteScore
0.30
自引率
0.00%
发文量
5299
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