Let-7a对人牙髓干细胞增殖、凋亡和成骨分化的影响

doi:10.19439/j.sjos.2023.05.004

上海口腔医学 ›› 2023, Vol. 32 ›› Issue (5): 468-474.doi: 10.19439/j.sjos.2023.05.004

Let-7a对人牙髓干细胞增殖、凋亡和成骨分化的影响

刘岩^1,2,*, 单丹妮^3,*, 孙晶¹, 邹雨希¹, 袁长永^1,2

1.徐州医科大学,江苏徐州 221004;
2.徐州医科大学附属口腔医院,江苏徐州 221002;
3.南京大学医学院附属口腔医院,南京市口腔医院,江苏南京 210008

收稿日期:2022-04-26 修回日期:2022-07-08 出版日期:2023-10-25 发布日期:2023-11-03
通讯作者: 袁长永,E-mail：yuanchangyong1983@foxmail.com
作者简介:刘岩（1989-）,女,博士,讲师,E-mail：1563625177@qq.com;单丹妮（2000-）,女,在读硕士研究生,E-mail: 2811108118@qq.com。^*并列第一作者
基金资助:
江苏省高等学校基础科学研究面上省资助项目（22KJB320024）; 徐州市重点研发计划项目（KC22218）

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

LIU Yan^1,2, SHAN Dan-ni³, SUN Jing¹, ZOU Yu-xi¹, YUAN Chang-yong^1,2

1. Xuzhou Medical University. Xuzhou 221004;
2. Affiliated Stomatological Hospital of Xuzhou Medical University. Xuzhou 221002;
3. Nanjing Stomatological Hospital, Affiliated Hospital of Medical School, Nanjing University. Nanjing 210008, Jiangsu Province, China

Received:2022-04-26 Revised:2022-07-08 Online:2023-10-25 Published:2023-11-03

摘要/Abstract

摘要： 目的：研究let-7a对人牙髓干细胞（human dental pulp stem cells,hDPSCs）增殖、成骨分化、凋亡的影响及可能的作用机制。方法：通过let-7a mimics、let-7a agomir及let-7a inhibitor转染hDPSCs。将细胞分为4组,过表达对照组（let-7a control组/let-7a agomir control 组）、过表达let-7a组（let-7a mimics组/let-7a agomir 组）、敲低let-7a对照组（let-7a inhibitor control组）和敲低let-7a组（let-7a inhibitor组）。CCK-8法检测细胞在转染后继续培养24、48、72 h的增殖情况,茜素红染色检测钙化结节。Western印迹法检测细胞碱性磷酸酶（alkaline phosphatase,ALP）、骨桥蛋白（osteopontin,OPN）、4E-结合蛋白（4E-binding protein 1,4EBP1）、p-4EBP1、哺乳动物类雷帕霉素靶蛋白（mammalian target of rapamycin,mTOR）和p-mTOR蛋白表达。Annexin V-APC/ 7-AAD细胞凋亡检测试剂盒检测转染后的细胞凋亡水平。采用GraphPad Prism 5.0软件进行统计分析。结果：Let-7a抑制hDPSCs增殖,对细胞成骨向分化和凋亡具有促进作用。Let-7a可下调细胞内4EBP1、p-4EBP1、mTOR和p-mTOR的表达水平。结论：Let-7a可能通过抑制mTOR-4EBP1分子通路,从而抑制hDPSCs增殖,促进其成骨分化和凋亡。

关键词: let-7a, 人牙髓干细胞, 增殖, 分化, 凋亡

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.

Key words: Let-7a, Human dental pulp stem cells, Proliferation, Differentiation, Apoptosis

中图分类号:

R781.3

刘岩, 单丹妮, 孙晶, 邹雨希, 袁长永. Let-7a对人牙髓干细胞增殖、凋亡和成骨分化的影响[J]. 上海口腔医学, 2023, 32(5): 468-474.

LIU Yan, SHAN Dan-ni, SUN Jing, ZOU Yu-xi, YUAN Chang-yong. Effects of let-7a on proliferation, osteogenic differentiation and apoptosis of human dental pulp stem cells[J]. Shanghai Journal of Stomatology, 2023, 32(5): 468-474.

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Let-7a对人牙髓干细胞增殖、凋亡和成骨分化的影响

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

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