Let-7c靶向IGF2BP2调控人牙髓干细胞增殖的分子机制

doi:10.19439/j.sjos.2024.06.002

上海口腔医学 ›› 2024, Vol. 33 ›› Issue (6): 572-579.doi: 10.19439/j.sjos.2024.06.002

Let-7c靶向IGF2BP2调控人牙髓干细胞增殖的分子机制

王雨珊^1*, 孙梦馨^1*, 杨屹城¹, 衡笑¹, 张雨欣¹, 杨建光^1,2#, 刘岩^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

收稿日期:2023-12-15 修回日期:2024-02-06 出版日期:2024-12-25 发布日期:2025-01-07
通讯作者: 刘岩,E-mail: 1563625177@qq.com;杨建光,E-mail: jian-guangyang@xzhmu.edu.cn。^#共同通信作者
作者简介:王雨珊（2003-）,女,本科,E-mail：2404320400@qq.com;孙梦馨（2003-）,女,本科,E-mail：207517306@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资助项目（22KJB320024）;徐州市重点研发计划项目（KC22218）;徐州市科技局重点研发计划（社会发展）（KC21225）

Molecular mechanisms of let-7c targeting IGF2BP2 to regulate the proliferation of human dental pulp stem cells

WANG Yu-shan¹, SUN Meng-xin¹, YANG Yi-cheng¹, HENG Xiao¹, ZHANG Yu-xin¹, YANG Jian-guang^1,2, LIU Yan^1,2

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

Received:2023-12-15 Revised:2024-02-06 Online:2024-12-25 Published:2025-01-07

摘要/Abstract

摘要： 目的: 探讨let-7c过表达对人类牙髓干细胞（human dental pulp stem cells,hDPSCs）增殖的影响及其作用机制。方法: 分离培养hDPSCs,应用let-7c模拟物和抑制剂转染hDPSCs。利用qRT-PCR检测转染效率,利用CCK-8检测细胞增殖情况。应用流式细胞术检测细胞周期,通过蛋白质印迹分析let-7c抑制细胞增殖的作用机制。应用小干扰RNA（small interfer RNA,siRNA）瞬时转染方法构建针对胰岛素样生长因子2 mRNA结合蛋白2 (insulin-like growth factor 2 mRNA binding protein 2,IGF2BP2)基因的细胞模型,通过生物信息学分析和双荧光素酶报告基因实验探讨分析let-7c是否直接靶向IGF2BP2。采用SPSS 17.0软件包对数据进行统计学分析。结果: Let-7c过表达抑制细胞增殖。Let-7c通过调节S和G2/M周期阻断细胞增殖,直接与hDPSCs中与细胞周期调控有关的一组信使RNA（messenger RNA,mRNA）序列IGF2BP2结合,靶向抑制IGF2BP2的表达,两者的表达水平呈现负相关关系。结论: Let-7c通过靶向抑制IGF2BP2表达,抑制hDPSCs增殖。

关键词: Let-7c, IGF2BP2, 人类牙髓干细胞, 细胞增殖, 细胞周期

Abstract: PURPOSE: To investigate the effect and mechanism of let-7c overexpression on the proliferation of human dental pulp stem cells (hDPSCs). METHODS: hDPSCs were isolated and cultured. After transfected with let-7c mimics and inhibitors, transfection efficiency was detected by qRT-PCR, and cell proliferation was detected by cell counting kit-8 (CCK-8). Flow cytometry was applied to determine the cell cycle. The mechanism of let-7c inhibiting cell proliferation was analyzed by Western blot. A cell model targeting insulin-like growth factor 2 mRNA binding protein 2 (IGF2BP2) was constructed by transient transfection of small interfering RNA(siRNA) and analyzed by bioinformatics and dual luciferase reporter. Bioinformatics analysis and dual luciferase reporter gene assay were performed to investigate whether let-7c directly targeted IGF2BP2. The data were statistically analyzed using SPSS 17.0 software package. RESULTS: Overexpression of let-7c inhibited cell proliferation. Let-7c blocked cell proliferation by regulating the S and G2/M cycles, and directly bound to a group of messenger RNA (mRNA) sequences related to cell cycle regulation, IGF2BP2, and targeting and inhibiting IGF2BP2 expression, with a negative correlation between the two expression levels. CONCLUSIONS: let-7c inhibites the proliferation of hDPSCs by targeting and inhibiting IGF2BP2 expression.

Key words: Let-7c, IGF2BP2, HDPSCs, Cell proliferation, Cell cycle

中图分类号:

R781.3

王雨珊, 孙梦馨, 杨屹城, 衡笑, 张雨欣, 杨建光, 刘岩. Let-7c靶向IGF2BP2调控人牙髓干细胞增殖的分子机制[J]. 上海口腔医学, 2024, 33(6): 572-579.

WANG Yu-shan, SUN Meng-xin, YANG Yi-cheng, HENG Xiao, ZHANG Yu-xin, YANG Jian-guang, LIU Yan. Molecular mechanisms of let-7c targeting IGF2BP2 to regulate the proliferation of human dental pulp stem cells[J]. Shanghai Journal of Stomatology, 2024, 33(6): 572-579.

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Let-7c靶向IGF2BP2调控人牙髓干细胞增殖的分子机制

Molecular mechanisms of let-7c targeting IGF2BP2 to regulate the proliferation of human dental pulp stem cells

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