成釉细胞瘤细胞抑制骨髓间充质干细胞成骨分化的研究

›› 2020, Vol. 40 ›› Issue (12): 1067-1072.

成釉细胞瘤细胞抑制骨髓间充质干细胞成骨分化的研究

雷港¹,武和明²,江飞¹

1. 南京医科大学附属口腔医院
2. 南京医科大学附属口腔医院口腔颌面外科

收稿日期:2020-05-21 修回日期:2020-06-28 出版日期:2020-12-28 发布日期:2020-12-22
通讯作者: 江飞 E-mail:nykqjf@njmu.edu.cn
基金资助:
江苏省高校自然科学研究面上项目;江苏省高校优势学科建设工程资助项目;国家自然科学基金

Research on human amelobastoma cells inhibiting the osteogenic differentiation of hBMSCs

Received:2020-05-21 Revised:2020-06-28 Online:2020-12-28 Published:2020-12-22

摘要/Abstract

摘要： 摘要：目的：研究成釉细胞瘤细胞（human ameloblastoma-derived cells, hAMCs）对骨髓间充质干细胞（human bone marrow-derived mesenchymal stem cells, hBMSCs）增殖、成骨分化，以及自噬水平的影响，初步探讨成釉细胞瘤抑制骨髓干细胞骨再生的部分机制。方法：组织块法结合酶消化法分离培养hAMCs并鉴定，收集其条件培养液（hAMC conditional media, hAMC-CM）。以hAMC-CM刺激hBMSCs，通过CCK-8检测hBMSCs增殖变化，碱性磷酸酶实验，茜素红染色及Western blot检测hBMSCs成骨分化相关表达的变化，Western blot 检测hBMSCs自噬相关指标的变化。结果： hAMC-CM处理hBMSCs后，细胞早期及晚期成骨相关指标下调。同时自噬相关蛋白表达下调。结论：hAMCs可通过旁分泌作用抑制hBMSCs自噬水平，进而抑制其成骨分化。

关键词: 成釉细胞瘤细胞, 骨髓间充质干细胞, 骨再生, 自噬

Abstract: Abstract: Object: To explore the mechanism of ameloblastoma in inhibiting proliferation, osteogenic differentiation and autophagy level of human bone marrow derived mesenchymal stem cells (hBMSCs) to affect bone regeneration. Method: Human ameloblastoma cells (hAMCs) were isolated and identified. The conditional media was collected to culture the hBMSCs. CCK-8 kit was used to detect the hBMSC proliferation, while ALP staining, Alizarin Red staining and Western-blot were applied to investigate the osteogenic differentiation and autophagy status of the hBMSCs. Results: The osteogenic phenotype of hBMSCs were inhibited by hAMC-CM. The markers of osteogenesis and autophagy in hBMSCs were down-regulated, such as Runx2, Osx, ALP, OCN, OPN, Collagen I, ATG5, Beclin and LC3b. Conclusion: hAMCs could affect autophagy status of hBMSCs via their paracrine functions to inhibit osteogenesis.

Key words: hAMCs, bone marrow mesenchymal stem cells, bone regeneration, autophagy

中图分类号:

R349.5

雷港武和明江飞. 成釉细胞瘤细胞抑制骨髓间充质干细胞成骨分化的研究[J]. 口腔医学, 2020, 40(12): 1067-1072.

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