骨髓间充质干细胞对间充质样成釉细胞瘤细胞功能影响的研究  

Effects of BMSC on promoting M-AMC proliferation, migration and invasion

作　　者：肖语欢 李瑾 周薇娜[1] 江飞[2] 张娟 XIAO Yuhuan;LI Jin;ZHOU Weina;JIANG Fei;ZHANG Juan(Department of Temporomandibular Joint,The Affiliated Stomatological Hospital of Nanjing Medical University,Jiangsu Province Key Laboratory of Oral Diseases,Jiangsu Province Engineering Research Center of Stomatological Translational Medicine,Nanjing 210029,China;Department of General Dentistry,The Affiliated Stomatological Hospital of Nanjing Medical University,Nanjing 210029,China;Zhenjiang Stomatological Hospital Dantu Branch,Zhenjiang 212000,China)

机构地区：[1]南京医科大学附属口腔医院颞颌关节病科,江苏省口腔疾病研究重点实验室,江苏省口腔转化医学工程研究中心,江苏南京210029 [2]南京医科大学附属口腔医院综合诊疗科,江苏南京210029 [3]镇江市口腔医院丹徒分院,江苏镇江212000

出　　处：《口腔生物医学》2023年第1期6-11,共6页Oral Biomedicine

基　　金：国家自然科学基金(81900960);镇江市重点研发计划(社会发展)项目(SH2020070)。

摘　　要：目的:研究骨髓间充质干细胞(BMSC),对间充质样成釉细胞瘤细胞(M-AMC)的增殖、侵袭、迁移以及自噬水平的影响,初步探讨BMSC对M-AMC功能影响的部分作用和机制。方法:组织块结合酶消化法分离培养M-AMC并鉴定,构建非接触M-AMC/BMSC共培养体系,通过EdU检测M-AMC增殖变化,利用结晶紫染色、Western blot等检测M-AMC迁移、侵袭功能和自噬相关蛋白的改变。结果:通过原代分离培养得到的M-AMC,主要来源于间质,并具有一定的干性和自我更新能力。共培养组3、7 d M-AMC增殖的细胞数量明显高于对照组(P<0.01),共培养组在24、48、72 h的Transwell迁移和侵袭出的细胞数量均高于对照组(P<0.01),M-AMC在和BMSC共培养后,自噬表达水平显著增强(P<0.01)。结论:BMSC的旁分泌作用可以上调M-AMC自噬水平,从而促进M-AMC增殖、迁移、侵袭能力。Objective:To explore whether bone-marrow mesenchymalstem cell(BMSC)could affect ameloblastoma development and recurrence by promoting the proliferation,migration,invasion of mesenchymal cell-like ameloblastoma cell(M-AMC)via up-regulating their autophagy level.Methods:M-AMC was isolated and identified through tissue pieces and enzymatic digestion culture.The source of M-AMC was identified by Western blot.In vitro cell cloning experiments and flow experiments were also used to determine the expression level of M-AMC self-renewal ability.The migration and invasion ability of M-AMC at 24,48 and 72 h were determined by transwell and laying matrigel matrix gel experiments,and the invasion levels of M-AMC and autophagy were measured by Western blot.Results:In the M-AMC obtained by primary isolation cultures,the interstitial-derived M-AMC accounts for the main population,and has certain stemness and self-renewal capacity.The number of cells proliferating from co-culture group was significantly higher than control group on day 3 and day 7(P<0.01).The number of M-AMC migrated and invaded at the three time points of 24,48 and 72 h significantly higher than that of the control group(P<0.01).Invasion ability and autophagy level of M-AMC expression were significantly enhanced after coculture with BMSC(P<0.01).Conclusions:The paracrine effect of BMSC could promote the proliferation,migration and invasion of M-AMC,and these phenotypes were closely related to the up-regulated autophagy in M-AMC.

关 键 词：间充质样成釉细胞瘤细胞 骨髓间充质干细胞 自噬 共培养 

分 类 号：R780.2[医药卫生—口腔医学]