PVT1促进在胶质瘤C6细胞微环境中的BMSCs增殖和迁移  

PVT1 Promoted the Proliferation and Migration Ability of BMSCs in Glioma C6 Microenvironment

作　　者：刘东蓉 刘艳 吴培连 郑雷蕾[1,2,3] 黄小亚 孟雪欢 胡赟 LIU Dongrong;LIU Yan;WU Pei-lian;ZHENG Lei-lei;HUANG Xiao-ya;MENG Xue-huan;HU Yun(The Affiliated Stomatology Hospital,Chongqing Medical University,Chongqing 401145,China;Chongqing Key Laboratory of Oral Diseases and Biomedical Sciences,Chongqing 401145,China;Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education,Chongqing 401145,China;The Second Affiliated Hospital,Chongqing Medical University,Chongqing 400010,China)

机构地区：[1]重庆医科大学附属口腔医院,重庆401145 [2]口腔疾病与生物医学重庆市重点实验室,重庆401145 [3]重庆市高校市级口腔生物医学工程重点实验室,重庆401145 [4]重庆医科大学附属第二医院,重庆400010

出　　处：《四川大学学报（医学版）》2021年第3期438-444,共7页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金面上项目(No.81470772);重庆市科卫联合医学科研重点项目(No.2018ZDXM020)资助。

摘　　要：目的探讨与胶质瘤C6细胞间接共培养后骨髓间充质干细胞(BMSCs)增殖和迁移能力的变化以及长链非编码RNA(lncRNA)浆细胞瘤变异异位基因1(plasmacytoma variant translocation 1 gene,PVT1)在这种变化中的作用。方法分离、培养、鉴定BMSCs后,采用Transwell小室将生长状态良好的BMSCs与胶质瘤C6细胞间接共培养(共培养组),单独培养的正常BMSCs为对照组。CCK8及软琼脂克隆形成实验检测2组细胞的增殖能力,流式细胞术检测细胞周期,划痕实验及Transwell法检测细胞的迁移能力,实时定量PCR(qRT-PCR)检测PVT1基因的表达。在共培养组BMSCs中转染siPVT1(si-PVT1组)和si-NC(si-NC组),转染后重复上述检测,并增加qRT-PCR检测周期蛋白基因CyclinD1及迁移相关基因基质金属蛋白酶2、9(MMP2、MMP9)的表达。结果所用BMSCs具有成骨、成脂分化能力。与对照组相比,共培养组BMSCs体积变小,排列紊乱,细胞间接触抑制消失,增殖及迁移能力增强,S期及G2期细胞比例增加,PVT1表达增加(P<0.05)。与si-NC组比较,si-PVT1组抑制共培养BMSCs的增殖及迁移能力,G1期细胞比例增加,S期细胞比例减少,PVT1、CyclinD1及MMP2、MMP9 mRNA表达也降低(P<0.05)。结论胶质瘤C6细胞微环境中BMSCs增殖及迁移能力增强可能与PVT1高表达有关。Objective To investigate the changes in the proliferation and migration ability of bone marrow mesenchymal stem cells(BMSCs)after indirect co-culturing with glioma C6 cells,and to examine the role of plasmacytoma variant translocation 1 gene(PVT1),a long non-coding RNA(lncRNA),in these changes.Methods After separation,cultivation and identification of BMSCs,BMSCs of good growth condition were picked out and indirectly cocultured with glioma C6 cells in Transwell chambers.These cells are henceforth referred to as the co-culture group.Normal BMSCs cultured separately were the control group.CCK-8 and soft agar colony formation assay were used to examine the proliferation ability of the two groups of cells.Flow cytometry was used to examine the cell cycle.Wound healing assay and Transwell assay were used to explore the migration ability of the cells.Quantitative real-time PCR(qRTPCR)was used to examine the genetic expression level of PVT1 in the two groups.The above-mentioned tests were repeated after the co-cultured BMSCs were transfected with si-PVT1(si-PVT1 group)and si-NC(si-NC group).In addition,qRT-PCR was done to evaluate the expression of CyclinD1,a cell cycle protein gene,and matrix metalloproteinases 2 and 9(MMP2 and MMP9),the migration-related genes in the si-PVT1 and si-NC transfected cocultured BMSCs.Results The BMSCs used in the present study possess the capability of osteogeneic and adipogenic differentiation.Compared with the control group,the co-cultured BMSCs had smaller size,disorderly arrangement and the lack of intercellular contact inhibition.The proliferation and migration ability was significantly enhanced,the proportions of S and G2 phase cells greatly increased and the expression level of PVT1 was significantly up-regulated(P<0.05)in the co-cultured group in comparison with those of the control group.When compared with the si-NC group,the si-PVT1 group showed inhibited proliferation and migration ability of the co-cultured BMSCs;the percentage of G1 phase cells increased,while that of S phas

关 键 词：长链非编码RNA PVT1 骨髓间充质干细胞 间接共培养 增殖 迁移 

分 类 号：R739.41[医药卫生—肿瘤]