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间充质基质细胞分泌组用于胶质母细胞瘤治疗的安全性分析。

Analysis of the safety of mesenchymal stromal cells secretome for glioblastoma treatment.

作者信息

Onzi Giovana Ravizzoni, Ledur Pítia Flores, Hainzenreder Luana Dimer, Bertoni Ana Paula Santin, Silva Andrew Oliveira, Lenz Guido, Wink Márcia Rosângela

机构信息

Laboratory of Cell Biology, Universidade Federal de Ciências da Saúde de Porto Alegre, Porto Alegre, Brazil; Department of Biophysics and Center of Biotechnology, Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, RS, Brazil.

Department of Biophysics and Center of Biotechnology, Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, RS, Brazil.

出版信息

Cytotherapy. 2016 Jul;18(7):828-37. doi: 10.1016/j.jcyt.2016.03.299. Epub 2016 May 17.

Abstract

BACKGROUND AIMS

The purpose of this study was to investigate whether the secretome of human adipose-derived stem cells (hADSC) affects human glioblastoma (GBM) cancer stem cell (CSC) subpopulation or has any influence on drug resistance and cell migration, evaluating the safety of hADSCs for novel cancer therapies.

METHODS

hADSCs were maintained in contact with fresh culture medium to produce hADSCs conditioned medium (CM). GBM U87 cells were cultured with CM and sphere formation, expression of genes related to resistance and CSCs-MGMT, OCT4, SOX2, NOTCH1, MSI1-and protein expression of OCT4 and Nanog were analyzed. The influence of hADSC CM on GBM resistance to temozolomide (TMZ) was evaluated by measuring cumulative population doubling and hADSC CM influence on tumor cell migration was analyzed using transwell assay.

RESULTS

hADSC CM did not alter CSC-related features such as sphere-forming capacity and expression of genes related to CSC. hADSC CM treatment alone did not change proliferation rate of U87 cells and, most important, did not alter the response of tumor cells to TMZ. However, hADSC CM secretome increased the migration capacity of glioblastoma cells.

DISCUSSION

hADSC CM neither induced an enrichment of CSCs in U87 cells population nor interfered in the response to TMZ in culture. Nevertheless, paracrine factors released by hADSCs were able to modulate glioblastoma cells migration. These findings provide novel information regarding the safety of using hADSCs against cancer and highlight the importance of considering hADSC-tumor cells interactions in tumor microenvironment in the design of novel cell therapies.

摘要

背景目的

本研究旨在调查人脂肪来源干细胞(hADSC)的分泌组是否会影响人胶质母细胞瘤(GBM)癌干细胞(CSC)亚群,或对耐药性和细胞迁移有任何影响,评估hADSC在新型癌症治疗中的安全性。

方法

将hADSC与新鲜培养基接触培养以产生hADSC条件培养基(CM)。用CM培养GBM U87细胞,分析成球情况、与耐药相关基因及CSC相关基因-MGMT、OCT4、SOX2、NOTCH1、MSI1的表达,以及OCT4和Nanog的蛋白表达。通过测量累积群体倍增来评估hADSC CM对GBM对替莫唑胺(TMZ)耐药性的影响,并使用Transwell实验分析hADSC CM对肿瘤细胞迁移的影响。

结果

hADSC CM未改变与CSC相关的特征,如成球能力和与CSC相关基因的表达。单独的hADSC CM处理未改变U87细胞的增殖率,最重要的是,未改变肿瘤细胞对TMZ的反应。然而,hADSC CM分泌组增加了胶质母细胞瘤细胞的迁移能力。

讨论

hADSC CM既未诱导U87细胞群体中CSC富集,也未干扰培养中对TMZ的反应。然而,hADSC释放的旁分泌因子能够调节胶质母细胞瘤细胞的迁移。这些发现提供了关于使用hADSC抗癌安全性的新信息,并强调了在设计新型细胞疗法时考虑肿瘤微环境中hADSC与肿瘤细胞相互作用的重要性。

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