Rat bone marrow mesenchymal stem cells undergo malignant transformation via indirect co-cultured with tumour cells
Rat bone marrow mesenchymal stem cells undergo malignant transformation via indirect co-cultured with tumour cells
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大鼠骨髓间充质干细胞与肿瘤细胞间接共培养发生恶性转化
DOI：
10.1002/cbf.2844
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发表时间：
2012-12-01
影响因子：
3.6
通讯作者：
Zhu, Jing
中科院分区：
文献类型：
--
作者：
Liu, Jianping;Zhang, Yalan;Zhu, Jing
骨髓间充质干细胞（Mesenchymal stem cells，MSCs）在再生医学和组织工程中具有潜在的应用价值，同时也是肿瘤治疗的潜在载体。然而，在肿瘤中使用MSC的安全性尚不清楚。在此，我们分析了肿瘤微环境中MSC的恶性转化。将大鼠骨髓间充质干细胞与大鼠恶性胶质瘤C6细胞共同培养，不进行细胞直接接触。七点以后？30天后，使用流式细胞术、实时定量PCR、免疫荧光和染色体分析评估细胞的转化。此外，野生型（WT）p53，突变型p53和mdm2使用蛋白质印迹法测定。几乎所有的骨髓间充质干细胞成为恶性表型细胞，WT p53表达显著降低，突变型p53和mdm2表达增加，沿着非整倍体核型。为了评估体内成瘤性，将MSC与C6细胞间接共培养7？天皮下移植到免疫缺陷小鼠中。细胞发展成一个大的肿瘤在注射部位内8？周，全身症状包括恶病质和脊柱侧凸。病理学和细胞学分析显示低分化的多形性细胞具有密集的血管网，并侵袭邻近的肌肉。这些数据表明，当暴露于肿瘤微环境时，MSC变成恶性癌细胞，并表明从癌细胞释放的因子在MSC的恶性转化中具有关键作用。版权所有（c）2012约翰威利父子有限公司
Mesenchymal stem cells (MSCs) have potential applications in regenerative medicine and tissue engineering as well as being potential carriers for tumour therapy. However, the safety of using MSCs in tumours is unknown. Herein, we analyse malignant transformation of MSCs in the tumour microenvironment. Rat bone marrow MSCs were cultured with malignant rat glioma C6 cells without direct cellcell contact. After 7?days, the cells were assessed for transformation using flow cytometry, real-time quantitative PCR, immunofluorescence and chromosomal analysis. In addition, wild-type (WT) p53, mutant p53 and mdm2 was determined using Western blotting. Almost all MSCs became phenotypically malignant cells, with significantly decreased WT p53 expression and increased expression of mutant p53 and mdm2, along with an aneuploid karyotype. To evaluate tumorigenesis in vivo, the MSCs indirect co-cultured with C6 cells for 7?days were transplanted subcutaneously into immuno-deficient mice. The cells developed into a large tumour at the injection site within 8?weeks, with systemic symptoms including cachexia and scoliosis. Pathological and cytological analysis revealed poorly differentiated pleomorphic cells with a dense vascular network and aggressive invasion into the adjacent muscle. These data demonstrate that MSCs became malignant cancer cells when exposed to the tumour microenvironment and suggest that factors released from the cancer cells have a critical role in the malignant transformation of MSCs. Copyright (c) 2012 John Wiley & Sons, Ltd.