间充质干细胞对小鼠体内转移发展的影响。
Influence of mesenchymal stem cells on metastasis development in mice in vivo.
作者信息
Meleshina Aleksandra V, Cherkasova Elena I, Shirmanova Marina V, Klementieva Natalia V, Kiseleva Ekaterina V, Snopova Ludmila В, Prodanets Natalia N, Zagaynova Elena V
机构信息
Nizhny Novgorod State Medical Academy, Minin and Pozharsky Square, 10/1, Nizhny, Novgorod, 603005, Russia.
Nizhny Novgorod State University, Gagarin Avenue, 23, Nizhny, Novgorod, 603950, Russia.
出版信息
Stem Cell Res Ther. 2015 Feb 27;6(1):15. doi: 10.1186/s13287-015-0003-7.
INTRODUCTION
In recent years, mesenchymal stem cells (MSCs) have been demonstrated to play an important role in carcinogenesis. However, the effect of MSCs on tumor and metastasis development and the mechanisms underlying the interaction of cancer and stem cells are not completely understood. This study investigated the effect of MSCs on breast cancer metastasis formation by using the methods of in vivo fluorescence and luminescence imaging.
METHODS
MSCs were isolated from bone marrow of normal donors, characterized, and genetically labeled with luciferase (luc2). The effects of MSCs on MDA-MB-231 cancer cell proliferation were evaluated in conditioned medium from MSCs. To generate lung metastases, MDA-MB-231 cells stably expressing red fluorescent protein Turbo FP650 were injected intravenously into nude mice. On day 10 after the cancer cell injection, mice were injected via the tail vein with MSCs-luc2 cells (the MET+MSCs group). Animals that received the injection of MDA-MB-231-Turbo FP650 alone (the MET group) and no injections (the intact control group) served as controls. Fluorescence and bioluminescence imaging was performed for monitoring of the metastasis formation and MSC distribution in the recipient's body.
RESULTS
We found that the proliferative activity of the cancer cells in the presence of MSC conditioned medium was lower than that of the cells grown in conventional culture medium. The metastasis formation in the MET+MSCs group was delayed in time as compared with the MET group. Macroscopic and histological examination of isolated lungs 8 weeks after cancer cell injection showed that the total number of metastases in animals of the MET+MSCs group was significantly lower. Using bioluminescence imaging in vivo, we found that MSCs-luc2 cells survived in the host animal for at least 7 weeks and re-migrated to the lung 6 to 7 weeks after injection. Immunohistochemical analysis revealed the presence of MSCs-luc2 in metastases and lung tissue.
CONCLUSIONS
Long-term in vivo bioluminescence imaging of intravenously injected MSCs-luc2 cells showed distribution of MSCs to the lungs and abdominal organs within the first 2 to 3 weeks and re-migration to the lungs in weeks 6 to 7. It was found that MSCs reduced the proliferative activity of cancer cells in vitro and lung metastasis formation in mice.
引言
近年来，间充质干细胞（MSCs）已被证明在肿瘤发生过程中发挥重要作用。然而，MSCs对肿瘤及转移发展的影响以及癌症与干细胞相互作用的潜在机制尚未完全明确。本研究采用体内荧光和发光成像方法，研究了MSCs对乳腺癌转移形成的影响。
方法
从正常供体的骨髓中分离出MSCs，进行鉴定，并用荧光素酶（luc2）进行基因标记。在MSCs的条件培养基中评估MSCs对MDA-MB-231癌细胞增殖的影响。为了产生肺转移，将稳定表达红色荧光蛋白Turbo FP650的MDA-MB-231细胞静脉注射到裸鼠体内。在癌细胞注射后第10天，通过尾静脉给小鼠注射MSCs-luc2细胞（MET+MSCs组）。仅接受MDA-MB-231-Turbo FP650注射的动物（MET组）和未注射的动物（完整对照组）作为对照。进行荧光和生物发光成像，以监测受体体内转移灶的形成和MSCs的分布。
结果
我们发现，在存在MSCs条件培养基的情况下，癌细胞的增殖活性低于在传统培养基中生长的细胞。与MET组相比，MET+MSCs组的转移形成时间延迟。在癌细胞注射8周后对分离的肺进行大体和组织学检查发现，MET+MSCs组动物的转移灶总数明显减少。通过体内生物发光成像，我们发现MSCs-luc2细胞在宿主动物体内存活至少7周，并在注射后6至7周重新迁移到肺部。免疫组织化学分析显示转移灶和肺组织中存在MSCs-luc2。
结论
静脉注射MSCs-luc2细胞的长期体内生物发光成像显示，MSCs在最初2至3周内分布到肺部和腹部器官，并在第6至7周重新迁移到肺部。研究发现，MSCs在体外降低了癌细胞的增殖活性，并减少了小鼠的肺转移形成。
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