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间质干细胞向肿瘤异种移植模型的迁移和阿霉素的药物传递。

Migration of mesenchymal stem cells to tumor xenograft models and drug delivery by doxorubicin.

机构信息

Department of Nuclear Medicine, School of Medicine, Kyungpook National University, Kyungpook National University Hospital, Daegu, Republic of Korea.

出版信息

Int J Med Sci. 2018 Jun 22;15(10):1051-1061. doi: 10.7150/ijms.25760. eCollection 2018.

DOI:10.7150/ijms.25760
PMID:30013447
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6036160/
Abstract

Mesenchymal stem cells (MSCs) show therapeutic effects in various types of diseases. MSCs have been shown to migrate towards inflamed or cancerous tissues, and visualized after sacrificing the animal. MSCs are able to deliver drugs to target cells, and are an ideal candidate for cancer therapy. The purpose of this study was to track the migration of MSCs in tumor-bearing mice; MSCs were also used as drug delivery vehicles. Human breast cancer cells (MDA-MB-231) and anaplastic thyroid cancer cells (CAL62) were transduced with lentiviral particles, to express the luciferase and mCherry (mCherry-Rluc) reporter genes. Human bone marrow-derived MSCs were transduced with lentiviral particles, to express the firefly luciferase and enhanced green fluorescence protein (Fluc2-eGFP) reporter genes (MSC/Fluc). Luciferase activity of the transduced cells was measured by bioluminescence imaging (BLI). Further migration assays were performed to confirm cancer cells conditioned medium dependent MSC and doxorubicin (DOX) treated MSC migration. MSCs were loaded with DOX, and their therapeutic effects against the cancer cells were studied . MSC/Fluc migration in mice having thyroid or breast cancer xenografts was evaluated after systemic injection. Rluc activity of CAL62/Rluc (R=0.911), MDA-MB-231/Rluc (R=0.934) cells and Fluc activity of MSC/Fluc (R=0.91) cells increased with increasing cell numbers, as seen by BLI. eGFP expression of MSC/Fluc was confirmed by confocal microscopy. Similar migration potential was observed between MSC/Fluc and naïve MSCs in migration assay. DOX treated MSCs migration was not decreased compared than MSCs. Migration of the systemically injected MSC/Fluc cells into tumor xenografts (thyroid and breast cancer) was visualized in animal models (<0.05) and confirmed by (<0.05) BLI. Additionally, MSCs delivered DOX to CAL62/Rluc and MDA-MB-231/Rluc cells, thereby decreasing their Rluc activities. In this study, we confirmed the migration of MSCs to tumor sites in cancer xenograft models using both and BLI imaging. DOX-pretreated MSCs showed enhanced cytotoxic effects. Therefore, this noninvasive reporter gene (Fluc2)-based BLI may be useful for visualizing tracking of MSCs, which can be used as a drug delivery vehicle for cancer therapy.

摘要

间充质干细胞（MSCs）在各种类型的疾病中显示出治疗效果。已经表明 MSCs 向炎症或癌组织迁移，并在牺牲动物后可以可视化。MSCs 能够将药物递送到靶细胞，是癌症治疗的理想候选物。本研究的目的是跟踪肿瘤荷瘤小鼠中 MSCs 的迁移；MSCs 也被用作药物输送载体。人乳腺癌细胞（MDA-MB-231）和间变性甲状腺癌细胞（CAL62）被慢病毒颗粒转导，以表达荧光素酶和 mCherry（mCherry-Rluc）报告基因。人骨髓来源的 MSCs 被慢病毒颗粒转导，以表达萤火虫荧光素酶和增强型绿色荧光蛋白（Fluc2-eGFP）报告基因（MSC/Fluc）。通过生物发光成像（BLI）测量转导细胞的荧光素酶活性。进一步进行迁移实验以确认癌细胞条件培养基依赖性 MSC 和多柔比星（DOX）处理 MSC 的迁移。将 DOX 加载到 MSC 中，并研究其对癌细胞的治疗效果。在系统注射后评估具有甲状腺或乳腺癌异种移植物的小鼠中 MSC/Fluc 的迁移。通过 BLI 观察到，CAL62/Rluc（R=0.911）、MDA-MB-231/Rluc（R=0.934）细胞的 Rluc 活性和 MSC/Fluc 的 Fluc 活性随细胞数量的增加而增加。通过共聚焦显微镜证实了 MSC/Fluc 的 eGFP 表达。在迁移实验中观察到 MSC/Fluc 与未处理的 MSC 之间具有相似的迁移潜力。与 MSC 相比，DOX 处理的 MSC 的迁移没有减少。在动物模型中观察到系统注射的 MSC/Fluc 细胞进入肿瘤异种移植物（甲状腺和乳腺癌）的迁移（<0.05），并通过（<0.05）BLI 证实。此外，MSCs 将 DOX 递送至 CAL62/Rluc 和 MDA-MB-231/Rluc 细胞，从而降低了它们的 Rluc 活性。在这项研究中，我们使用和 BLI 成像确认了 MSCs 在癌症异种移植模型中向肿瘤部位的迁移。DOX 预处理的 MSC 显示出增强的细胞毒性作用。因此，这种非侵入性报告基因（Fluc2）-基于 BLI 可能可用于可视化和跟踪 MSC，可将其用作癌症治疗的药物输送载体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7cde/6036160/2350e1ebc6c9/ijmsv15p1051g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7cde/6036160/e642abe624db/ijmsv15p1051g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7cde/6036160/b28c9ec75861/ijmsv15p1051g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7cde/6036160/9c1e793c775a/ijmsv15p1051g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7cde/6036160/59123d59ab0e/ijmsv15p1051g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7cde/6036160/2350e1ebc6c9/ijmsv15p1051g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7cde/6036160/e642abe624db/ijmsv15p1051g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7cde/6036160/b28c9ec75861/ijmsv15p1051g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7cde/6036160/9c1e793c775a/ijmsv15p1051g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7cde/6036160/59123d59ab0e/ijmsv15p1051g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7cde/6036160/2350e1ebc6c9/ijmsv15p1051g006.jpg

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本文引用的文献

[1]
Drug Discovery by Molecular Imaging and Monitoring Therapy Response in Lymphoma.

Int J Mol Sci. 2017-7-27

[2]
In Vivo Tracking of Chemokine Receptor CXCR4-Engineered Mesenchymal Stem Cell Migration by Optical Molecular Imaging.

Stem Cells Int. 2017

[3]
Genetically engineered suicide gene in mesenchymal stem cells using a Tet-On system for anaplastic thyroid cancer.

PLoS One. 2017-7-20

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