New mouse xenograft model modulated by tumor-associated fibroblasts for human multi-drug resistance in cancer

Ma,Yan; Lin,Zhiqiang; Fallon,John  K.; Zhao,Qiang; Liu,Dan; Wang,Yongjun; Liu,Feng

doi:10.3892/or.2015.4265

New mouse xenograft model modulated by tumor-associated fibroblasts for human multi-drug resistance in cancer

Authors:
- Yan Ma
- Zhiqiang Lin
- John K. Fallon
- Qiang Zhao
- Dan Liu
- Yongjun Wang
- Feng Liu
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Affiliations: School of Chinese Materia Medica, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510006, P.R. China, Department of Pharmaceutics, School of Pharmaceutical Sciences, Peking University, Beijing 100191, P.R. China, Division of Molecular Pharmaceutics, Eshelman School of Pharmacy, University of North Carolina, Chapel Hill, NC, USA, College of Chemical Engineering, Sichuan University, Chengdu, Sichuan 610065, P.R. China, Key Laboratory of Structure‑Based Drug Design and Discovery (Shenyang Pharmaceutical University), Ministry of Education, Shenyang, Liaoning 110016, P.R. China, School of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, P.R. China
Published online on: September 9, 2015 https://doi.org/10.3892/or.2015.4265
Pages: 2699-2705

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Abstract

We developed an MDR tumor model that is modulated by tumor-associated fibroblasts. Studies on proliferation of tumor cell lines including paclitaxel-sensitive and resistant cell lines were performed. The expressions of P-gp and α-smooth muscle actin (α-SMA) antigen were evaluated by immunohistochemistry and western blot analysis. Quantitative P-gp analyses of different cell lines were accomplished by nanoUPLC‑MS/MS. Tumor cell colony formation assay and established xenograft model was used to investigate the relationship between P-gp expression, fibroblast levels and tumorigenesis. The mouse xenograft model was developed after co-inoculation with MDR tumor cells and NIH/3T3 fibroblast cells. There was no correlation between tumorigenesis in vivo and the growth rate of cells in vitro. The proliferation among different cell lines had no significant differences, but the P-gp expression and tumor growth in the xenograft model were fairly different. P-gp determination and α-SMA immunofluorescence staining clarified the relationship between P-gp expression, fibroblast levels and tumorigenesis. It was more difficult for tumor cells with higher P-gp levels to recruit fibroblasts in vivo, resulting in lower tumorigenesis due to the lack of structural and chemical support during tumor progression. In the established paclitaxel-resistant mouse xenograft model, no obvious antitumor effect was observed after Taxol treatment, but a significant decrease in tumor size for the group treated with gemcitabine sensitive to the model. The results show that the added fibroblasts do not disturb the applicability of the model in MDR. Therefore, this mouse xenograft MDR model could serve as an effective tool for MDR research.

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