Thrombospondin promoted anti-tumor of adenovirus-mediated calreticulin in breast cancer: Relationship with anti-CD47
Thrombospondin promoted anti-tumor of adenovirus-mediated calreticulin in breast cancer: Relationship with anti-CD47
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DOI：
10.1016/j.biopha.2015.05.017
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发表时间：
2015-07-01
影响因子：
7.5
通讯作者：
Fang, Xudong
中科院分区：
文献类型：
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作者：
Chen, Qifeng;Fang, Xiaoming;Fang, Xudong
目的：钙网蛋白（Calreticulin， CRT）在肿瘤发生中具有多方面的作用，但其在乳腺癌中的作用尚不清楚。在这项研究中，我们试图评估过表达的CRT对乳腺癌细胞活力和增殖的影响。方法：分别采用定量RT-PCR法和Western blot法检测细胞中CRT和CD47 mRNA和蛋白的表达水平。流式细胞术Annexin V-FITC检测细胞凋亡。采用MTT法测定细胞活力。细胞迁移和自噬也被评估。结果：在MCF-7和MDA-MB-231乳腺癌细胞中，与MCF-10A正常乳腺细胞相比，CRT和CD47的表达均增强。转染MCF-7和MDA-MB-231的细胞过表达CRT，可显著抑制细胞迁移、抑制细胞活力，促进细胞凋亡，而对细胞自噬无影响。有趣的是，在MCF-7细胞异种移植中，血小板反应蛋白（TSP）和过表达CRT联合使用可显著诱导细胞自噬并抑制肿瘤生长。通过芯片实验，我们观察到TSP处理促进了TSP与CRT和CD47的相互作用。结论：TSP通过与CRT和CD47形成复合物促进腺病毒介导的CRT在乳腺癌中的抗肿瘤作用。(C) 2015 Elsevier Masson SAS。版权所有。
Objective: Calreticulin (CRT) protein has multifaceted role in carcinogenesis, however its role in breast cancer remains unidentified. In this study, we attempted to evaluate the effect of overexpressed CRT on breast cancer cells viability and proliferation.Methods: Levels of mRNA and protein expression for CRT and CD47 in cells were determined by Quantitative RT-PCR analysis and Western blot, respectively. Cells apoptosis was evaluated using Annexin V-FITC assay with flow cytometry. Cell viability was assessed using MTT assay. Cell migration and autophagy were also evaluated.Results: In breast cancer cells of MCF-7 and MDA-MB-231, both CRT and CD47 expression were enhanced, compared with that in normal breast cells of MCF-10A. Overexpression of CRT by MCF-7 and MDA-MB-231 cells transfected with significantly suppressed cell migration, viability as well as promote cell apoptosis while exerted no effected on cell autophagy. Interestingly, combining of thrombospondin (TSP) and overexpression of CRT significantly induced cell autophagy and inhibited tumor growth in MCF-7 cells xenograft. In result of chip assay, we observed that TSP treatment promoted interaction of TSP with CRT and CD47.Conclusion: TSP promoted anti-tumor of adenovirus-mediated CRT via forming complexes with CRT and CD47 in breast cancer. (C) 2015 Elsevier Masson SAS. All rights reserved.