基底刚度影响抗肿瘤药物效果的生物力药理学研究

Substrate stiffness as a crucially mechanical parameter within tumor microenvironments plays an important role in the pathophysiological process of tumor genesis and its development. This project intends to base on the theory framework of mechanopharmacology, focusing on the molecular mechanism on the antitumor drug response of tumor cells on different substrate stiffness by means of bioinformatics analysis methods. The plan includes the follow contents: (1) Fabricate the substrates with different stiffness by acrylamide gel and then culture the tumor cells on these stiffness-controlled substrates. (2) Carry out the identification of differentially expressed genes, bioinformatics analysis and validation in order to reveal the response of SK-BR-3 cells on the different substrate stiffness to antitumor drugs (including Cisplatin and Lapatinib). (3) Investigate the signaling pathways decided by specific genes which mediate the impact of substrate stiffness on tumor cell growth. (4) Clarify what are specific signaling pathways to mediate the response of SK-BR-3 cells cultured on different substrate stiffness to the antitumor drugs. Based on the results mentioned above, we can finally construct the signaling network of how the mechanical factors mediate both tumor genesis and its development, providing new targets for cancer early diagnosis, treatment and prognosis estimate. Meantime, this project may improve the better understanding of how the mechanical properties influent the response of tumor cells to antitumor drugs, developing a more rational and comprehensive antitumor drug screening system based on the substrates with variable stiffness, instead of the current conventional plastic culture plates in vitro.

基底刚度作为肿瘤微环境中重要的力学参数，在肿瘤发生、发展的病生理过程中起着重要的作用。本项目拟以生物力药理学理论为构架，运用生物信息学分析方法，着重研究基底刚度影响抗肿瘤药物效果的分子生物学机制。具体内容包括（1）制备可控刚度的丙烯酰胺凝胶基底，并培养肿瘤细胞；（2）进行不同刚度基底上抗肿瘤药物（顺铂、Lapatinib）作用与否SK-BR-3细胞内差异表达基因的鉴定、生物信息学的分析和验证；（3）探讨特定基因所在信号通路在刚度影响肿瘤细胞生长中的作用；（4）阐明特定基因所在信号通路在刚度影响抗肿瘤药物杀伤SK-BR-3细胞效果的作用。最终构建力学因素参与的肿瘤发生、发展特异性信号网络，为肿瘤早期诊断、治疗及预后改善提供新靶点。同时，确定特定基因及所在信号通路在力学因素影响肿瘤细胞性状及对抗肿瘤药物反应中的作用，拓展目前以塑料培养板为常规的体外药敏实验方法，建立更加合理和全面的药物筛选体系

基底刚度作为肿瘤微环境中重要的力学参数，在肿瘤发生、发展的病生理过程中起着重要的作用。本项目以生物力药理学理论为构架，应用基因表达谱芯片系统鉴定参与基底刚度影响乳腺癌SK-BR-3细胞生长及对靶向药物拉帕替尼（Lapatinib）反应的差异表达基因；并运用生物信息学分析方法表明差异表达基因主要富集的显著性功能和通路，探讨特定基因所在信号通路在刚度影响肿瘤细胞生长中的作用。最终构建力学因素参与的肿瘤发生、发展特异性信号网络，为肿瘤早期诊断、治疗及预后改善提供新靶点。同时，确定特定基因及所在信号通路在力学因素影响肿瘤细胞性状及对化疗药物反应中的作用，拓展目前以塑料培养板为常规的体外药敏实验方法，建立更加合理、更全面的药物筛选体系。.本文的主要结论是：.1. 基底刚度调节乳腺癌细胞的生长。.不同的基因参与基底刚度对乳腺癌细胞生长的过程，生物信息学分析差异表达基因体现的显著性功能和通路，PI-3K-AKT 和RAS-ERK信号通路可能参与基底刚度促进乳腺癌细胞生长过程。.2. 软基底降低乳腺癌细胞对Lapatinib 的反应.不同的基因参与基底刚度对乳腺癌细胞对Lapatinib 的反应的过程，生物信息学分析差异表达基因体现的显著性功能和通路，HER2信号通路可能参与这一反应过程。.另外，还增加了基于肿瘤周围筋膜力学环境改变的生物力药理学研究以及基底刚度调节肿瘤相关成纤维细胞影响肿瘤对化疗药物的反应。