基质力学微环境在上皮性卵巢癌耐药中的作用及机制研究

Tumor progression are followed by a range of physical changes in the tumor microenvironment such as increased matrix stiffness. As a crucial mechanical factor, stiffness of extracellular matrix with micro/nanoscaled structures plays an important role in many biological processes such as cell growth, proliferation, and differentiation. Recent studies by our research group have further indicated that mechanical changes in the extracellular matrix during tumor progression regulate the response to chemotherapeutics. we focus on epithelial ovarian cancer (EOC) and its drug resistance in clinical therapy. In the view of Biomechanopharmacology, we carry out the multiple levels of experiments, such as clinical sample, cell line and tumor inoculated animals. In the present study, we explore the effect of substrate stiffness on the drug resistance in EOC therapy, screening specific genes and responding signal pathways by means of the technologies from Biomechanopharmacology, clinical oncology, biomechanics, nano-high-resolution imaging, bioinformatics andmolecularbiology. Furthermore, we also develop an animal model for the study of mechanical microenvironment on the drug screening in vivo. Summarily, we try to develop a new strategy for correcting mechanical faults during cancer progression and adjusting signal pathways interacting with mechanical coupling to better promote the sensitivity of cancer therapy. In addition, it is promising for the development of more rational matrix for anti-tumour drug screening systems in vitro to replace the currently used plastic culture plates.

肿瘤力学微环境是目前肿瘤研究与诊疗的热点方向。本研究拟在前期基底刚度可影响肿瘤细胞化疗敏感性研究的基础上，针对上皮性卵巢癌化疗耐药问题，在生物力药理学理论指导下，从肿瘤微环境的力学因素--基底刚度入手，通过临床标本检测、体外细胞实验及接种移植瘤动物模型体内实验，运用生物力学、生物力药理学、纳米高分辨成像技术、生物信息学和分子生物学技术，研究基底刚度影响上皮性卵巢癌化疗敏感性的作用，筛选并进一步探讨特定基因及所在信号通路蛋白，寻找力学微环境影响化疗敏感性的新靶点，并建立实验动物模型，在体观察组织力学环境改变对药物治疗的影响，为早期预测卵巢癌的化疗耐药，筛选逆转耐药的新化疗方案，从而进行个体化治疗，改善卵巢癌预后奠定基础。

肿瘤微环境中的基底刚度在肿瘤发生、发展的病生理过程中起着重要的作用，还影响肿瘤细胞的化疗敏感性。为了探究基底刚度对卵巢癌发展和化疗耐药的影响，本项目制备了硬度不同的聚丙烯酰胺水凝胶模拟不同组织硬度，研究了不同基底硬度对卵巢癌细胞SKOV3的增殖和化疗药物顺铂和紫杉醇药效的影响，并利用多种生物技术和表征方法探究影响机制，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。.本课题的主要研究结论：.1.基底刚度调节卵巢癌细胞的生长..卵巢癌细胞呈现基底硬度依赖性生长。基底硬度增加，促进卵巢癌细胞生长，细胞在硬基底上增殖速率加快。通过研究不同硬度基底上YAP核定位情况发现硬基底上YAP在细胞核定位增加，表明硬基底上细胞生长加快可能是由于基底硬度增加促进YAP向细胞核易位，细胞转录活性增强，细胞增殖加快。.2.软基底降低卵巢癌对化疗药物的反应..软基底降低卵巢癌细胞对化疗药物的反应。通过研究不同硬度基底上卵巢癌细胞EMT影响发现，软基底上激活上皮间质转化的程序，细胞发生由上皮细胞向间质细胞转变。另外软基底上多药耐药蛋白表达增多。细胞硬度实验证实细胞的状态与硬基底相比，在软基底上表现为更软。.3.生物信息学分析基底硬度对细胞生长和化疗耐药差异表达基因的影响..不同的基因参与基底刚度卵巢细胞癌生长的过程，生物信息学分析差异表达基因体现的显著性功能和通路，细胞周期、MAPK、PI3K-Akt、Hippo信号通路可能参与基底刚度促进卵巢癌细胞生长过程。.不同的基因参与基底刚度对卵巢癌细胞对化疗药物紫杉醇及顺铂的反应的过程，抗肿瘤药耐药基因中的铂耐药基因、ABC运载体基因以及EGFR酪氨酸激酶抑制剂耐药基因可能参与调节基底刚度卵巢癌细胞对化疗药物的反应。