肿瘤细胞获得抗失巢凋亡能力是其发生转移的先决条件之一，阐明肿瘤细胞抗失巢凋亡分子机制有重要意义。我们前期研究结果发现小窝蛋白-1(cav-1)与乳腺癌细胞的生存和迁移能力密切相关，且切应力作用可以上调cav-1表达，但切应力条件下cav-1在肿瘤细胞抗失巢凋亡力学信号转导中的具体功能与调控机制仍不清楚。该项目采用悬浮培养切应力加载技术联合免疫细胞化学、免疫印迹及定量PCR等分析方法，研究切应力作用对肿瘤细胞膜上小窝形成及cav-1表达、分布、内化和活化的影响规律，重点探讨切应力如何调控cav-1功能改变及所引发的信号转导途径、整合素beta(1)亲和性、及抗细胞凋亡相关基因表达水平，揭示cav-1在肿瘤细胞抗失巢凋亡力学信号转导中的功能与影响机制，阐明切应力和cav-1与细胞抗失巢凋亡的内在关系和贡献大小，从而开辟遏制肿瘤细胞抗失巢凋亡能力的新途径，为防治肿瘤转移和抗癌药物开发提供新靶点。

Hematogenous metastasis is the mainly leading cause of death in cancer patients, which is a multistep process including dissociation of cancer cells from primary sites, survival in the vascular system, and proliferation in distant target organs. The acquisition of anoikis resistance (anti-anoikis) for metastatic cancer cells is required for hematogenous metastasis. Cancer cells acquire anoikis resistance to survive after detachment from the primary sites and travel through the circulatory and lymphatic systems to disseminate throughout the body. Currently, anoikis resistance becomes a hot topic in cancer research. A better understanding of the underlying molecular and cellular mechanisms is crucial for the development of effective treatment for metastatic cancer. Our previous studies demonstrated that caveolin-1 (cav-1) is related to cell survive and migration of human breast cancer cells (MDA-MB-231), and low shear stress up-regulated cav-1 expression. However, the mechanobiological roles of cav-1 in anti-anoikis of human breast cancer cells under shear flow conditions and its mechanotransduction or regulatory mechanisms are not well understood and remain to be elucidated. In this project, we will firstly investigate the effects of low shear stress on the expression, distribution, internalization and activation of cav-1 by suspenssion cultured shear stress loading apparatus (Couette rotation system) combining immunofluorescence cytochemistry, real time fluorescence quantitative PCR and western blot. Then we further explore how the low shear stress regulates cav-1 function and its triggered mechanotransduction pathways. The affinity of beta(1) integrin, FAK/beta(1) integrin interaction, and the expression of anti-apoptosis related genes will be also investigated after MDA-MB-231 cells exposure to low shear stress. Through the above-mentioned experiments, we want to elucidate the relationship between of cav-1 and anti-anoikis, and the mechanobiological roles and regulatory mechanisms of cav-1 in anti-anoikis of cancer cells under shear flow conditions. The detailed contribution of cav-1 and shear stress to anti-anoikis of human breast cancer cells, and their internal relationships will be illuminated through this proposed project investigation. These studies will provide insight into the biology of cancer metastasis and identify novel therapeutic targets for prevention of cancer dissemination.