衰老上皮细胞及衰老成纤维细胞的分子通话对邻近上皮细胞侵袭迁移的影响及其关键分子初探

项目名称： 衰老上皮细胞及衰老成纤维细胞的分子通话对邻近上皮细胞侵袭迁移的影响及其关键分子初探

项目编号： No.31301141

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 生物科学

项目作者： 刘洁薇

作者单位： 四川大学

项目金额： 20万元

中文摘要： 肿瘤及其微环境衰老对肿瘤的发生发展均发挥着重要作用。本人的前期研究表明衰老成纤维细胞可通过分泌细胞因子促进邻近上皮细胞的侵袭迁移。然而，多种细胞的共同衰老是否具有协同/相加/拮抗作用，国内外尚无相关报道。本研究拟运用博来霉素诱导人上皮细胞HMEC及其相关成纤维细胞株RMF衰老，运用Transwell迁移小室检测衰老HMEC/衰老RMF/共培养双衰老细胞的条件培养液对未衰老HMEC的促迁移作用；运用三维细胞共培养技术观察上述各组衰老细胞对未衰老HMEC的促侵袭作用；将衰老前后的两种细胞两两组合进行二维共培养，运用生长因子抗体芯片检测各组条件培养液并筛选出候选关键因子。运用ELISA法及实时荧光RT-PCR法确认，并在上述迁移小室及三维共培养体系中运用相关抑制剂验证。以明确多种衰老细胞的相互作用对细胞群体侵袭迁移的影响，为细胞衰老在肿瘤微环境中的研究提供新模型新思路。

中文关键词： 细胞衰老；肿瘤微环境；细胞增殖；细胞迁移；

英文摘要： The senescence in tumor and associated microenvironment plays an important role in tumor initiation, progression, invasion and metastasis. Our previous research has revealed the secretary cytokines from senescent fibroblasts which promoted migration and invasion of associated epithelial cells. However, whether there is an synergism，addition，or antagonism effect among different cell types that have gone through senescence has not been investigated at home and abroad. This research is therefore designed to induce senescence in human mammary epithelial cells (HMEC) and human reduced mammary fibroblasts (RMF) by bleomycin. Migration assay by traswell chambers will be performed on presenescent HMECs using condition medium of senescent HMEC or RMF or coculture of senescent HMEC and RMF cells. Matrigel 3D coculture will be performed on presenescent HMECs with either of the three senescent groups metioned above. The pre-/post-senescent HMECs and RMFs will be cocultured pairwisely in a 2D manner in order to collect the condition medium. Cytokine antibody array will be performed and candidate cytokine that is most likely to be associated will be chosen. Enzyme-linked immuno sorbent assay (ELISA) and real time RT-PCR will be performed to validate the result. Transwell migration assay and matrigel 3D coculture will be re

英文关键词： Tumor senescence；Tumor microenvironment；proliferation；migration；