Chen Pingbo, Mo Qingqing, Wang Beibei, Weng Danhui, Wu Peng, Chen Gang
Department of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
J Huazhong Univ Sci Technolog Med Sci. 2012 Feb;32(1):92-96. doi: 10.1007/s11596-012-0016-8. Epub 2012 Jan 27.
This study was aimed to explore the influence of breast cancer associated fibroblasts (CAFs) in migration and invasion of breast cancer cell line MCF-7, and investigate whether hepatocyte growth factor (HGF) is involved in this process. Primary breast CAFs and their corresponding normal breast fibroblasts (NFs) were obtained by collagenase digestion. On the basis of the co-culture, the migration and invasion capacity of MCF-7 cells was compared between CAFs and NFs by Transwell. The difference in the HGF expression between them was detected by ELISA. The secretion of HGF was knocked down by using RNA interference technology in CAFs. Then the changes of migration and invasion capacity of MCF-7 cells were investigated by Transwell. Eventually, we isolated high-purity CAFs and NFs, and the CAFs had a stronger ability in promoting MCF-7 migration and invasion than the NFs. ELISA results demonstrated that CAFs secreted higher HGF, and the capacity of MCF-7 migration and invasion was declined after knocking down the secretion of HGF in CAFs by RNA interference. It is suggested that CAFs can promote MCF-7 migration and invasion through HGF in vitro.
本研究旨在探讨乳腺癌相关成纤维细胞(CAFs)对乳腺癌细胞系MCF-7迁移和侵袭的影响,并研究肝细胞生长因子(HGF)是否参与这一过程。通过胶原酶消化获取原代乳腺CAFs及其相应的正常乳腺成纤维细胞(NFs)。在共培养的基础上,采用Transwell法比较CAFs和NFs对MCF-7细胞迁移和侵袭能力的影响。通过ELISA检测它们之间HGF表达的差异。利用RNA干扰技术敲低CAFs中HGF的分泌。然后采用Transwell法研究MCF-7细胞迁移和侵袭能力的变化。最终,我们分离出了高纯度的CAFs和NFs,且CAFs促进MCF-7迁移和侵袭的能力强于NFs。ELISA结果表明,CAFs分泌的HGF更高,通过RNA干扰敲低CAFs中HGF的分泌后,MCF-7的迁移和侵袭能力下降。提示在体外CAFs可通过HGF促进MCF-7的迁移和侵袭。