摘要
该文旨在探索敲低整合素相关蛋白CD47对卵巢癌细胞生物学行为和功能的作用。在卵巢癌细胞转染靶向CD47的小干扰RNA(small interfering RNA,siRNA)后,采用Western blotting、qRT-PCR、流式细胞术验证敲低效率;采用CCK-8、划痕实验、Transwell观察细胞增殖、迁移和侵袭情况;采用qRT-PCR检测细胞中CD47、IL-6、C-X-C基序趋化因子配体1(C-X-C motif chemokine ligand 1,CXCL1)、CXCL8、C-C基序趋化因子配体20(C-C motif chemokine ligand 20,CCL20)mRNA表达;采用ELISA检测细胞上清IL-6和CXCL8的释放;采用Transwell观察敲低CD47后卵巢癌细胞对巨噬细胞迁移的影响。结果发现敲低CD47分子对卵巢癌细胞增殖、迁移和侵袭无明显作用;而敲低CD47可降低卵巢癌细胞趋化因子CXCL1、CXCL3、CXCL8、CCL20和IL-6的表达,且明显地降低巨噬细胞的迁移能力。该研究表明卵巢癌细胞中CD47分子能够通过调节自身趋化因子表达从而影响巨噬细胞的迁移能力。
To investigate the effect of CD47 knockdown on the migration,proliferation,and chemokine secretion of ovarian cancer cells,small interfering RNA(siRNA)targeting CD47 was transfected to ovarian cancer cells and the knockdown efficiency was confirmed by Western blotting,qRT-PCR,and flow cytometry.Cell proliferation,migration,and invasion were examined using CCK-8,wound healing assay,and the Transwell assay,respectively.qRT-PCR was used to detect the expressions of CD47,C-X-C motif chemokine ligand 1(CXCL1),CXCL8,C-C motif chemokine ligand 20(CCL20)and IL-6.ELISA was used to determine the release of IL-6 and CXCL8.Transwell was employed to study macrophage invasion of CD47 knockdown ovarian cancer cells.The results showed that the expressions of chemokines CXCL1,CXCL3,CXCL8,CCL20,and IL-6 in ovarian cancer cells were reduced when CD47 was knocked down,along with the reduction of the migration capacity of macrophages towards ovarian cancer cells.CD47 knockdown did not affect the proliferation,migration,or invasion of ovarian cancer cells.Our findings suggest that CD47 in ovarian cancer cells can affect the migration ability of macrophages by regulating their chemokine expression.
作者
韩仕阳
王小瑜
傅丰庆
周淑如
HAN Shi-yang;WANG Xiao-yu;FU Feng-qing;ZHOU Shu-ru(Aoyang Cancer Institute,Affiliated Aoyang Hospital of Jiangsu University,Zhangjiagang 215600,China;Jiangsu Institute of Clinical Immunology,the First Affiliated Hospital of Soochow University,Suzhou 215000,China)
出处
《现代免疫学》
CAS
2024年第1期10-16,共7页
Current Immunology
基金
张家港市科技项目(ZKS1930)
江苏大学临床医学科技发展基金(JLY2021100)
苏州市民生科技项目(SYSD2020011)。
