摘要
该文探讨了8-氯腺苷(8-chloro-adenosine, 8-Cl-Ado)调控RNA编辑酶1(adenosine deaminases acting on RNA1, ADAR1)对乳腺癌细胞增殖、迁移和侵袭的影响,确定了ADAR1与miR-335-5p表达的相关性。10μmol/L的8-Cl-Ado作用于乳腺癌细胞后(不同时间点),采用CCK-8检测细胞的增殖情况;Transwell检测细胞的迁移和侵袭情况;Western blot检测ADAR1蛋白的表达水平;CCK-8检测ADAR1过表达对8-Cl-Ado抑制乳腺癌细胞增殖的影响;Transwell检测ADAR1过表达对8-Cl-Ado抑制乳腺癌细胞迁移和侵袭的影响;miRNA芯片筛选8-Cl-Ado处理后乳腺癌细胞中上调的miRNA, qRT-PCR验证其与ADAR1的相关性。结果显示, 8-Cl-Ado能抑制乳腺癌细胞的增殖、迁移和侵袭;ADAR1蛋白的表达量随8-Cl-Ado处理时间的增加而逐渐降低;ADAR1的过表达能减弱8-Cl-Ado对乳腺癌细胞增殖、迁移和侵袭的抑制作用;miR-335-5p经8-Cl-Ado处理后表达水平上调,与ADAR1呈负向调控关系。以上结果表明, 8-Cl-Ado下调ADAR1抑制乳腺癌细胞增殖、迁移和侵袭,其作用机制可能与ADAR1下调mi R-335-5p有关。
The aim of this study was to investigate the effects of 8-chloro-adenosine(8-Cl-Ado)on proliferation,migration and invasion of breast cancer cells by regulating adenosine deaminases acting on RNA1(ADAR1),and the correlation of expression between ADAR1 and miR-335-5 p.After the treatment of breast cancer cells with10μmol/L of 8-Cl-Ado at different time points or overexpression of ADAR1 in the cells for 48 h,proliferation was examined by CCK-8 while migration and invasion were determined by Transwell,and the expression levels of ADAR1 were measured by Western blot.Up-regulated miRNAs were screened by miRNA microarray and confirmed by qRT-PCR.The results showed that 8-Cl-Ado inhibited the proliferation,migration and invasion of breast cancer cells.The protein expression of ADAR1 gradually decreased in a time dependent manner.ADAR1 overexpression reduced the inhibition of proliferation,migration and invasion caused by 8-Cl-Ado.MiR-335-5 p was highly expressed after 8-Cl-Ado treatment,and it was negatively regulated by ADAR1.In summary,8-Cl-Ado inhibits proliferation,migration and invasion of breast cancer cells through down-regulating ADAR1,and its mechanism may relate to inhibition of miR-335-5 p by ADAR1.
作者
张梨虹
涂增
谢凤
杨生永
李梨
Zhang Lihong;Tu Zeng;Xie Feng;Yang Shengyong;Li Li(Molecular Medicine and Cancer Research Centre,Chongqing Medical University,Chongqing 400016,China)
出处
《中国细胞生物学学报》
CAS
CSCD
2019年第5期875-882,共8页
Chinese Journal of Cell Biology
基金
国家自然科学基金(批准号:81872444)
重庆市渝中区科技计划项目(批准号:20170109)资助的课题~~
