中国药理学通报2016，Vol.32Issue(3)：323-326,327,5.DOI:10.3969/j.issn.1001-1978.2016.03.006
钠钾ATP酶抑制剂通过调节DNA损伤感应复合体Mre11/Rad50/Nbs1的表达诱导肝癌HepG2细胞周期阻滞
Na+,K+-ATPase inhibitor induces cell cycle arrest in liver cancer HepG2 cells by regulating expression of DNA damage Mre11/Rad50/Nbs1 complex
摘要
Abstract
Aim To explore the relationship between Mre11/Rad50/Nbs1 ( MRN ) complex focus formation and DNA double-strand breaks( DSBs) caused by cinob-ufagin in human hepatocellular carcinoma HepG2 cells. Methods The Na+,K+-ATPaseα1 subunit expression level in liver cancer tissues was detected by immunohis-tochemistry. After HepG2 cells were treated with 5μmol·L-1 cinobufagin for 6, 12 and 24 h, the drug-in-duced DSBs were assessed by single cell gel electro-phroesis ( SCGE ) , the gene transcription and protein levels of Mrel1, Nbs1, Rad50 and p53 were evaluated by Real time-PCR and Western blot. The cell cycle in parallel was analyzed by flow cytometry. Results The Na+, K+-ATPase α1 subunit expression level in liver cancer tissues was significantly increased compared with the tissue adjacent to carcinoma ( P <0. 05 ) . The 5μmol · L-1 cinobufagin could induce the DSBs in a time-dependent manner (P <0. 05), and it could up-regulate the gene expression levels of Mre11, Nbs1, Rad50 and p53 in HepG2 cells ( P<0. 05 ) . The pro-portions of HepG2 cells in S phase were ( 21. 32 ± 4. 21) % in the control group, and (33. 25 ± 5. 72) %, (56. 72 ± 6. 29) % and (67. 32 ± 9. 42) % in HepG2 cells treated with 5 μmol · L-1 cinobufagin for 6, 12 and 24 h, respectively. The proportions of cells in S phase in cinobufagin groups were significantly increased compared with the control group ( P<0. 05 ) . Conclu-sion Cinobufagin could induce the cell cycle arrest in liver cancer HepG2 cells by activation of Mre11/Rad50/Nbs1 Complex.关键词
华蟾毒配基/HepG2细胞/DNA双链断裂/钠钾ATP酶/细胞周期/细胞周期相关蛋白Key words
cinobufagin/HepG2 cells/DNA double-strand breaks/Na+, K+-ATPase/cell cycle/cell cycle associated proteins分类
医药卫生引用本文复制引用
徐忠伟,王凤梅,王聪聪,单娜娜,徐瑞成..钠钾ATP酶抑制剂通过调节DNA损伤感应复合体Mre11/Rad50/Nbs1的表达诱导肝癌HepG2细胞周期阻滞[J].中国药理学通报,2016,32(3):323-326,327,5.基金项目
国家自然科学基金资助项目( No 81273552) （ No 81273552）
武警后勤学院博士启动金项目(No WHB201501) （No WHB201501）
