中国肿瘤生物治疗杂志2017，Vol.24Issue(4)：373-379,7.DOI:10.3872/j.issn.1007-385X.2017.04.007
低氧条件下RUNX2对小鼠乳腺癌4T1细胞凋亡的影响
The effect of RUNX2 on apoptosis of rat breast cancer 4T1 cells under hypoxia
摘要
Abstract
Objective:To explore the effect of Runt-related transcription factor 2 (RUNX2) on apoptosis of mouse breast cancer 4T1 cells under hypoxia and its mechanism.Methods:Western blotting and Real-time PCR were used to measure the effect of hypoxia on the expression levels of RUNX1,RUNX2,RUNX3 mRNA and RUNX2 protein in 4T1 cells;small interference RNA (siRNA) and eukaryotic recombinant plasmid DNA over-expression technique were used to down-or up-regulate the RUNX2 expression in 4T1 cells respectively;Co-immunoprecipitation was used to detect the binding of RUNX2 and other proteins in 4T1 cells;Flow cytometry was used for the detection of the effect of down-/up-regulation of RUNX2 on apoptosis of 4T1 cells under normoxic/hypoxic condition.Results:The mRNA expression levels of RUNX1 and RUNX2 in 4T1 cells were up-regulated under hypoxia condition (P ＜ 0.05),and the protein expression of RUNX2 was significantly elevated (P ＜ 0.05).Transfection with RUNX2-siRNA-1415 or pcDNA3.1 (-)-RUNX2 could significantly decrease or increase the RUNX2 level in 4T1 cells,respectively;Under normoxic/hypoxic condition,apoptosis rate of 4T1 cells that transfected with siRNA-RUNX2-1415 was significantly increased by comparing with negative control group(normoxia:[12.83 ±0.2404]％ vs [9.3 ±0.5508]％,P＜0.05;hypoxia:[19.77 ±0.59]％ vs [15.13 ± 0.32] ％,P ＜ 0.05);however,the apoptosis rate of 4T1 cells were decreased by over-expressing RUNX2 (normoxia:[9.967 ± 0.2728] ％ vs [14.07 ± 0.7965]％,P＜0.05;hypoxia:[22.43 ±1.02]％ vs [34.93 ±0.71]％,P＜ 0.05).Under hypoxic condition,the expression levels of hypoxic induced factor-1α (HIF-1α) and RUNX2 were significantly elevated,and RUNX2 could bind with HIF-1α to form complex.Conclusion:Under the hypoxia micro-environment,4T1 cells express the high level of RUNX2,in the meanwhile,high level RUNX2 could inhibit the apoptosis of tumor cells by interacting with HIF-1α.关键词
乳腺癌细胞/Runt相关转录因子2/RNA干扰/低氧环境/细胞凋亡Key words
breast cancer cells/Runt-related transcription factor 2 (RUNX2)/RNA interference/Hypoxia environment/cell apoptosis分类
医药卫生引用本文复制引用
林妘,徐双,钱程..低氧条件下RUNX2对小鼠乳腺癌4T1细胞凋亡的影响[J].中国肿瘤生物治疗杂志,2017,24(4):373-379,7.基金项目
国家高技术研究发展计划(863计划)课题资助项目(No.SS2014AA020801).Project supported by the National High Technology Research and Development Program of China (863 Program) (No.SS2014AA020801) （863计划）
