低氧条件下RUNX2对小鼠乳腺癌4T1细胞凋亡的影响

发布时间：2018-06-02 11:05

本文选题：乳腺癌细胞 + Runt相关转录因子　；参考：《中国肿瘤生物治疗杂志》2017年04期

【摘要】：目的:探讨Runt相关转录因子2(Runt-related transcription factor 2,RUNX2)在低氧环境下对小鼠乳腺癌4T1细胞凋亡的影响及其作用机制。方法:Real-time PCR和Western blotting分别检测低氧条件对4T1细胞内RUNX1、RUNX2、RUNX3 mRNA和RUNX2蛋白表达的影响;采用小干扰RNA技术与真核重组质粒DNA过表达技术降低或者提高4T1细胞RUNX2的表达,免疫共沉淀法检测4T1细胞中RUNX2与其他蛋白之间的相互结合情况,流式细胞术检测常氧/低氧条件下干扰/过表达RUNX2对4T1细胞凋亡的影响。结果:低氧条件下4T1细胞中RUNX1和RUNX2 mRNA的表达水平上调(P0.05),RUNX2的蛋白表达量明显增加;转染RUNX2-siRNA-1415和真核表达载体pc DNA3.1(-)-RUNX2可使4T1细胞内RUNX2水平明显降低或升高;在常氧或低氧条件下,沉默RUNX2 mRNA的表达均导致小鼠乳腺癌4T1细胞凋亡率上升[常氧:(12.83±0.24)%vs(9.3±0.55)%,P0.05;低氧:(19.77±0.59)%vs(15.13±0.32)%,P0.05],而过表达RUNX2均导致4T1细胞凋亡率下降[常氧:(9.97±0.27)%vs(14.07±0.80)%,P0.05;低氧:(22.43±1.02)%vs(34.93±0.71)%,P0.05]。在低氧条件下,缺氧诱导因子-1α(hypoxia induced factor-1α,HIF-1α)与RUNX2的表达上升,RUNX2能与HIF-1α形成复合物。结论:在肿瘤低氧微环境中,RUNX2高表达于小鼠乳腺癌4T1细胞中,高表达的RUNX2可能是通过与HIF-1α的相互作用而抑制肿瘤细胞的凋亡。
[Abstract]:Aim: to investigate the effect and mechanism of Runt related transcription factor 2(Runt-related transcription factor 2 RUNX2 on apoptosis of mouse breast cancer 4T1 cells in hypoxic environment. Methods the effects of hypoxia on the expression of RUNX1, RUNX2, RUNX3 mRNA and RUNX2 protein in 4T1 cells were detected by 10: Real-time PCR and Western blotting, respectively, and the expression of RUNX2 in 4T1 cells was decreased or increased by using small interfering RNA technique and eukaryotic recombinant plasmid DNA overexpression technique. The interaction between RUNX2 and other proteins in 4T1 cells was detected by immunoprecipitation and the effect of interference / overexpression of RUNX2 on apoptosis of 4T1 cells under normoxic / hypoxic conditions was detected by flow cytometry. Results: the expression of RUNX1 and RUNX2 mRNA in 4T1 cells increased significantly under hypoxia, and the expression of RUNX2 in 4T1 cells was significantly decreased or increased after transfection of RUNX2-siRNA-1415 and eukaryotic expression vector PC DNA3.1(-)-RUNX2. Silencing the expression of RUNX2 mRNA increased the apoptosis rate of 4T1 cells in mouse breast cancer [normoxic RUNX2 mRNA 12.83 卤0.24)%vs(9.3 卤0.55; hypoxia: 19.77 卤0.59)%vs(15.13 卤0.32], while overexpression of RUNX2 resulted in a decrease of 4T1 cell apoptosis rate [normoxic 0.24)%vs(9.3 9.97 卤0.27)%vs(14.07 卤0.80 P0.05; hypoxia: 22.43 卤1.02)%vs(34.93 卤0.71]. Under hypoxia condition, the expression of hypoxia inducible factor 1 伪 hypoxia induced factor-1 伪 (HIF-1 伪) and RUNX2 increased. RUNX2 could form complex with HIF-1 伪. Conclusion: RUNX2 is highly expressed in mouse breast cancer 4T1 cells in hypoxic tumor microenvironment, and the overexpression of RUNX2 may inhibit the apoptosis of tumor cells through the interaction with HIF-1 伪.
【作者单位】：第二军医大学免疫学研究所暨医学免疫学国家重点实验室;
【基金】：国家高技术研究发展计划(863计划)课题资助项目(No.SS2014AA020801)~~
【分类号】：R392.12

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