本文選題：乳腺癌細胞 + Runt相關轉(zhuǎn)錄因子��； 參考：《中國腫瘤生物治療雜志》2017年04期

【摘要】：目的:探討Runt相關轉(zhuǎn)錄因子2(Runt-related transcription factor 2,RUNX2)在低氧環(huán)境下對小鼠乳腺癌4T1細胞凋亡的影響及其作用機制。方法:Real-time PCR和Western blotting分別檢測低氧條件對4T1細胞內(nèi)RUNX1、RUNX2、RUNX3 mRNA和RUNX2蛋白表達的影響;采用小干擾RNA技術與真核重組質(zhì)粒DNA過表達技術降低或者提高4T1細胞RUNX2的表達,免疫共沉淀法檢測4T1細胞中RUNX2與其他蛋白之間的相互結(jié)合情況,流式細胞術檢測常氧/低氧條件下干擾/過表達RUNX2對4T1細胞凋亡的影響。結(jié)果:低氧條件下4T1細胞中RUNX1和RUNX2 mRNA的表達水平上調(diào)(P0.05),RUNX2的蛋白表達量明顯增加;轉(zhuǎn)染RUNX2-siRNA-1415和真核表達載體pc DNA3.1(-)-RUNX2可使4T1細胞內(nèi)RUNX2水平明顯降低或升高;在常氧或低氧條件下,沉默RUNX2 mRNA的表達均導致小鼠乳腺癌4T1細胞凋亡率上升[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],而過表達RUNX2均導致4T1細胞凋亡率下降[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]。在低氧條件下,缺氧誘導因子-1α(hypoxia induced factor-1α,HIF-1α)與RUNX2的表達上升,RUNX2能與HIF-1α形成復合物。結(jié)論:在腫瘤低氧微環(huán)境中,RUNX2高表達于小鼠乳腺癌4T1細胞中,高表達的RUNX2可能是通過與HIF-1α的相互作用而抑制腫瘤細胞的凋亡。
[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 鹵0.24)%vs(9.3 鹵0.55; hypoxia: 19.77 鹵0.59)%vs(15.13 鹵0.32], while overexpression of RUNX2 resulted in a decrease of 4T1 cell apoptosis rate [normoxic 0.24)%vs(9.3 9.97 鹵0.27)%vs(14.07 鹵0.80 P0.05; hypoxia: 22.43 鹵1.02)%vs(34.93 鹵0.71]. Under hypoxia condition, the expression of hypoxia inducible factor 1 偽 hypoxia induced factor-1 偽 (HIF-1 偽) and RUNX2 increased. RUNX2 could form complex with HIF-1 偽. 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 偽.
【作者單位】： 第二軍醫(yī)大學免疫學研究所暨醫(yī)學免疫學國家重點實驗室;
【基金】：國家高技術研究發(fā)展計劃(863計劃)課題資助項目(No.SS2014AA020801)~~
【分類號】：R392.12

【參考文獻】

相關期刊論文 前1條

1 林Y，

本文編號：1968532