本文選題：乳腺癌細(xì)胞 + Runt相關(guān)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(guān)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(xì)胞凋亡的影響及其作用機(jī)制。方法:Real-time PCR和Western blotting分別檢測低氧條件對4T1細(xì)胞內(nèi)RUNX1、RUNX2、RUNX3 mRNA和RUNX2蛋白表達(dá)的影響;采用小干擾RNA技術(shù)與真核重組質(zhì)粒DNA過表達(dá)技術(shù)降低或者提高4T1細(xì)胞RUNX2的表達(dá),免疫共沉淀法檢測4T1細(xì)胞中RUNX2與其他蛋白之間的相互結(jié)合情況,流式細(xì)胞術(shù)檢測常氧/低氧條件下干擾/過表達(dá)RUNX2對4T1細(xì)胞凋亡的影響。結(jié)果:低氧條件下4T1細(xì)胞中RUNX1和RUNX2 mRNA的表達(dá)水平上調(diào)(P0.05),RUNX2的蛋白表達(dá)量明顯增加;轉(zhuǎn)染RUNX2-siRNA-1415和真核表達(dá)載體pc DNA3.1(-)-RUNX2可使4T1細(xì)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(dá)均導(dǎo)致小鼠乳腺癌4T1細(xì)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(dá)RUNX2均導(dǎo)致4T1細(xì)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(dǎo)因子-1α(hypoxia induced factor-1α,HIF-1α)與RUNX2的表達(dá)上升,RUNX2能與HIF-1α形成復(fù)合物。結(jié)論:在腫瘤低氧微環(huán)境中,RUNX2高表達(dá)于小鼠乳腺癌4T1細(xì)胞中,高表達(dá)的RUNX2可能是通過與HIF-1α的相互作用而抑制腫瘤細(xì)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(xué)免疫學(xué)研究所暨醫(yī)學(xué)免疫學(xué)國家重點(diǎn)實(shí)驗(yàn)室;
【基金】：國家高技術(shù)研究發(fā)展計(jì)劃(863計(jì)劃)課題資助項(xiàng)目(No.SS2014AA020801)~~
【分類號】：R392.12

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