Targeting breast cancer metabolism with a novel inhibitor of mitochondrial ATP synthesis.
Targeting breast cancer metabolism with a novel inhibitor of mitochondrial ATP synthesis.
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使用线粒体 ATP 合成的新型抑制剂来靶向乳腺癌代谢。
DOI：
10.18632/oncotarget.27743
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发表时间：
2020-10-27
期刊：
影响因子：
--
通讯作者：
Passaniti A
中科院分区：
文献类型：
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作者：
Kim MS;Gernapudi R;Cedeño YC;Polster BM;Martinez R;Shapiro P;Kesari S;Nurmemmedov E;Passaniti A
线粒体呼吸和ATP合成的抑制剂可以促进选择性杀死对肿瘤进展至关重要的呼吸活性癌细胞。我们以前报道过，CADD 522，一种RUNX 2转录因子的小分子抑制剂，具有治疗乳腺癌的潜力。在目前的研究中，我们发现CADD 522通过以RUNX 2独立的方式降低人乳腺癌细胞中的线粒体耗氧率（OCR）和ATP产生来抑制线粒体氧化磷酸化。CADD 522处理可抑制线粒体ATP合成酶的活性。重要的是，检测药物诱导的蛋白质稳定性的细胞热位移试验结果显示，CADD 522与F1-ATP合酶复合物的α和β亚基相互作用。差示扫描荧光法也证明了F1-ATP合酶的α亚基与CADD 522的相互作用。这些结果表明，CADD 522可能靶向ATP合酶复合物中的酶F1亚基。CADD 522增加了细胞内活性氧（ROS）的水平，这是由MitoQ（一种靶向抗氧化剂）阻止的，这表明暴露于CADD 522的癌细胞可能会升高线粒体中的ROS。外源添加的促氧化剂（例如过氧化氢或叔丁基过氧化氢）可以增强CADD 522增加的线粒体活性氧水平。相反，CADD 522介导的细胞生长抑制被N-乙酰基-L-半胱氨酸（一种一般的ROS清除剂）阻断。因此，CADD 522可能通过增加线粒体驱动的细胞ROS水平来发挥其抗肿瘤活性。总的来说，我们的数据表明，体外概念验证支持抑制线粒体ATP合酶和ROS生成是CADD 522抑制肿瘤生长的有效性的贡献者。
Inhibitors of mitochondrial respiration and ATP synthesis may promote the selective killing of respiration-competent cancer cells that are critical for tumor progression. We previously reported that CADD522, a small molecule inhibitor of the RUNX2 transcription factor, has potential for breast cancer treatment. In the current study, we show that CADD522 inhibits mitochondrial oxidative phosphorylation by decreasing the mitochondrial oxygen consumption rate (OCR) and ATP production in human breast cancer cells in a RUNX2-independent manner. The enzyme activity of mitochondrial ATP synthase was inhibited by CADD522 treatment. Importantly, results from cellular thermal shift assays that detect drug-induced protein stabilization revealed that CADD522 interacts with both α and β subunits of the F1-ATP synthase complex. Differential scanning fluorimetry also demonstrated interaction of α subunits of the F1-ATP synthase to CADD522. These results suggest that CADD522 might target the enzymatic F1 subunits in the ATP synthase complex. CADD522 increased the levels of intracellular reactive oxygen species (ROS), which was prevented by MitoQ, a mitochondria-targeted antioxidant, suggesting that cancer cells exposed to CADD522 may elevate ROS from mitochondria. CADD522-increased mitochondrial ROS levels were enhanced by exogenously added pro-oxidants such as hydrogen peroxide or tert-butyl hydroperoxide. Conversely, CADD522-mediated cell growth inhibition was blocked by N-acetyl-l-cysteine, a general ROS scavenger. Therefore, CADD522 may exert its antitumor activity by increasing mitochondrial driven cellular ROS levels. Collectively, our data suggest in vitro proof-of-concept that supports inhibition of mitochondrial ATP synthase and ROS generation as contributors to the effectiveness of CADD522 in suppression of tumor growth.