KLF5 and NFYA factors as novel regulators of prostate cancer cell metabolism.
KLF5 and NFYA factors as novel regulators of prostate cancer cell metabolism.
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KLF5 和 NFYA 因子作为前列腺癌细胞代谢的新型调节因子。
作者：
Raghavendra Tk Poluri;Virginie Paquette;Eric P. Allain;Camille Lafront;Charles Joly;Cindy Weidmann;A. Droit;C. Guillemette;M. Pelletier;É. Audet
前列腺癌（PCa）细胞依赖于雄激素受体（AR）信号传导轴来重新编程代谢以维持异常增殖。另外的转录因子是否参与这种重编程仍然是未知的。为了确定这些因素，在前列腺癌细胞中对雄激素敏感的基因的启动子和调控区进行DNA基序分析。这些分析确定了两个转录因子，KLF 5和NFYA，可能与前列腺癌细胞代谢。在临床数据集中，KLF 5和NFYA表达水平与疾病侵袭性相关，在PCa进展期间分别显著降低和升高。接下来在人PCa细胞模型中通过qPCR和Western印迹研究它们的表达，揭示了雄激素对KLF 5的正调控以及NFYA和AR蛋白表达状态之间的相关性。在AR阳性细胞模型中，siRNA介导的KLF 5敲低增加了人PCa细胞的增殖率，表明了肿瘤抑制功能。活细胞代谢分析表明，KLF 5的敲低促进了线粒体呼吸，这是与PCa进展相关的关键代谢途径。对于NFYA的敲低，观察到相反的增殖和呼吸，指示致癌特征。敲低KLF 5和NFYA后的RNA-seq分析证实，这两种因子调节不同的代谢基因签名以及其他基因签名，解释了它们对PCa细胞增殖和代谢的差异影响。总体而言，我们的研究结果确定KLF 5和NFYA作为PCa细胞代谢的新调节剂。
Prostate cancer (PCa) cells rely on the androgen receptor (AR) signaling axis to reprogram metabolism to sustain aberrant proliferation. Whether additional transcription factors participate to this reprogramming remains mostly unknown. To identify such factors, DNA motif analyses were performed in the promoter and regulatory regions of genes sensitive to androgens in PCa cells. These analyses identified two transcription factors, KLF5 and NFYA, as possibly associated with PCa cell metabolism. In clinical datasets, KLF5 and NFYA expression levels were associated with disease aggressiveness, being significantly decreased and increased, respectively, during PCa progression. Their expression was next investigated by qPCR and Western blot in human PCa cell models, revealing a positive regulation of KLF5 by androgens and a correlation between NFYA and AR protein expression status. siRNA-mediated knockdown of KLF5 increased human PCa cell proliferation rates in an AR-positive cell models, suggesting a tumor suppressor function. Live-cell metabolic assays showed that knockdown of KLF5 promoted mitochondrial respiration, a key metabolic pathway associated with PCa progression. The opposite was observed for knockdown of NFYA regarding proliferation and respiration, indicative of oncogenic characteristics. RNA-seq analyses following knockdown of either KLF5 and NFYA confirmed that both factors regulated distinct metabolic gene signatures, as well as other gene signatures, explaining their differential impact on PCa cell proliferation and metabolism. Overall, our findings identify KLF5 and NFYA as novel regulators of PCa cell metabolism.