Cesar Maria Carolina Mariano, de Sant'ana Agnes Kobayashi Calvo, Mortara Renato Arruda, Souza Victória Santos, Paschoalin Thaysa, Soufen Marco Antônio, Anastácio Juliana Machado, Macedo Erenildo F, da Cunha Fernanda Fernandes Miranda, Tada Dayane Batista, Arruda Denise Costa
Núcleo Integrado de Biotecnologia, Universidade de Mogi das Cruzes- Mogi das Cruzes, Mogi das Cruzes, SP, Brazil.
College of Engineering, Bioengineering Department, Temple University, Philadelphia, PA, USA.
Cancer Chemother Pharmacol. 2025 Jul 2;95(1):67. doi: 10.1007/s00280-025-04790-9.
The BRN2 transcription factor controls the protein expression involved in cell motility and is overexpressed in melanoma. Gene mutations involved in cell signaling pathways lead to BRN2 overexpression, tumor formation and metastasis. Peptides derived from the DNA binding domain of transcription factors can compete for the transcription binding and regulate protein expression. In this work, the antitumor activity in vitro and in vivo of the peptide E24G, derived from the DNA-binding POU domain of the BRN2 transcription factor was investigated. This peptide was fragmented into two smaller peptides E12F and A12G, their antitumor activities were characterized and compared with E24G. The E24G at 1 mM significantly reduced cell motility in vitro of B16F10-Nex2 melanoma cells. E12F peptide also inhibited cell motility at a concentration eight times smaller than E24G in murine and human melanoma cells. We observed that the antitumor activity of both E24G and E12F peptides depends on the macropinocytosis displayed by tumor cells. Also, the E24G and E12F peptides induced an increase of the CDH13 expression in 50%, however the treatment with E12F increased the expression already after 12 h by 100%. In vivo assays showed that both peptides reduced the development of metastatic lung nodules without presenting toxicity to normal organs. Our results indicate that E12F and E24G peptides can restore normal expression of BRN2 target genes at the molecular level, inhibiting the cell motility. In addition, we confirmed that the peptide binds to the DNA binding site of the BRN2 transcription factor. Further studies will elucidate their mechanisms of antitumor activity, so far our results pointed out the potential application of E12F and E24G peptides as innovative treatments for metastatic melanoma.
BRN2转录因子控制参与细胞运动的蛋白质表达，且在黑色素瘤中过表达。涉及细胞信号通路的基因突变会导致BRN2过表达、肿瘤形成和转移。源自转录因子DNA结合域的肽可竞争转录结合并调节蛋白质表达。在本研究中，对源自BRN2转录因子DNA结合POU域的肽E24G的体外和体内抗肿瘤活性进行了研究。该肽被切割成两个较小的肽E12F和A12G，对它们的抗肿瘤活性进行了表征，并与E24G进行了比较。1 mM的E24G显著降低了B16F10-Nex2黑色素瘤细胞的体外细胞运动。E12F肽在鼠类和人类黑色素瘤细胞中抑制细胞运动的浓度比E24G小八倍。我们观察到E24G和E12F肽的抗肿瘤活性均取决于肿瘤细胞表现出的巨胞饮作用。此外，E24G和E12F肽使CDH13表达增加了50%，然而用E12F处理12小时后表达就增加了100%。体内试验表明，这两种肽均减少了转移性肺结节的形成，且对正常器官无毒性。我们的结果表明，E12F和E24G肽可在分子水平恢复BRN2靶基因的正常表达，抑制细胞运动。此外，我们证实该肽与BRN2转录因子的DNA结合位点结合。进一步的研究将阐明它们的抗肿瘤活性机制，目前我们的结果指出了E12F和E24G肽作为转移性黑色素瘤创新治疗方法的潜在应用。
