Stulpinas Aurimas, Sereika Matas, Vitkeviciene Aida, Imbrasaite Ausra, Krestnikova Natalija, Kalvelyte Audrone V
Department of Molecular Cell Biology, Institute of Biochemistry, Life Sciences Center, Vilnius University, Vilnius, Lithuania.
Front Oncol. 2023 Jan 4;12:1045521. doi: 10.3389/fonc.2022.1045521. eCollection 2022.
There is no doubt that cell signaling manipulation is a key strategy for anticancer therapy. Furthermore, cell state determines drug response. Thus, establishing the relationship between cell state and therapeutic sensitivity is essential for the development of cancer therapies. In the era of personalized medicine, the use of patient-derived ex vivo cell models is a promising approach in the translation of key research findings into clinics. Here, we were focused on the non-oncogene dependencies of cell resistance to anticancer treatments. Signaling-related mechanisms of response to inhibitors of MEK/ERK and PI3K/AKT pathways (regulators of key cellular functions) were investigated using a panel of patients' lung tumor-derived cell lines with various stemness- and EMT-related markers, varying degrees of ERK1/2 and AKT phosphorylation, and response to anticancer treatment. The study of interactions between kinases was the goal of our research. Although MEK/ERK and PI3K/AKT interactions are thought to be cell line-specific, where oncogenic mutations have a decisive role, we demonstrated negative feedback loops between MEK/ERK and PI3K/AKT signaling pathways in all cell lines studied, regardless of genotype and phenotype differences. Our work showed that various and distinct inhibitors of ERK signaling - selumetinib, trametinib, and SCH772984 - increased AKT phosphorylation, and conversely, inhibitors of AKT - capivasertib, idelalisib, and AKT inhibitor VIII - increased ERK phosphorylation in both control and cisplatin-treated cells. Interaction between kinases, however, was dependent on cellular state. The feedback between ERK and AKT was attenuated by the focal adhesion kinase inhibitor PF573228, and in cells grown in suspension, showing the possible role of extracellular contacts in the regulation of crosstalk between kinases. Moreover, studies have shown that the interplay between MEK/ERK and PI3K/AKT signaling pathways may be dependent on the strength of the chemotherapeutic stimulus. The study highlights the importance of spatial location of the cells and the strength of the treatment during anticancer therapy.
毫无疑问,细胞信号传导调控是抗癌治疗的关键策略。此外,细胞状态决定药物反应。因此,建立细胞状态与治疗敏感性之间的关系对于癌症治疗的发展至关重要。在个性化医疗时代,使用患者来源的体外细胞模型是将关键研究成果转化为临床应用的一种有前景的方法。在此,我们聚焦于细胞对抗癌治疗耐药的非癌基因依赖性。我们使用一组具有各种干性和上皮-间质转化(EMT)相关标志物、不同程度的ERK1/2和AKT磷酸化以及对抗癌治疗反应的患者肺肿瘤衍生细胞系,研究了对MEK/ERK和PI3K/AKT通路(关键细胞功能的调节因子)抑制剂的信号相关反应机制。激酶之间相互作用的研究是我们的研究目标。尽管MEK/ERK和PI3K/AKT的相互作用被认为是细胞系特异性的,其中致癌突变起决定性作用,但我们证明了在所有研究的细胞系中,无论基因型和表型差异如何,MEK/ERK和PI3K/AKT信号通路之间都存在负反馈回路。我们的工作表明,各种不同的ERK信号抑制剂——司美替尼、曲美替尼和SCH772984——增加了AKT磷酸化,相反,AKT抑制剂——卡匹西他赛、idelalisib和AKT抑制剂VIII——在对照细胞和顺铂处理细胞中均增加了ERK磷酸化。然而,激酶之间的相互作用取决于细胞状态。粘着斑激酶抑制剂PF573228减弱了ERK和AKT之间的反馈,并且在悬浮培养的细胞中也是如此,这表明细胞外接触在调节激酶间串扰中可能发挥的作用。此外,研究表明MEK/ERK和PI3K/AKT信号通路之间的相互作用可能取决于化疗刺激的强度。该研究突出了抗癌治疗期间细胞空间位置和治疗强度的重要性。