Jemaà Mohamed, Setti Boubaker Nouha, Kerkeni Nesrine, M Huber Stephan
Human Genetics Laboratory LR99ES10, Faculty of Medicine of Tunis, Tunis El Manar University, Tunis 2092, Tunisia.
Neurophysiology, Cellular Physiopathology and Valorisation of Biomolecules Laboratory LR18ES03, Faculty of Sciences of Tunis, Tunis El Manar University, Tunis 2092, Tunisia.
Int J Mol Sci. 2025 Jan 30;26(3):1209. doi: 10.3390/ijms26031209.
Tetraploidy is a condition in which the entire set of chromosomes doubles, most often due to errors during cell division. Tetraploidy can lead to genomic instability and significant consequences, in particular metastasis and treatment failure in tumours, including radiotherapy. The development of new strategies to sensitise these cells to treatment is of great importance. In our study, we investigated the in vitro combination of chemical treatment with the kinase inhibitor SP600125 and irradiation on diploid versus metastatic tetraploid RKO colon cancer clones. We assessed mitochondrial transmembrane potential, cell cycle and subG1 population by flow cytometry and performed clonogenic assays to evaluate cell sensitivity. We found that the combination overcomes irradiation resistance in metastatic tetraploid clones. To identify the main pathway involved in cell sensitivity, we screened the Harvard Medical School KINOMEscan library and performed a gene ontology biological process analysis. We found that the major kinases inhibited by SP600125 were ANKK1, BIKE, IKKA, JNK1, MP2K3, MP2K4, MKNK2, MYLK, PLK4, RPS6KA4(Kin,Dom,1), MYLK4 and TTK, and the pathways involved in clone sensitivity were DNA damage repair, radiation resistance and apoptosis, through JNK pathway inhibition. Finally, our main finding was that combined treatment with SP600125 and radiotherapy reduced the resistance of metastatic tetraploid cells to treatment, essentially by inhibiting the JNK pathway. This result supports a promising anti-cancer strategy to overcome the resistance of tetraploid cancer cells to irradiation.
四倍体是指整个染色体组加倍的一种情况，最常见的原因是细胞分裂过程中出现错误。四倍体可导致基因组不稳定并产生重大后果，特别是在肿瘤中会导致转移和治疗失败，包括放射治疗。开发使这些细胞对治疗敏感的新策略非常重要。在我们的研究中，我们研究了化学治疗与激酶抑制剂SP600125以及对二倍体和转移性四倍体RKO结肠癌克隆进行照射的体外联合作用。我们通过流式细胞术评估线粒体跨膜电位、细胞周期和亚G1期细胞群，并进行克隆形成试验以评估细胞敏感性。我们发现这种联合作用克服了转移性四倍体克隆的辐射抗性。为了确定参与细胞敏感性的主要途径，我们筛选了哈佛医学院的激酶组扫描文库并进行了基因本体生物学过程分析。我们发现SP600125抑制的主要激酶有ANKK1、BIKE、IKKA、JNK1、MP2K3、MP2K4、MKNK2、MYLK、PLK4、RPS6KA4（激酶、结构域、1）、MYLK4和TTK，并且通过抑制JNK途径，参与克隆敏感性的途径有DNA损伤修复、辐射抗性和细胞凋亡。最后，我们的主要发现是，SP600125与放射治疗联合使用可降低转移性四倍体细胞的治疗抗性，主要是通过抑制JNK途径。这一结果支持了一种有前景的抗癌策略，以克服四倍体癌细胞对辐射的抗性。
