Institute of Pathology, University Hospital Essen, University of Duisburg Essen, 45147 Essen, Germany.
Department of Pulmonary Medicine, University Hospital Essen-Ruhrlandklinik, University of Duisburg Essen, 45147 Essen, Germany.
Int J Mol Sci. 2022 Mar 18;23(6):3278. doi: 10.3390/ijms23063278.
Malignant pleural mesothelioma (MPM) has an infaust prognosis due to resistance to systemic treatment with platin-analoga. MPM cells modulate the immune response to their benefit. They release proinflammatory cytokines, such as TGF-ß, awakening resting fibrocytes that switch their phenotype into activated fibroblasts. Signaling interactions between cancer cells and cancer-associated fibroblasts (CAFs) play an integral part in tumor progression. This study aimed to investigate the role CAFs play in MPM progression, analyzing the impact this complex, symbiotic interaction has on kinase-related cell signaling in vitro.
We simulated paracrine signaling in vitro by treating MPM cell lines with conditioned medium (CM) from fibroblasts (FB) and vice versa. NCI-H2052, MSTO-211H, and NCI-H2452 cell lines representing the three mayor MPM subtypes, while embryonal myofibroblast cell lines, IMR-90 and MRC-5, provide a CAFs-like phenotype. Subsequently, differences in proliferation rates, migratory behavior, apoptosis, necrosis, and viability were used as covariates for data analysis. Kinase activity of treated samples and corresponding controls were then analyzed using the PamStation12 platform (PamGene); Results: Treatment with myofibroblast-derived CM revealed significant changes in phosphorylation patterns in MPM cell lines. The observed effect differs strongly between the analyzed MPM cell lines and depends on the origin of CM. Overall, a much stronger effect was observed using CM derived from IMR-90 than MRC-5. The phosphorylation changes mainly affected the MAPK signaling pathway.; Conclusions: The factors secreted by myofibroblasts in fibroblasts CM significantly influence the phosphorylation of kinases, mainly affecting the MAPK signaling cascade in tested MPM cell lines. Our in vitro results indicate promising therapeutic effects by the use of MEK or ERK inhibitors and might have synergistic effects in combination with cisplatin-based treatment, improving clinical outcomes for MPM patients.
恶性胸膜间皮瘤（MPM）由于对顺铂类似物的全身治疗产生耐药性，预后不佳。MPM 细胞调节其获益的免疫反应。它们释放促炎细胞因子，如 TGF-β，唤醒静止的成纤维细胞，将其表型转变为激活的成纤维细胞。癌细胞与癌相关成纤维细胞（CAF）之间的信号相互作用在肿瘤进展中起着至关重要的作用。本研究旨在探讨 CAF 在 MPM 进展中的作用，分析这种复杂的共生相互作用对体外激酶相关细胞信号的影响。
我们通过用成纤维细胞（FB）的条件培养基（CM）或反之处理 MPM 细胞系来模拟旁分泌信号。NCI-H2052、MSTO-211H 和 NCI-H2452 细胞系代表三种主要的 MPM 亚型，而胚胎成纤维细胞系 IMR-90 和 MRC-5 提供了 CAF 样表型。随后，将增殖率、迁移行为、凋亡、坏死和活力的差异用作数据分析的协变量。然后使用 PamStation12 平台（PamGene）分析处理样品和相应对照的激酶活性；结果：用成肌纤维细胞衍生的 CM 处理揭示了 MPM 细胞系中磷酸化模式的显著变化。观察到的效应在分析的 MPM 细胞系之间差异很大，并且取决于 CM 的来源。总体而言，使用来自 IMR-90 的 CM 观察到的效应要强得多。磷酸化变化主要影响 MAPK 信号通路；结论：成纤维细胞 CM 中肌成纤维细胞分泌的因子显著影响激酶的磷酸化，主要影响测试的 MPM 细胞系中的 MAPK 信号级联。我们的体外结果表明，使用 MEK 或 ERK 抑制剂具有有希望的治疗效果，并且可能与顺铂为基础的治疗联合具有协同作用，改善 MPM 患者的临床结果。
