Saydullaeva Iroda, Butuner Bilge Debelec, Korkmaz Kemal Sami
Faculty of Engineering, Department of Bioengineering, Cancer Biology Laboratory, Ege University, Izmir 35040, Turkey.
Faculty of Pharmacy, Department of Pharmaceutical Biotechnology, Ege University, Izmir 35040, Turkey.
ACS Omega. 2023 Sep 1;8(36):32580-32592. doi: 10.1021/acsomega.3c03127. eCollection 2023 Sep 12.
Studies demonstrate that inflammation synergizes with high-grade aggressive prostate tumor development and ultimately metastatic spread, in which a lot of work has been done in recent years. However, the clear mechanism of inflammation inciting prostate cancer remains largely uncharacterized. Our previous study has shown that the conditioned media (CM)-mediated LNCaP cell migration is partially correlated with the loss of expression of the tumor suppressor NKX3.1. Here, we continue to investigate the inflammation-mediated migration of prostate cancer cells, and the role of NKX3.1 in this process to gain insights into cell migration-related changes comprehensively. Earlier, the model of inflammation in the tumor microenvironment have been optimized by our research group; here, we continue to investigate the time-dependent effect of CM exposure together with NKX3.1 changes, in which we observed that these changes play important roles in gaining heterogeneous epithelial-to-mesenchymal transition (EMT) phenotype. Hence, this is an important parameter of tumor progression; we depleted NKX3.1 expression using the CRISPR/Cas9 system and examined the migrating cell clusters after exposure to inflammatory cytokines. We found that the migrated cells clearly demonstrate reversible loss of E-cadherin expression, which is consistent with subsequent vimentin expression alterations in comparison to control cells. Moreover, the data suggest that the AR-mediated transcriptional program also contributes to mesenchymal-to-epithelial transition (MET) in prostate cancer progression. Furthermore, the quantitative proteomic analysis showed that migrated subpopulations from the same cell line presented different phenotypes in which the proteins overexpressed are involved in cell metabolism and RNA processing. According to KEGG pathway analysis, the ABC transporters were found to be the most significant. Thus, the dynamic process of cellular migration favors diverse genetic compositions under changing tumor microenvironments. The different levels of invasiveness are supported by shifting the cells in between these EMT and MET phenotypes.
研究表明,炎症与高级别侵袭性前列腺肿瘤的发展以及最终的转移扩散相互协同,近年来对此已开展了大量研究工作。然而,炎症引发前列腺癌的明确机制在很大程度上仍未明确。我们之前的研究表明,条件培养基(CM)介导的LNCaP细胞迁移与肿瘤抑制因子NKX3.1表达缺失部分相关。在此,我们继续研究炎症介导的前列腺癌细胞迁移以及NKX3.1在此过程中的作用,以全面深入了解细胞迁移相关变化。此前,我们研究小组已优化了肿瘤微环境中的炎症模型;在此,我们继续研究CM暴露的时间依赖性效应以及NKX3.1的变化,在此过程中我们观察到这些变化在获得异质性上皮-间质转化(EMT)表型中发挥重要作用。因此,这是肿瘤进展的一个重要参数;我们使用CRISPR/Cas9系统降低NKX3.1的表达,并在暴露于炎性细胞因子后检测迁移的细胞簇。我们发现,迁移的细胞明显表现出E-钙黏蛋白表达的可逆性缺失,与对照细胞相比,这与随后波形蛋白表达的改变一致。此外,数据表明,雄激素受体(AR)介导的转录程序也有助于前列腺癌进展过程中的间质-上皮转化(MET)。此外,定量蛋白质组学分析表明,来自同一细胞系的迁移亚群呈现出不同的表型,其中过表达的蛋白质参与细胞代谢和RNA加工。根据京都基因与基因组百科全书(KEGG)通路分析,发现ABC转运蛋白最为显著。因此,细胞迁移的动态过程有利于在不断变化的肿瘤微环境下形成多样化的基因组成。通过在这些EMT和MET表型之间转换细胞来支持不同程度的侵袭性。