Wright Jessica A, White Stephanie D, Frame Gavin, Bosiljkov Ana, Khan Shahbaz, Haas Roni, Yang Qian, Sheng Minzhi, Huang Xiaoyong, Higgins Geoff S, Mills Ian, Downes Michelle R, Vesprini Danny, Chung Hans T, Screaton Robert A, Leong Hon S, Boutros Paul C, Kislinger Thomas, Liu Stanley K
Sunnybrook Research Institute, Toronto, ON M4N 3M5, Canada.
Department of Medical Biophysics, University of Toronto, Toronto, ON M5G 2C4, Canada.
Cells. 2025 May 31;14(11):819. doi: 10.3390/cells14110819.
Prostate cancer cell lines are particularly clinically homogenous, mostly representing metastatic states rather than localized disease. While there has been significant work in the development of additional models, few have been created without oncogenic transformation. We derived a primary prostate cancer cell line from a patient with localized Gleason 7 prostate cancer-designated CaB34-which spontaneously immortalized. We leveraged CaB34 to generate a paired radioresistant subline, CaB34-CF, using a clinically relevant fractionated radiotherapy schedule. These two paired cell lines were investigated extensively to determine their molecular characteristics and therapy responses. Both CaB34 and CaB34-CF express prostate-specific markers, including KRT18, NKX3.1, and AMACR. Multi-omic analyses using RNAseq and shotgun proteomics identified NNMT as the most significantly dysregulated component in CaB34-CF. A bioinformatic analysis determined that NNMT was more abundant within prostate tumors compared to benign prostate, suggesting a role in tumor progression. Knockdown studies of NNMT demonstrated significant radiosensitization of CaB34-CF cells, which was largely a result of increased radiation-induced cellular senescence. Growth as 3D organoids was significantly higher in the CaB34-CF line, and demonstrated a less structured pattern of expression of cytokeratin markers. Radiosensitization with NNMT siRNA was recapitulated in a 3D organoid clonogenic assay in CaB34-CF cells. Our research provides a paired primary model of treatment-naïve and radioresistant disease to address mechanisms of therapy resistance, while expanding the repertoire of localized prostate cancer cell lines for the research community. In addition, our findings present NNMT as a potential therapeutic target for sensitization of radioresistant disease.
前列腺癌细胞系在临床上具有特别的同质性，大多代表转移状态而非局限性疾病。虽然在开发其他模型方面已经开展了大量工作，但很少有模型是在没有致癌转化的情况下创建的。我们从一名患有局限性Gleason 7级前列腺癌的患者身上获得了一个原发性前列腺癌细胞系，命名为CaB34，该细胞系自发永生化。我们利用CaB34，通过临床相关的分次放射治疗方案，生成了一个配对的放射抗性亚系CaB34-CF。对这两个配对的细胞系进行了广泛研究，以确定它们的分子特征和治疗反应。CaB34和CaB34-CF均表达前列腺特异性标志物，包括角蛋白18（KRT18）、NKX3.1和α-甲基酰基辅酶A消旋酶（AMACR）。使用RNA测序（RNAseq）和鸟枪法蛋白质组学进行的多组学分析确定，NNMT是CaB34-CF中失调最显著的成分。生物信息学分析确定，与良性前列腺相比，NNMT在前列腺肿瘤中含量更高，表明其在肿瘤进展中起作用。对NNMT的敲低研究表明，CaB34-CF细胞有显著的放射增敏作用，这在很大程度上是辐射诱导的细胞衰老增加的结果。CaB34-CF细胞系中三维类器官的生长明显更高，并且细胞角蛋白标志物的表达模式结构较少。在CaB34-CF细胞的三维类器官克隆形成试验中，NNMT小干扰RNA（siRNA）的放射增敏作用得到了重现。我们的研究提供了一个未经治疗的和放射抗性疾病的配对原发性模型，以解决治疗抗性机制，同时为研究界扩展局限性前列腺癌细胞系的种类。此外，我们的研究结果表明，NNMT是放射抗性疾病增敏的一个潜在治疗靶点。
