PO.TB07.03 · 肿瘤生物学
通过优化的方案从新鲜前列腺切除组织建立新型、具代表性的前列腺癌细胞系
Establishing novel, representative prostate cancer cell lines from fresh prostatectomy tissues via an optimized protocol
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：现有的前列腺癌（PCa）细胞系组合通常建立自转移部位或经过大量传代，往往无法充分再现原发性前列腺肿瘤的遗传异质性及其对肿瘤微环境的依赖性。此外，迫切需要多样化、特定人群的细胞系，以研究患者结局和治疗反应差异背后的生物学因素。为填补这一关键的资源空白，我们优化并验证了一套高效方案，用于从新鲜手术标本中从头建立患者来源的原发性PCa细胞系。
方法：患者按照VCU IRB方案#20210623签署知情同意，其自我报告的种族被记录为白人（W）或非裔美国人（AA）。从36份根治性前列腺切除标本中以穿刺活检方式获取新鲜组织。每份标本产生1-3份活检样本。H&E切片由持证病理学家分级。基于Maitland等人2016年的方案进行优化，组织样本经酶消化后通过差速离心分离为不同的细胞群，并在适合上皮和成纤维细胞系的最佳培养基中扩增。通过细胞化学确认患者配对细胞系的细胞来源，并进一步通过增殖实验、Western blot和短串联重复序列（STR）分型进行表征。
结果：在试验后四分之一阶段实施的优化方案表现出高效力，成功率达86%（15份样本中的13份），使从18名患者成功建立的新型、配对的PCa上皮和成纤维细胞系总数达到30个。在这30个样本中，18个为良性（11个W和7个AA），10个为癌性（6个W和4个AA）。此外，3份患者前列腺标本同时产生了来自肿瘤和对侧非恶性组织的配对细胞系。至关重要的是，这些细胞系保留了雄激素受体（AR）和前列腺特异性抗原（PSA）的表达。此外，这些细胞系表现出异质性的生长模式，反映了源肿瘤的多样性。
结论：我们建立了一套稳健、高效的方案，用于直接从前列腺切除组织中生成患者来源的原发性PCa细胞系。这些新型、经过表征的模型显著丰富了转化研究工具箱，为多项关键应用提供了宝贵资源，包括生物标志物鉴定、理解肿瘤起始与进展、耐药机制研究、药物发现与验证，以及——最重要的是——研究特定人群差异，以指导真正公平和个体化治疗策略的开发。
查看英文原文 English abstract
Background: The existing panel of prostate cancer (PCa) cell lines, often established from metastatic sites or after extensive passaging, frequently fails to fully recapitulate the genetic heterogeneity and tumor microenvironment dependence of primary prostate tumors. Additionally, there is a crucial need for diverse, population-specific cell lines to investigate the biological factors underlying disparities in patient outcomes and therapeutic response. To address this critical resource gap, we optimized and validated a highly efficient protocol for the de novo establishment of patient-derived, primary PCa cell lines from fresh surgical specimens.
Methods: Patients were consented in accordance with VCU IRB protocol #20210623, and their self-reported race was recorded as either White (W) or African American (AA). Fresh tissues were obtained as punched biopsies from 36 radical prostatectomy specimens. Each specimen yielded 1-3 biopsy samples. H&E slides were graded by a licensed pathologist. Optimized from Maitland et al. 2016 protocol, tissue samples digested enzymatically were separated into distinct populations through differential centrifugation, and propagated in optimal media for epithelial and fibroblastic cell lines. The cellular origin of patient-matched lines were confirmed using cytochemistry and further characterized through proliferation assays, Western blot, and short tandem repeat (STR) profiling.
Results: The optimized protocol, carried out in the latter quarter of the trial, demonstrated high efficacy with an 86% success rate (13 out of 15 samples), bringing the total to 30 novel, matched PCa epithelial and fibroblast cell lines successfully established from 18 patients. Among the 30 samples, 18 were benign (11 W and 7 AA) and 10 were cancerous (6 W and 4 AA). Additionally, 3 patient prostatic specimens yielded matched cell lines from both the tumor and the contralateral non-malignant tissue. Critically, these cell lines retained the expression of androgen receptor (AR) and prostate-specific antigen (PSA). Furthermore, the lines exhibited heterogeneous growth patterns reflecting the diversity of the source tumors.
Conclusion: We have established a robust, highly effective protocol for generating patient-derived, primary PCa cell lines directly from prostatectomy tissues. These novel, characterized models significantly enrich the translational research toolkit, providing invaluable resources for several critical applications, including biomarker identification, understanding tumor initiation and progression, resistance mechanism studies, drug discovery and validation, and, most importantly, for studying population-specific differences to guide the development of truly equitable and personalized treatment strategies.
利益披露 Disclosure
T. V. Nguyen, None..
S. C. Smith, None..
L. J. Hampton, None..
V. J. Findlay, None..
D. P. Turner, None.