对于血液系统恶性肿瘤细胞系中DNA损伤响应途径的研究是寻找新的治疗犬癌症靶点的必要步骤。
Studying the DNA damage response pathway in hematopoietic canine cancer cell lines, a necessary step for finding targets to generate new therapies to treat cancer in dogs.
发表日期：2023
作者：
Beatriz Hernández-Suárez, David A Gillespie, Ewa Dejnaka, Piotr Kupczyk, Bożena Obmińska-Mrukowicz, Aleksandra Pawlak
来源：
CLINICAL PHARMACOLOGY & THERAPEUTICS
摘要：
狗在比较肿瘤学研究中提供了重要的机会。然而，目前在犬细胞中研究癌症生物学现象受制于受限的抗体试剂和技术的可获得性。在这里，我们利用商业可获得的抗体试剂对一系列造血性犬癌细胞系的DNA损伤应答（DDR）关键成分的表达和活性进行了初步的表征。用于此验证分析的技术包括Western blot、qPCR和DNA梳状分析。在犬癌细胞系中观察到DDR成分的基础表达（ATR、Claspin、Chk1和Rad51）和激动剂诱导的活化（p-Chk1）均存在显著变异。CLBL-1（B细胞淋巴瘤）和CLB70（B细胞慢性淋巴细胞白血病）细胞系的表达比GL-1（B细胞白血病）细胞系更强，但这些差异的生物学意义需要进一步研究。我们还验证了在造血性犬癌细胞系中定量DNA复制动力学的方法，并发现GL-1细胞系的复制叉速度高于CLBL-1细胞系，但两者均显示出复制叉不对称的趋势。这些发现将对癌症生物学的未来研究提供信息，有助于发展针对狗患者的新型抗癌疗法。它们还可以为人类肿瘤学提供新的知识。
版权所有 © 2023 Hernández-Suárez, Gillespie, Dejnaka, Kupczyk, Obmińska-Mrukowicz and Pawlak.
Dogs present a significant opportunity for studies in comparative oncology. However, the study of cancer biology phenomena in canine cells is currently limited by restricted availability of validated antibody reagents and techniques. Here, we provide an initial characterization of the expression and activity of key components of the DNA Damage Response (DDR) in a panel of hematopoietic canine cancer cell lines, with the use of commercially available antibody reagents.The techniques used for this validation analysis were western blot, qPCR, and DNA combing assay.Substantial variations in both the basal expression (ATR, Claspin, Chk1, and Rad51) and agonist-induced activation (p-Chk1) of DDR components were observed in canine cancer cell lines. The expression was stronger in the CLBL-1 (B-cell lymphoma) and CLB70 (B-cell chronic lymphocytic leukemia) cell lines than in the GL-1 (B-cell leukemia) cell line, but the biological significance of these differences requires further investigation. We also validated methodologies for quantifying DNA replication dynamics in hematopoietic canine cancer cell lines, and found that the GL-1 cell line presented a higher replication fork speed than the CLBL-1 cell line, but that both showed a tendency to replication fork asymmetry.These findings will inform future studies on cancer biology, which will facilitate progress in developing novel anticancer therapies for canine patients. They can also provide new knowledge in human oncology.Copyright © 2023 Hernández-Suárez, Gillespie, Dejnaka, Kupczyk, Obmińska-Mrukowicz and Pawlak.