Bordeaux University, INSERM U1053 Bordeaux Research in Translational Oncology (BaRITOn), Cutaneous Lymphoma Oncogenesis Team, Bordeaux, France.
Porto University, Institute of Biomedical Sciences of Abel Salazar, Porto, Portugal.
Cancer Med. 2020 May;9(9):3153-3162. doi: 10.1002/cam4.2816. Epub 2020 Mar 6.
Telomere shortening is linked to a range of different human diseases, hence reliable measurement methods are needed to uncover such associations. Among the plethora of telomere length measurement methods, qPCR is reported as easy to conduct and a cost-effective approach to study samples with low DNA amounts.
Cancer cells' telomere length was evaluated by relative and absolute qPCR methods.
Robust and reproducible telomere length measurements were optimized taking into account a careful reference gene selection and by knowing the cancer cells ploidy. qPCR data were compared to "gold standard" measurement from terminal restriction fragment (TRF).
Our study provides guidance and recommendations for accurate telomere length measurement by qPCR in cancer cells, taking advantage of our expertise in telomere homeostasis investigation in primary cutaneous T-cell lymphomas. Furthermore, our data emphasize the requirement of samples with both, high DNA quality and high tumor cells representation.
端粒缩短与一系列不同的人类疾病有关，因此需要可靠的测量方法来揭示这些关联。在众多端粒长度测量方法中，qPCR 被报道为一种易于进行且具有成本效益的方法，可用于研究 DNA 量低的样本。
通过相对和绝对 qPCR 方法评估癌细胞的端粒长度。
考虑到精心选择的参考基因和癌细胞的倍性，优化了稳健且可重复的端粒长度测量。qPCR 数据与端粒限制片段（TRF）的“金标准”测量进行了比较。
我们的研究利用我们在原发性皮肤 T 细胞淋巴瘤中端粒动态平衡研究方面的专业知识，为癌症细胞中通过 qPCR 进行准确的端粒长度测量提供了指导和建议。此外，我们的数据强调了需要具有高质量 DNA 和高肿瘤细胞代表性的样本。
