Reliable blood cancer cells' telomere length evaluation by qPCR

Background: 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. Methods: Cancer cells’ telomere length was evaluated by relative and absolute qPCR methods. Results: 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). Conclusions: 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.This work was sponsored by grants from the French Society of Dermatology (SFD), the Cancer League Committee of Dordogne, and the ARC foundation for cancer research. Joana Ropio was supported by grants from Programme Hubert Curien PESSOA-FCT, Programme d’Actions Universitaires Integrees Luso-Francaises (PAUILF) and ERASMUS+. Further funding was obtained from the project “Advancing cancer research: from basic knowledge to application” NORTE-01-0145-FEDER-000029: “Projetos Estruturados de I & D & I”, funded by Norte 2020—Programa OperacionalRegional do Norte and the project PTDC/MED-ONC/31438/2017 (The other faces of Telomerase: Looking beyond tumorimmortalization), supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF), COMPETE 2020 – Operacional Programme for Competitiveness and Internationalization (POCI) and by Portuguese funds through FCT – Fundação para a Ciência e a Tecnologia/Ministério da Ciência, Tecnologia e Ensino Superior