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龙海晨
(2022-06-10 12:26):
#paper Ropio J, Chebly A, Ferrer J, Prochazkova-Carlotti M, Idrissi Y, Azzi-Martin L, Cappellen D, Pham-Ledard A, Soares P, Merlio JP, Chevret E. Reliable blood cancer cells' telomere length evaluation by qPCR. Cancer Med. 2020 May;9(9):3153-3162. doi: 10.1002/cam4.2816. Epub 2020 Mar 6. PMID: 32142223; PMCID: PMC7196062.题目：Reliable blood cancer cells' telomere length evaluation by qPCR
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种关联。在众多端粒长度测量方法中，qPCR被认为是一种易于进行且经济高效的方法，可以研究DNA含量较低的样本。文章采用采用相对和绝对qPCR方法检测癌细胞端粒长度。qPCR数据与terminal restriction fragment (TRF)，（端粒长度测量“金标准”）测量值进行比较。从每个样品（细胞系、患者和健康供体）的重复反应中收集数据。当重复中 Ct 的标准偏差低于 0.5 时，接受重复值。qPCR 估计的平均细胞系端粒长度（4.320 ± 0.143 kb）与 TRF 估计的（5.652 kb）.相似，P = 0.5040。对基于 qPCR 的技术，测量端粒长度要求使用高质量的 DNA，当DNA降解严重时，QPCR的测量不准确。如果QPCR法测量端粒长度的细胞是癌细胞，那关键点在于选择合适的参考基因，因为癌细胞的增殖不像正常细胞那样规范，癌细胞增殖时有些基因会丢失，癌细胞还会积累遗传和染色体异常。
Abstract:
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, …
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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.
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