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Quantitative detection of disseminated melanoma cells by trp-1 transcript analysis reveals stochastic distribution of pulmonary metastases

Kyjacova, L.; Saup, R.; Rothley, M.; Schmaus, A.; Wagner, T.; Boßerhoff, A.; Garvalov, B. K.; Thiele, W.; Sleeman, J. P.


A better understanding of the process of melanoma metastasis is required to underpin the development of novel therapies that will improve patient outcomes. The use of appropriate animal models is indispensable for investigating the mechanisms of melanoma metastasis. However, reliable and practicable quantification of metastases in experimental mice remains a challenge, particularly if the metastatic burden is low. Here, we describe a qRT-PCR-based protocol that employs the melanocytic marker Trp-1 for the sensitive quantification of melanoma metastases in the murine lung. Using this protocol, we were able to detect the presence of as few as 100 disseminated melanoma cells in lung tissue. This allowed us to quantify metastatic burden in a spontaneous syngeneic B16-F10 metastasis model, even in the absence of visible metastases, as well as in the autochthonous Tg(Grm1)/Cyld$^{-/-}$ melanoma model. Importantly, we also observed an uneven distribution of disseminated melanoma cells amongst the five lobes of the murine lung, which varied considerably from animal to animal. Together, our findings demonstrate that the qRT-PCR-based detection of Trp-1 allows the quantification of low pulmonary metastatic burden in both transplantable and autochthonous murine melanoma models, and show that the analysis of lung metastasis in such models needs to take into account the stochastic distribution of metastatic lesions amongst the lung lobes.

Verlagsausgabe §
DOI: 10.5445/IR/1000140526
Veröffentlicht am 03.12.2021
DOI: 10.3390/jcm10225459
Zitationen: 2
Web of Science
Zitationen: 2
Zitationen: 2
Cover der Publikation
Zugehörige Institution(en) am KIT Institut für Biologische und Chemische Systeme (IBCS)
Publikationstyp Zeitschriftenaufsatz
Publikationsjahr 2021
Sprache Englisch
Identifikator ISSN: 2077-0383
KITopen-ID: 1000140526
HGF-Programm 47.14.02 (POF IV, LK 01) Information Storage and Processing in the Cell Nucleus
Erschienen in Journal of Clinical Medicine
Verlag MDPI
Band 10
Heft 22
Seiten Art.Nr. 5459
Vorab online veröffentlicht am 22.11.2021
Nachgewiesen in Dimensions
Web of Science
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