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Analysis of Tumor Cell Evolution in a Melanoma: Evidence of Mutational and Selective Pressure for Loss of p16ink4 and for Microsatellite Instability Albert Rübben, Philipp Babilas, Jens M. Baron, Anja Hofheinz, Mark Neis, Florian Sels, Markus Sporkert Journal of Investigative Dermatology Volume 114, Issue 1, Pages (January 2000) DOI: /j x Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions
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Figure 1 Competitive multiplex PCR for demonstration of loss of the p16ink4 gene. (a) Calibration of primer ratio, (b) PCR with extracted DNA from primary tumor (P1–P3), metastases (M6, M7) and blood of the patient. pBR322, molecular size marker. Journal of Investigative Dermatology , 14-20DOI: ( /j x) Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions
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Figure 2 Comparison of the methods used for the analysis of PCR-amplified microsatellite DNA. (a) TG-SSCP analysis, (b) analysis in a sequencing gel, (c) conventional sequencing gel versus Southern blot hybridization of the blotted sequencing gel with a (CA)10-oligonucleotide; M, metastases; N, normal formalin-fixed tissue specimens; ▸ loss of DNA bands (LOH); ▹ mutated DNA bands (microsatellite instability); ◃* points to nonspecific DNA bands. Journal of Investigative Dermatology , 14-20DOI: ( /j x) Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions
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Figure 3 Characterization of tumor cell populations. TG-SSCP analysis of PCR-amplified microsatellite DNA obtained from microdissected tissue of the primary tumor (P1–3), from metastases (M7, M6) and from the patient’s blood (B). ▸ loss of DNA bands (LOH); ▹ mutated DNA bands. Journal of Investigative Dermatology , 14-20DOI: ( /j x) Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions
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Figure 4 Putative genealogic tree of tumor cell evolution in the analysed patient obtained from data summarized in Table 1. Journal of Investigative Dermatology , 14-20DOI: ( /j x) Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions
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