Figure 6.

Cell cycle control and tumor suppressor function are disrupted by different mechanisms in both metastatic sites. (A) Genomic regions encompassing CDKN2A, CCND1, and CCNE1 with copy-number gains or losses in lung tumor (red) and metastatic lymph node (blue) as assessed by CNV-seq analysis of WGS data. (B) Relative levels of CDKN2A, CCND1, and CCNE1 expression in the lung tumor and lymph nodes were determined by quantitative RT-PCR using POLR2A mRNA as an endogenous control. Samples were analyzed in quadruplicate, and values expressed as the mean ± SD. (C) Region encompassing TP53 with copy-number loss in the lung tumor (L-red) and focal loss of TP53 in the metastatic lymph node (LN1-blue) as assessed by CNV-seq analysis of WGS data. (D) Expression of TP53 in the lung tumor and metastatic lymph nodes was determined by quantitative reverse transcription–polymerase chain reaction (RT-PCR) with POLR2A mRNA as an endogenous control. Samples were analyzed in quadruplicate, and values expressed as the mean ± SD. (E) Chromatogram obtained from Sanger sequencing of cDNA shows that TP53 with an in-frame 9-nt somatic deletion (Del EMF) is expressed in the lung tumor, whereas wild-type TP53 is expressed in the lymph node.