32-qter (17%) was observable in all SB431542 mouse passages of a series (case number 445). The genomic region 1q21.1-qter frequently displayed gain. Changes in copy number were acquired during the growth period of xenografts including gains at 2q35-q37.3, 4q13.3-qter, 8p11.21-p21.2 and 8q and losses at 8p, 17p, 13, Xq21.1, 1p13.3-p31.1, 5q, 11q13.4-q24.3, Xq12-q26.3 and 16q. In one xenograft GSK2126458 research buy series (Case number 488), loss of 17q12-q21.32, that was present in the early passages, disappeared

during the growth process. The loss of 1p36.12-pter in the first two passages originating from lung metastasis (1 and 4) changed to loss of 1p36.21-pter in the last three passages (14, 21, and 30). The lung metastasis xenografts showed 9 copy number changes, whereas only 3 of these aberrations were observable in the xenograft passages from its primary tumor. Table 2 The copy number changes present in all the passages of each xenograft series Case No. (Nude) Array CGH results 488 (15) +1q21.1-qter, -13q14.12-qter

445 (22) -2q35-q37.3 (uncontinuous), + 8, +15, +17q21.32-qter 451 (53) -1q24.3-q25.2, – 3p12.3-p24.3, -9p21.3 455 (199) +1q, -16q, -9p21.3 430 (PRI) (230) -9p21.3 430 (MET) (248) -1p36.12-pter, -9p21.3 PRI = Primary Tumor, MET = Lung Metastasis Table 3 Copy number changes present in only part of the passages of each xenograft series Case Nude- Passage Array CGH result 488 15- 2, 4, 7, 11, 14 -2q35-q37.3 488 15- 1, 2, 4, 7 -17q12-q21.32 488 15- 14 +17 451 53- 11, 15,18, 21 +4q13.3-qter, -17p 455 199- 5, 11, 17, 25 -13 455 199- 25 -Xq21.1 430 (PRI) 230- SRT1720 nmr 1, 4, 9, 19 +8p11.21-p21.2, +8q 430 (MET) 248- 1, 4 -1p36.12-pter 430 (MET) 248- 14, 21, 30 -1p36.21-pter, -1p13.3-p31.1, -5q,     -11q13.4-q24.3, -Xq12-q26.3 430 (MET) 248- 21, 30 +8p11.21-p21.2, +8q 430 (MET) 248- 30 -16q PRI = Primary Tumor, MET = Lung Metastasis Figure 1 Copy number changes on each chromosome were ordered using hierarchical clustering. Most of the

xenograft passages of each series clustered together and also with the passage 0, its corresponding primary tumor. MicroRNA alterations in xenografts Differences in miRNA expression between xenografts and control samples were detected upon analysis (Figure 2). Exclusively expressed miRNAs were detected; two in control samples filipin (miR-31, miR-31*) and 46 in all xenograft passages (Table 4). In addition, 5 miRNAs (miR-106b, miR-93, miR-181b, miR-101, miR-30b) were significantly over-expressed (q-value < 0.05), while 6 miRNAs (miR-145, miR-193a-3p, miR-100, miR-22, miR-21, miR-574-3p) were significantly under-expressed across the xenograft passages in relation to the controls (q-value < 0.05). Xenografts from primary and control samples were compared to xenograft passages from the lung metastasis (Case number 430), to determine differences in miRNA expression.