Supporting tables and figures.

Table A in S1 File. Culture conditions for cell lines. Difference (Δ) in passage number between late and early passage cells. Doublings were calculated based on 4.32 and 7.64 doublings at splitting ratios of 1:20 and 1:200, respectively. Table B in S1 File. Distribution of genes on chromosomes in the human and mouse arrays. n.p., not present in mouse; %, per cent of sum. Table C in S1 File. Primers used for real-time PCR analysis. Table D in S1 File. Chromosomal localization of differentially expressed genes after serial passaging and after selection for increased metastatic activity. Imbalances (p2fold; p20. Analysis for metastasis-related genes was performed on pooled data sets obtained from early and late passaged cells. n; number of affected chromosomes. Preferential up-regulation is indicated in bold. Table E in S1 File. Chromosomal macro-aberrations after serial in vitro passaging of SAOS and LM5 cells. Commonly affected regions in SAOS and LM5 are indicated in yellow. Table F in S1 File. Correlation of regulated genes after serial in vitro passaging of SAOS and LM5 cells in microarray analysis with CN gains and losses in aCGH analysis. Fig A in S1 File. Chromosomal localization of regulated genes that correlate with CN gains and losses. A) Up-regulated genes in late compared to early SAOS that correlate with CN gain. B) Down-regulated genes in late compared to early SAOS that correlate with CN loss. C) Up-regulated genes in late compared to early LM5 that correlate with CN gain. D) Down-regulated genes in late compared to early LM5 that correlate with CN loss. Table G in S1 File. Chromosomal macro-aberrations after selection for increased metastatic activity in SAOS/LM5 cell system. The most affected regions are indicated in yellow. Fig B in S1 File. Real-time PCR analysis. Comparison of commonly up-regulated genes belonging to "Focal adhesion" in late versus early passaged SAOS and LM5 cells by microarray (MA) and real-time (qPCR) analyses. Table H in S1 File. Regulated (>2-fold; p2-fold; pin vitro passaging that belong to the "Hedgehog signaling pathway" and/or "WNT signaling pathway". Comparison is late versus early passage. Genes indicated in red are also regulated after selection for increased metastatic activity (Table K in S1 File). Table J in S1 File. Enrichment of regulated genes after selection for increased metastatic potential that belong to "Hedgehog signaling pathway" and "WNT signaling pathway" depending on individual comparisons of early and late passage data sets. Metastatic/parental; HH, "Hedgehog signaling pathway"; WNT, "WNT signaling pathway"; Total, total number of regulated (>2-fold; p2-fold; p2-fold and/or pin vitro passaging in either the parental or metastatic cell line (Table H). The means are calculated for all four comparisons. Fig C in S1 File. KEGG pathways. (PDF)

补充材料1（S1 File）中的表A：细胞系培养条件。晚期传代细胞与早期传代细胞的传代次数差值（Δ）。倍增次数根据传代比例1:20和1:200时分别对应的4.32次和7.64次倍增计算得到。
补充材料1（S1 File）中的表B：人源和小鼠芯片上的基因染色体分布情况。n.p.：小鼠中未检出；%：总占比（百分占比）。
补充材料1（S1 File）中的表C：实时荧光定量PCR（real-time PCR）所用引物。
补充材料1（S1 File）中的表D：连续传代及筛选获得高转移活性后，差异表达基因的染色体定位。拷贝数失衡（差异倍数≥2倍；P值相关，原文为p2fold; p20，疑似排版遗漏）。转移相关基因的分析基于早期和晚期传代细胞的合并数据集完成。n：受影响染色体的数量。优先上调基因以粗体标注。
补充材料1（S1 File）中的表E：SAOS和LM5细胞经连续体外传代后的大规模染色体畸变。SAOS和LM5细胞的共同受累区域以黄色标注。
补充材料1（S1 File）中的表F：SAOS和LM5细胞连续体外传代后，微阵列（microarray, MA）分析中差异表达基因与阵列比较基因组杂交（array comparative genomic hybridization, aCGH）分析中的拷贝数增减的相关性。
补充材料1（S1 File）中的图A：与拷贝数增减相关的差异表达基因的染色体定位。A）晚期与早期SAOS细胞中上调且与拷贝数增加相关的基因；B）晚期与早期SAOS细胞中下调且与拷贝数缺失相关的基因；C）晚期与早期LM5细胞中上调且与拷贝数增加相关的基因；D）晚期与早期LM5细胞中下调且与拷贝数缺失相关的基因。
补充材料1（S1 File）中的表G：SAOS/LM5细胞系统中，经筛选获得高转移活性后的大规模染色体畸变。最显著受累区域以黄色标注。
补充材料1（S1 File）中的图B：实时荧光定量PCR（real-time PCR）分析。通过微阵列（microarray, MA）和实时荧光定量PCR（qPCR）分析，比较晚期与早期传代的SAOS和LM5细胞中属于"黏着斑（Focal adhesion）"的共同上调基因。
补充材料1（S1 File）中的表H：连续体外传代后，差异倍数>2倍且P<0.05的、属于"刺猬信号通路（Hedgehog signaling pathway）"和/或"Wnt信号通路（WNT signaling pathway）"的差异表达基因。比较基准为晚期与早期传代细胞。标红的基因同时在筛选获得高转移活性后出现差异表达（补充材料1中的表K）。
补充材料1（S1 File）中的表J：根据早期和晚期传代数据集的单独比较，筛选获得高转移潜能后，属于"刺猬信号通路（Hedgehog signaling pathway）"和"Wnt信号通路（WNT signaling pathway）"的差异表达基因的富集情况。Metastatic/parental：转移组/亲本组；HH："刺猬信号通路（Hedgehog signaling pathway）"；WNT："Wnt信号通路（WNT signaling pathway）"；Total：总差异表达基因数量（差异倍数>2倍且P<0.05，在亲本或转移细胞系中经连续体外传代后出现差异表达，补充材料1中的表H）。所有4组比较的平均值已计算。
补充材料1（S1 File）中的图C：京都基因与基因组百科全书通路（KEGG pathways）。（PDF格式）