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  • br Amplification of the genes located on

    2022-05-13


    Amplification of the genes located on 8p and 8q is summa-rized in Fig. 2A. Amplifications of FGFR1 and ZNF703 were observed at similar frequencies. By dual-color FISH, the sig-nals for ZNF703, FGFR1, ADAM5, and IKBKB, if amplified, were found to be closely associated, suggesting location on the same amplicons. The observed amplicons were of various types, including HSR (Fig. 1A), CoPoly (Fig. 1C), LA (Fig. 1G), and Mono (Fig. 1I). However, except for eight cases of HSR type, all of the amplicons exhibited the PC subtype (Fig. 2A), and thus were considered to be entopically located. In Case 8 (Fig. 1C-F), the tumor nuclei had large signals for Bleomycin Sulfate 8 surrounded by numerous amplified gene signals for 8p (Fig. 1D, large arrows); this specimen also yielded small paired signals for the 8p genes and centromere 8 (Fig. 1D, small arrows).
    Although MYC also was amplified in a variety of amplicon types, DM (Fig. 1J) and CoPoly types were observed the most frequently (Fig. 2A). The PC subtype was detected only in two LA-type and one CoPoly amplicons. In Case 53, tight or loosely clustered signals for MYC and centromere 8 were ob-served (Fig. 1K and L). Co-localization of MYC and MTDH, and of MYC and PRDM14, in single nuclei were found in ten and four cases, respectively. Co-amplification of the genes located on 8p and 8q was observed in seven tumors; however, in each case the two amplicons were separated.
    The amplification of CCND1 was found in 40 tumors; 31 of these tumors exhibited the HSR type and 21 of the HSR-type amplicons were of the PC subtype (Fig. 2B). In Cases 69 and 76, increased numbers of CCND1 signals associated with small signals for centromere 11 were observed, as shown
    Amplicons in breast cancers 37
    Fig. 1 Amplifications of FGFR1 (Cases 30, 8, and 9) (A-H), ZNF703 (Case 18) (I), and MYC (Cases 19 and 53) (J-L) (orange, gene-specific signals; green, signals for the centromeric regions on which the specific genes are located). In Panel D, paired signals of FGFR1 and centromere 8 (small arrows) and large signals of centromere 8 surrounded by numerous FGFR1 (large arrows) signals were observed (D, triple-band filter; E, SpectrumGreen™-spe-cific filter; F, SpectrumOrange™-specific filter). Panels K and L showed co-amplification of MYC and centromere 8. PC: PC-subtype. Scale bar: 10 μm.
    Amplicons in breast cancers 39
    Fig. 3 Amplicon on 11q13. Overlapped loose clusters of CCND1 (orange) and small signals of centromere 11 (green) were found in the imprinted cells (A, triple-band filter; B, SpectrumGreen™-specific filter; Case 69). Dual-color FISH on FFPE tissue shows co-localization of CCND1 (green signals) and C11ORF30 (orange signals) (C, Case 4). Scale bar: 10 μm.
    in Fig. 3A and B. Among tumors that yielded amplification of CCND1, 19 of 40 (48%) exhibited co-amplification of C11ORF30 (Fig. 3C). Conversely, no amplification of C11ORF30 was observed without co-amplification of CCND1.
    The HSR-type amplification of ERBB2 was found in 31 tu-mors (Fig. 2C). In eight of these 31, the clustered ERBB2 sig-nals were found near CEP17 signals (PC subtype); in the other
    Fig. 4 Amplicon on 17q. CoPoly-type, PC-subtype amplification of ERBB2 (orange) and centromere 17 (green) on imprinted cells (A, Case 103; B, Case 29). Numerous minute paired signals were found in (A); small ERBB2 signals aggregated around the large centromeric signals were found in (B). Normal lymphocytes with two paired signals are seen at the corners. Note the co-amplification of CPD (orange) and ERBB2 (green) in Case 83 (C), and of BIR5 (orange) and ERBB2 (green) in Case 14 (D, triple-band filter: E, SpectrumGreen™-specific filter). Scale bar: 10 μm.
    Table 2 Co-amplification of non-syntenic genes
    Case No.
    H L
    H H H
    H M
    H
    H H
    H H H
    M
    H L
    H