Singh B, Gogineni S K, Sacks P G, Shaha A R, Shah J P, Stoffel A, Rao P H
Laboratory of Epithelial Cancer Biology, Head and Neck Service, Memorial Sloan-Kettering Cancer Center, New York, NY 10021, USA.
Cancer Res. 2001 Jun 1;61(11):4506-13.
We applied a combination of molecular cytogenetic methods, including comparative genomic hybridization (CGH), spectral karyotyping (SKY), and fluorescence in situ hybridization, to characterize the genetic aberrations in a panel of 11 cell lines derived from head and neck squamous cell carcinoma and 1 cell line derived from premalignant oral epithelium. CGH identified recurrent chromosomal losses at 1p, 3p, 4, 8p, 10p, and 18q; gains at 3q, 5p, 8q, 9q, and 14q; and high-level amplification at 3q13, 3q25-q26, 5q22-q23, 7q21, 8q24, 11q13-q14, 12p13, 14q24, and 20q13.1. Several recurrent translocations including t(1;13)(q10;q10), t(13;13)(q10;q10), t(14;14)(q10;q10), i(8)(q10), and i(9)(q10) and breakpoint clusters at 1p11, 1q21, 3p11, 5q11, 5q13, 6q23, 8p11, 8q11, 9p13, 9q13, 10q11, 11q13, 13q10, 14q10, and 15q10 were identified by SKY. There was a good correlation between the number of aberrations identified by CGH and SKY (r = 0.69), and the analyses were both confirmatory and complementary in their assessment of genetic aberrations. Amplification at 3q26-q27 was identified in 42% of cases. Although SKY defined the derivation of 3q gain, the precise breakpoint remained unassigned. Positional cloning efforts directed at the amplified region at 3q26-q27 identified three highly overlapping nonchimeric yeast artificial chromosome clones containing the apex of amplification. The use of these yeast artificial chromosome clones as a probe for fluorescence in situ hybridization analysis allowed a detailed characterization and quantification of the 3q amplification and refinement of unassigned SKY breakpoints.
我们应用了多种分子细胞遗传学方法的组合，包括比较基因组杂交（CGH）、光谱核型分析（SKY）和荧光原位杂交，以表征一组源自头颈部鳞状细胞癌的11个细胞系和1个源自癌前口腔上皮的细胞系中的基因畸变。CGH鉴定出1p、3p、4、8p、10p和18q存在反复的染色体缺失；3q、5p、8q、9q和14q存在染色体增加；以及3q13、3q25 - q26、5q22 - q23、7q21、8q24、11q13 - q14、12p13、14q24和20q13.1存在高水平扩增。SKY鉴定出几种反复出现的易位，包括t(1;13)(q10;q10)、t(13;13)(q10;q10)、t(14;14)(q10;q10)、i(8)(q10)和i(9)(q10)，以及在1p11、1q21、3p11、5q11、5q13、6q23、8p11、8q11、9p13、9q13、10q11、11q13、13q10、14q10和15q10处的断点簇。CGH和SKY鉴定出的畸变数量之间存在良好的相关性（r = 0.69），并且这两种分析在评估基因畸变方面既具有确证性又具有互补性。在42%的病例中鉴定出3q26 - q27处的扩增。虽然SKY确定了3q增加的来源，但精确的断点仍未确定。针对3q26 - q27扩增区域的定位克隆工作鉴定出三个高度重叠且非嵌合的酵母人工染色体克隆，其包含扩增顶点。使用这些酵母人工染色体克隆作为荧光原位杂交分析的探针，能够对3q扩增进行详细的表征和定量，并细化未确定的SKY断点。
