建立十二色荧光原位杂交技术鉴定食管癌KYSE450细胞系的核型

Identification of complex chromosome abnormalities in esophageal carcinoma cells KYSE450 by multicolor fluorescence in situ hybridization

目的:应用12色荧光原位杂交技术(multicolor fluorescence in situ hybridization,M-FISH)鉴定食管癌细胞系KYSE450的染色体畸变．方法:采用DOP-PCR(degenerate oligonucleotide primer-polymerase chain reaction)法混合标记两组多色painting探针,先后两次杂交到相同的KYSE450染色体片,结合反相DAPI显带技术进行核型分析．结果:成功地建立了一种重复的12色荧光原位杂交技术,并鉴定出KYSE450中存在的畸变染色体．该细胞系共有54条染色体,只有13, 21和X号染色体是正常的,其余21条染色体均表现异常．1,2,3,5,6,8,9,14,15,16和17号染色体部分区带增益,4和18号染色体部分区带缺失．7,11,12和19号染色体同时存在区带增益和其它部分区带缺失．1,2,3,4,5,6,7,8,9, 11,12,14,15,16,17,18和19号染色体显示有易位．22号染色体丢失1条,未检测到10,20,Y 染色体成分．结论:12色荧光原位杂交可用于鉴定肿瘤中复杂的染色体异常．KYSE450存在较多与原发性食管鳞癌一致的染色体改变．

AIM： To establish the technique of multicolor fluorescence in situ hybridization （M-FISH） for identification of chromosome aberrations in esophageal carcinoma cell line KYSE450. METHODS： Two pools of 12-color whole-chromosome painting （WCP） probes were designed and labeled by degenerate oligonucleotide primer-polymerase chain reaction （DOP-PCR）. FISH was performed twice on the same metaphase spreads. The karyotype was analyzed by the combination of inverted DAPI banding and M-FISH. RESULTS： Repetitive 12-color M-FISH was successfully established and the cytogenetic abnormalities in KYSE450 cells were characterized. There were 54 chromosomes in the cell line, but only those numbered 13, 21 and X were normal. DNA losses were observed at parts of chromosomes 4, 7, 11, 12, 18 and 19. Chromosomal gains and translocations occurred at chromosome 1, 2, 3, 4, 5, 6, 7, 8, 9, 11, 12, 14, 15, 16, 17, 18 and 19. Chromosome 22 showed monosomy, and no chromosomes 10, 20 and Y were detected. CONCLUSION： The established 12-color M-FISH is useful for the analysis of chromosomes in the whole genome of human tumors. KYSE450 cell line presents multiple cytogenetic abnormalities, which are in accordance with those occurred in primary esophageal squamous cell carcinoma.