Application Of Fluorescence In Situ Hybridization And Study On Cyclooxygenase-2 In Esophageal Squamous Cell Carcinoma

Esophageal Squamous Cell Carcinoma (ESCC) is one type of malignant tumors derived from the esophageal epithelial cells with high incidence in North China. Despite surgical treatment and chemotherapy, the prognosis of ESCC is still poor. The development of ESCC is a multifactor and multistage process involving quite some genes abnormality.Chromosome aberration is one of the distinctive features of tumors, which many cytogenetic and molecular technologies are developed to analyze. Fluorescence in situ hybridization (FISH) is one of the most used techniques because of its high sensitivity, specificity and ratio of signal to noise and both application on structural and numerary aberration of chromosomes.In the present study, application of FISH was attempted according to level of chromosome, chromosome region and gene. Repetitive multicolour FISH (RM-FISH) was performed on cell line KYSE140 in search of chromosome aberrations. The 24 chromosome paintings (twenty-two pairs of autosomes plus the two sex chromosomes) were tagged with different combination of Cy3, Cy5, FITC and DEAC, thus two pools of twelve color painting probes were constructed and were performed on the same metaphase in sequence respectively. In order to detect changes of regions on the same chromosome occurring in no synchronization, three regions of 3q, 8q and 11q were selected respectively and labeled with Cy3, Cy5 or FITC, to be performed on the cell line EC9706. To detect gene amplification rapidly at low expense, a new slide preparation was established. One single drop of each nuclei suspension from sixteen tumors, in much less volume than that of the standard, was dropped on the slide, which formed an array on the slide. The amplification of Nek2 and Hec1 gene was detected in clinical tissues of ESCC in use of gene-specific BAC (bacterial artificial chromosome) probe labeled with Cy3 by the nuclei array FISH which we named.The results of RM-FISH showed that aberration took place in most chromosomes of KYSE140. The number of chromosomes increased obviously in chromosome 1, 2, 3, 7 and...