염색체 미세절단법을 이용한 표지염색체의 전좌 부위에 대한 DNA Painting Probe 제작

Abstract

Purpose: Chromosome microdissection has been recommended as a technology to overcome the limited problems of conventional cytogenetic analysis and is a direct approach to isolate DNA from specific interesting region of chromosome. KUMA-1 cell line has a specific reserved chromosome abnormality during process from primary cancer culture to continuous cell line development, der(2)t(2;?)(qter;?). So molecular analysis for translocation region of der(2) may be helpful to understand pathogenesis of this primary cancer. The aim of this study was to develop painting probe for the translocation region for molecular study in future about translocation region of der(2) of KUMA-1 cell line. Materials and Methods: KUMA-1 cell line was derived from a squamous cell carcinoma of urinary bladder. The translocation breakpoint region of der(2) appeared in KUMA-1 cell line was microdissected and dissected chromosome segments were amplified by PCR reaction. Fluorescent in situ hybridization was conducted on KUMA-1 metaphase cells with the probe generated from PCR product to confirm the construction of painting probe containing the translocation breakpoint of der(2). Results: Painting probe was hybridized to the metaphase chromosome of KUMA-1 cell line and two fluorescent signals were mapped to the translocation forming chromosomal region of der(2). Conclusion: It was possible to construct the painting probe for the translocation region of der (2) by chromosome microdissection.