【摘要】 目的端粒为染色体末端的非编码重复序列,对维持染色体的稳定及细胞复制和死亡具有重要的作用。细胞端粒重复序列的丢失与衰老和肿瘤的发生相关,在人体多种肿瘤的研究中,人们发现端粒长度的测定对诊断及预后判断具有重要价值。为了寻求一种端粒精确定量的新方法,开展了本实验研究。方法应用肽核苷酸探针(PNA)标记荧光原位杂交(FISH)—荧光素异硫氰酸(FITC)法检测黑色素瘤、转移黑色素瘤及黑色素痣石蜡切片中的端粒,以荧光显微镜、激光共聚焦显微镜生成图像,并详细记录细胞核平均面积、端粒数目、端粒面积。通过对采集图像的分析,对端粒进行了精确定量。结论FISH/FITC提供了一种方便、快速检测、精确定量组织切片中端粒的方法;定量FISH能在组织/细胞水平上提供端粒更特异的信息。更多还原

【Abstract】 Objective: Telomeres containing noncoding DNA repeats at the end of the chromosomes are essential for chromosomal stability and are complicated in regulating the replication and senescence of cells.The gradual loss of telomere repeats in cells has been linked to aging and tumor development.In some human cancers telomere length measurement has proved to be of diagnostic and prognostic value.This study was carried out to find a quantification method for telomere.Methods PNA(Peptide nucleic acid probe) conjugated FISH(Fluorescence in situ hybridization)-FITC(Fluorescein isothiocyante) was performed on paraffin sections from melanoma,melanoma metastases and common naevi,respectively.Telomere mean maximum diameter,area and number per nuclear area were calculated by image analysis on fluorescence images elaborated through Confocal Laser Scanning Microscope(CLSM) and fluorescence microscope.Conclusions The telomere FISH/FITC provides a convenient,rapid method for detection of the telomeric sequences in nuclei in samples from paraffin tissues.Telomere Quantitative FISH could give more specific informations at the cellular level in telomere investigation.更多还原