应用快速引物原位标记技术检测鼻咽癌染色体异常

摘要

Objective The chromosomal aberration is a common event in nasopharyngeal carcinoma (NPC). The currently-used methods to detect chromosomal abnormalities are complicated and of limited clinical value. The purpose of the present study is to explore the feasibility of detecting chromosomal abnormalities in NPC by rapid primed in situ labeling (PRINS). Methods The copy changes of Chromosome 3 and 7 in the frozen section of 15 cases of NPC and five cases of normal nasopharyngeal mucosa were detected using rapid PRINS and oligonucleotide primers specific for chromosome 3 and 7. The standards of the chromosomes numerical aberration are as the following: the ratio of cells with one or no labeling signals at 65% or more was defined as the losses of chromosome; the ratio of cells with three or more labeling signals at 6.5% or more was defined as the gain of chromosome copy numbers. Results Of 15 cases of carcinoma tissues, the labeling rate of chromosome 3 and 7 was 87.4% and 88.6% respectively. The gain of chromosome 3 copy numbers was observed in ten cases ( 66.7%) and the loss of chromosome 7 was detected in five cases ( 33.3%). Four cases of them had both the gain of chromosome 3 copy numbers and the loss of chromosome 7. In normal nasopharyngeal tissues the labeling rate of chromosome 3 and 7 was 92% and 91.8% respectively, and diploid cells were 43.2% and 43.6% respectively, without trisomy. The difference of diploid cells between primary NPC and normal nasopharyngeal mucosa was statistically significant (P< 0.05). Conclusion The results indicate that rapid PRINS method can be used to detect interphase chromosomes in frozen tissue sections and chromosomes numerical aberration, which may be helpful to the diagnosis of NPC.%目的探讨快速引物原位标记技术检测鼻咽癌染色体异常的可行性.方法采用快速引物原位标记技术,分别以人3号和7号染色体特异性寡核苷酸作引物,检测15例鼻咽癌和5例鼻咽正常冰冻组织切片细胞染色体,染色体异常标准以标记信号≤1的细胞比例≥65%时视为染色体丢失,标记信号≥3的细胞比例≥6.5%作为染色体拷贝数增加.结果鼻咽癌组织细胞3号染色体标记率为 88.6%,10例(66.7%)染色体拷贝数增加;7号染色体标记率 87.4%,5例(33.3%)染色体丢失;其中4例同时存在3号染色体拷贝数增加和7号染色体丢失.正常组织3号和7号染色体标记率分别为92%和 91.8%,二倍体细胞分别为 43.2%和 43.6%,与鼻咽癌比较差异均有统计学意义(P< 0.05),未发现三体细胞.结论快速引物原位标记技术可用于鼻咽癌冰冻组织切片中染色体的检测,染色体数目的改变可能有助于鼻咽癌的诊断.