Tsuda H, Sakamaki C, Shimamura K, Hirohashi S
Pathology Division, National Cancer Center Research Institute and Hospital, Tokyo, Japan.
Acta Cytol. 1996 Jul-Aug;40(4):625-30. doi: 10.1159/000333849.
To clarify whether analysis of loss of heterozygosity (LOH) on chromosome 16q is possible using DNA isolated from fine needle aspiration specimens, a simulation study was performed using resected biopsy or mastectomy specimens of 37 breast carcinomas and 3 fibroadenomas.
A highly polymorphic (AC)n repeat region on the D16S305 locus on chromosome 16q24 was amplified in the DNA samples by the polymerase chain reaction (PCR) using 32P-labeled oligonucleotide primers, and the PCR products were electrophoresed in denaturing gel for detection of LOH by autoradiography.
PCR was successful in 34 cases, and LOH was detected in 10 (71%) of 14 carcinomas but not in the 3 fibroadenomas. These results were almost always compatible with the data obtained by restriction fragment length polymorphism analysis on chromosome 16q using Southern blotting.
Examination of LOH on 16q by (AC)n polymorphism analysis using fine needle aspiration specimens is suggested as a supportive tool for preoperative diagnosis of breast tumors.
为了阐明使用细针穿刺标本中分离的DNA是否能够分析16号染色体q臂上的杂合性缺失(LOH),我们使用37例乳腺癌和3例纤维腺瘤的切除活检或乳房切除术标本进行了一项模拟研究。
使用32P标记的寡核苷酸引物,通过聚合酶链反应(PCR)在DNA样本中扩增16号染色体q24上D16S305位点的一个高度多态性(AC)n重复区域,并将PCR产物在变性凝胶中进行电泳,通过放射自显影检测LOH。
34例PCR成功,在14例癌中的10例(71%)检测到LOH,而3例纤维腺瘤中未检测到。这些结果几乎总是与使用Southern印迹法对16号染色体q臂进行限制性片段长度多态性分析所获得的数据一致。
建议使用细针穿刺标本通过(AC)n多态性分析检测16q上的LOH,作为乳腺肿瘤术前诊断的一种辅助工具。