荧光原位杂交和基因组半定量法检测乳腺癌组织中c-erbB-2基因扩增

Fluorescence in Situ Hybridization and Genome Semi-Quantification for Detecting Amplification of C-ERBB-2 in Breast Cancer

目的利用荧光原位杂交(FISH)与基因组半定量法检测乳腺正常良性和恶性组织中原癌基因c-erbB-2的扩增情况。方法收集50例临床切除的乳腺肿瘤组织,抽提收集样品基因组DNA并扩增c-erbB-2的特异序列构建转化子,以c-erbB-2的特异序列为探针进行间期核FISH检测乳腺肿瘤组织中c-erbB-2基因的扩增,并采用基因组半定量法比较不同组别乳腺组织中c-erbB-2基因的扩增。结果FISH检测发现正常乳腺组织中未出现c-erbB-2扩增,而良性与恶性组织中有不同程度的扩增,基因组半定量分析显示恶性肿瘤组织中c-erbB-2扩增程度高于良性与正常组织。结论c-erbB-2扩增是乳腺癌变的可靠的分子指标。可用荧光原位杂交与基因组半定量法对乳腺癌进行分子生物学诊断,且前者的敏感性与稳定性优于后者。

Objective To investigate the amplification of c-erbB-2 in breast tissues by using fluorescence in situ hybridization （FISH） and genome semi-quantification analyses. Methods 50 cases of breast tissues were collected. Genomic DNA was extracted from the collected tissues. The specific sequence of c-erbB-2 was amplified by PCR. The production of PCR was used to construct the transformers which was a probe of FISH in detecting amplification of c-erbB-2 in the interphase nucleus, and also genome semi-quantification was used to compare the amplification of c-erbB-2 in different breast tissue. Results Excessive amplification of c-erbB-2 was found in breast benign and malign tissue, but not found in normal breast tissue by FISH; The amplification level of C-erbB-2 was significantly higher in breast malign tissue than in normal and benign breast tissue. Conclusions The results suggest that the excessive amplification of C-erbB-2 might be correlated with the carcinogenesis and aggressiveness of breast cancer. Combination of the FISH and genome semi-quantification can be applied to detect the amplification of C-erbB-2, with the former more useful than the latter in terms of their sensitiveness and stability.