不同pH值的40mL/L磷酸盐缓冲甲醛液固定乳腺癌组织对FISH法检测HER2基因结果的影响  

Comparison of HER2 gene amplification detected by FISH assay for breast cancer tissues fixed by different pH values of 40mL/L phosphate buffered formalin

作　　者：陈志强 米贤军 陈昂 段立锋 刘超凡 邓文同 CHEN Zhi-qiang;MI Xian-jun;CHEN Ang;DUAN Li-feng;LIU Chao-fan;DENG Wen-tong(Department of Pathology,Zhongshan Bo ai Hospital Affiliated to Southern Medical University,Zhongshan 528400,China)

机构地区：[1]南方医科大学附属中山博爱医院病理科,广东中山528400

出　　处：《西安交通大学学报（医学版）》2018年第3期438-441,共4页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：广东省医学科学技术研究基金立项课题(No.A2017321);广东省中山市卫生和计划生育局医学科研立项课题(No.2017J111;2014J128)~~

摘　　要：目的探讨不同pH值的40mL/L磷酸盐缓冲甲醛固定液对荧光原位杂交(fluorescence in situ hybridization,FISH)法检测乳腺原发性浸润癌人类表皮生长因子受体2(human pidermal growth factor receptor 2,HER2)基因扩增结果的影响。方法选取中山市博爱医院2014年3月至2017年1月期间,手术切除的乳腺癌75例,同一肿瘤病变部位取材4块,按照固定液pH不同,随机分为A、B、C、D组。固定液使用40mL/L磷酸盐缓冲甲醛,A^D组固定液pH值分别为6.0、7.0、7.5、8.0,分别固定15h后制作石蜡切片。采用FISH法检测HER2基因扩增情况,比较各组HER2基因扩增检测结果的差异。结果 (1)在荧光显微镜下,A、D组的组织轮廓较模糊、出现细胞碎片、部分探针定位欠佳、出现信号不规则甚至丢失、核溶解现象,见明亮的桔红/绿荧光信号。B、C组的组织轮廓清晰而完整、背景洁净、探针定位准确,见耀眼的桔红/绿荧光信号。(2)B、C组HER2基因扩增阳性率稳定在24.00%~25.33%;B、C组的基因扩增阳性率显著高于A、D组(P<0.05)。结论采用荧光原位杂交法检测乳腺原发性浸润癌HER2基因扩增,固定液的pH值在7.0~7.5之间可获得可靠的检测结果。Objective To investigate the effects of 40 mL/L phosphate buffered formalin fixative solution with different pH values on the amplification result of human epidermal growth factor receptor 2(HER2)gene in breast cancer by fluorescence in situ hybridization(FISH).Methods We collected 75 cases of primary invasive breast cancer submitted by the Gynecology Department of Zhongshan Bo ai Hospital from March 2014 to January 2017.Four samples were taken from the same tumor lesion site.According to pH value of the fixatives,we divided them into A,B,C,and D groups.Phosphate buffered formaldehyde of 40 mL/L was used as the fixative.The pH value for Group A to Group D was 6.0,7.0,7.5,and 8.0,respectively.Paraffin sections were made after 15 hours of fixation separately.The amplification of HER2 gene was detected by FISH.Differences in HER2 gene amplification detection results were compared.Results ①Under the fluorescence microscope,the tissue contour in Groups A and D was vague.Cell debris appeared and some of the probe was positioned poorly.Besides,signal irregularity and even loss and nuclear emptying occurred;bright orange or green fluorescent signals could be seen.However,the tissue contour was clear and complete in Groups B and C.The background was clean.The probe was positioned accurately.Dazzling orange or green fluorescent signals could be seen.②The positive rate of HER2 gene amplification was stable at 24.00%-25.33%.So it turned out that the positive rate of gene amplification was significantly higher in Groups B and C than in Groups A and D(P<0.05).Conclusion When FISH is used to detect HER2 gene amplification in primary breast infiltrating carcinoma,a pH value from 7.0 to 7.5 can obtain trust worthy detection results.

关 键 词：乳腺原发性浸润癌 pH值 荧光原位杂交(FISH) 人类表皮生长因子受体2(HER2) 

分 类 号：R36[医药卫生—病理学]