双探针显色和荧光原位杂交法检测乳腺癌HER2基因状态和17号染色体的分析
中华病理学杂志, 2010, 39(3): 161-165.
摘 要：
目的 通过与荧光原位杂交(FISH)比较,评估双探针显色原位杂交(双探针CISH)法在诊断女性乳腺浸润性导管癌HER2基因状态中应用的可靠性,同时探讨17号染色体多倍体对原位杂交诊断HER2基因状态时可能产生的影响.方法收集146例乳腺癌组织的常规石蜡包埋标本,分别用欧盟(CE)认证的FISH(146例)及CISH(73例)HER2/17号染色体着丝粒(CENl7)双探针试剂盒技术,对肿瘤标本的HER2基因状态进行检测,并按照2007年美国临床肿瘤协会及美国病理家协会(ASCO/CAP)标准分别用计算HER2基因拷贝数和(或)计算HER2/CENl7比值的方法对结果进行分析.结果在同时进行FISH和双探针CISH检测的73例标本中发现,两种技术对HER2阴性和阳性的诊断符合率分别为91.7%(33/36)和97.4%(37/38),两者的总体符合率为95.9%(70/73);在对146例FISH结果的计数分析中,计数HER2基因拷贝数得出不确定诊断的病例量是计数HER2/CENl7得出不确定诊断病例量的1.6倍(13/8);此外,在FISH和CISH结果中按拷贝数计数为阳性的病例与在按比值计数时却为阴性病例的不吻合率分别为4.8%(3/63)和3.O%(1/33);同时在FISH HER2阳性病例(HER2/CENl7)中多倍体发生率为63.5%(40/63,P=0.002),高于阴性者中多倍体的发生率(37.3%,28/75).结论 双探针CISH技术检测乳腺癌HER2基因状态的结果和FISH技术检测的结果非常一致,提示两种技术临床诊断价值相近,并且同步检测并计数HER2和17号染色体有助于明确HER2基因状态.
双探针显色原位杂交技术检测HER2基因的应用
临床与实验病理学杂志, 2010 (005): 627-629.
摘 要：HER2是调节细胞生长过程中起关键作用的跨膜蛋白，编码基因定位于17q21。乳腺癌患者中，约20％～30％存在HER2基因或蛋白的过表达，导致肿瘤细胞过度增殖，使肿瘤的恶性程度远高于HER2未过度表达的患者。近年来，HER2的状态被公认为是判断乳腺癌预后风险和决策靶向治疗的重要指标。
Comparison of dual-probe CISH with FISH in the determination of HER2/chromosome 17 for patients with creast cancer in China
J Clin Oncol (Meeting Abstracts). 2010, 28(15_suppl): 1149.
The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China; GeneDiagnostics, Inc., Hangzhou, China
Abstract
Background: The aim of this investigation was to evaluate the clinical application of dual-probe chromogenic in situ hybridization (CISH) in the testing of HER2 gene status in patients with breast cancer by comparison with fluorescence in situ hybridization (FISH). The potential impact of chromosome 17 polysomy in the determination of HER2 status was also evaluated.
Methods: We collected 146 paraffin-embedded tissue sections of breast cancer and determined HER2 gene and chromosome 17 copy numbers using CE approved commercial kits of dual-probe FISH (for 146 cases) and dual-probe CISH (for 73 cases), respectively. The results were interpreted based on either HER2 gene copy number or the ratio of HER2/chromosome 17 (ASCO/CAP, 2007).
Results: Of the 73 cases measured by both FISH and CISH methods, the concordance between two methods for negative and positive results was 91.7% and 97.4%, respectively, while the overall concordance between the two methods was 95.9%. Of the 146 cases measured by FISH, 13 cases were interpreted as equivocal if only HER2 copies were counted, which was 1.6-fold (13/8) higher than the equivocal cases interpreted by counting the ratio of HER2/chromosome 17. Moreover, 3 of 63 (4.8%) HER2 positive cases determined by HER2 copies turned out to be HER2 negative determined by the ratio of HER2/chromosome 17, In CISH results, 1 of 33 (3.0%) positive cases was turned to negative by the comparison of two corresponding scoring methods. We also observed from FISH results that there were more chromosome 17 polysomy cases (63.5%, 40/63) in HER2 positive subgroup than those (37.3%, 28/75) in HER2 negative subgroup. (p=0.002). In addition, CISH method, which simultaneously shows HER2 signals and morphology, has obvious advantages in HER2 determination of one case with DCIS.
Conclusions: Our HER2 results determined by dual-probe CISH method were almost perfect agreed with the corresponding results determined by FISH method, indicating dual-probe CISH is a reliable alternative to FISH in HER2 testing. This study also indicates that the diagnostic accuracy may be increased when HER2 and chromosome 17 are simultaneously tested and counted.
