应用FISH和IHC技术检测中国乳腺癌患者HER-2基因状态及蛋白表达的前瞻性多中心研究

目的 对比研究乳腺癌患者IHC检测c-erbB2蛋白表达和FISH检测HER-2/neu基因扩增情况,并探讨HER-2基因状态与各临床病理特征的相关性.方法 本研究为全国73家中心参与的前瞻性研究.收集2007年10月至2009年9月乳腺癌患者标本3 249份,应用IHC和FISH两种方法分别检测3 249份乳腺癌患者手术的石蜡标本c-erbB2蛋白表达和HER-2基因扩增情况,并分析HER-2基因状态与患者各临床病理特征的关系.结果 全组患者IHC检测c-erbB2蛋白表达阳性率为46.9%(1477/3149),FISH检测HER-2基因扩增率为42.6%(1342/3149),其中IHC评分为3+和0分时,与FISH检测的一致性较高,分别为94.1%(892/948)和89.9%(660/734),而IHC 1+和2+组与FISH的一致性较低,分别为71.0%(514/725)和55.9%(415/742).同时,HER-2基因扩增与激素状态中雌激素与孕激素均阴性(r=0.45,P<0.01)、组织分级Ⅲ级(r=0.51,P<0.01)、淋巴结转移数目多于4枚(r=0.35,P<0.01)、临床分期Ⅲ/Ⅳ期(r=0.33,P<0.01)、肿瘤直径>2 cm(r=0.38,P<0.01)、绝经后(r=0.24,P<0.01)有相关性,与年龄(r=0.36,P=0.068)、CA125(r=0.11,P=0.722)、CA153(r=0.23,P=0.45)和CEA(r=0.22,P=0.074)表达情况、淋巴结转移(r=0.15,P=0.18)、肿瘤个数(r=0.21,P=0.056)及脉管瘤栓(r=0.12,P=0.133)无相关性.结论 FISH和IHC两种方法检测HER-2表达状态具有较高的一致性.结合实际情况包括费用仪器等限制,在IHC作为初筛的基础上,仍然推荐FISH作为检测HER-2基因扩增的标准方法.FISH技术检测乳腺癌患者HER-2基因表达状态可为临床指导用药和预后评价提供更可靠的依据.

A prospective multicenter study of HER-2/neu status in human breast cancer patients of mainland China: comparison of fluorescence in situ hybridization and immunohistochemistry

Objective To investigate the expression of the HER-2 in human breast carcinomas by comparing the concordance between IHC staining and FISH, and analyze the relationship between the gene expression of HER-2/neu and clinical characteristics of patients. Methods A prospective clinical trial was performed involving a large multicenter patients' samples. Totally 3 249 breast cancer samples were collected from October 2007 to September 2009 in 73 hospitals across mainland China. HER-2 status was assessed by both IHC and FISH using formalin-fixed paraffin sections of consecutive tumor samples. The relationship between the expression of HER-2/neu gene and clinical parameters was analyzed using statistical methods. Results HER-2/neu was detected by FISH in 42.6% (1342/3149) of cases, whereas IHC analysis found 46. 9% (1477/3149) of cases to be HER-2/neu positive (2 +/3 + ). A higher concordance was observed in 94. 1% (892/948) of the patients with scores of 3 + and 89. 9% (660/734) with scores of 0 + by IHC, but relatively low concordance was also observed in 71. 0% (514/725) of patients with scores of 1 + and 55. 9% (415/742) in scores of 2 + by IHC. HER-2/neu gene amplification was associated with negativity of estrogen receptor and progesterone receptor (r = 0. 45, P < 0.01) , high histological grade ( r = 0. 51,P<0.01), more than 4 positive lymph nodes (r =0. 35, P<0.01), advanced stage (r=0. 33, P< 0. 01) , large tumor size ( > 2 cm, r = 0. 38, P < 0. 01 ) , postmenopause (r = 0. 24, P < 0. 01). No statistically significant relationship was found between HER-2 gene status and the other variables including age(r=0.36, P = 0.068), CA125 (r=0.11, P=0.722) or CA153 (r = 0. 23, P=0.45) protein status, lymph node involvement (r=0. 15, P =0. 18), CEA (r = 0.22, P=0.074) , number stage of tumor (r = 0. 21, P = 0.056 ) and blood vessel invasion (r = 0. 12, P = 0. 133 ). Conclusions The comparison of IHC and FISH demonstrated an excellent correlation of HER-2/neu overexpression and gene amplification. The results availabledemonstrate that FISH-analysis as a gold standard should be performed to guide reasonable clinical treatment.