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

A prospective multicenter study of HER-2/neu status in human breast cancer patients of China's Mainland. comparison of fluorescence in situ hybridization and immunohistochemistry

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.

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 China's Mainland. 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.