Rapid Detection of the Epidermal Growth Factor Receptor Mutation in Non-Small-Cell Lung Cancer for Analysis of Acquired Resistance Using Molecular Beacons

A secondary mutation (T790M) in epidermal growth factor receptor (EGFR) is a hallmark of acquired resistance to EGFR inhibitors used to treat non-small-cell lung cancer (NSCLC). Therefore, identifying the T790M mutation is crucial to guide treatment decisions. Given that DNA sequencing methods are time-consuming and insensitive, we developed and investigated the feasibility of using molecular beacons for the detection of the T790M mutation in EGFR. A molecular beacon complementary to the region of the secondary EGFR mutation (T790M) was designed and used in NSCLC samples bearing drug-sensitive and -resistant EGFR mutations. For a rapid and simple assay, we attempted to use the molecular beacon with real-time PCR and in situ fluorescence imaging. The ability of the designed molecular beacon to specifically detect the T790M mutation of EGFR was tested for samples from two patients with drug resistance and compared with conventional DNA sequencing methods. The molecular beacon successfully detected the T790M mutation in patient samples with drug resistance. The sensitivity of the molecular beacon, which detected as little as 2% of genomic DNA from the drug-resistant cells (H1975), was much higher than direct sequencing. Furthermore, in situ fluorescence imaging with the molecular beacon gave rise to a distinguishable signal for the T790M mutation in drug-resistant cells. The molecular beacon-based approach enabled rapid and sensitive detection of the EGFR mutation (T790M) in NSCLC with in situ fluorescence imaging, which can be directed to determine various treatment options in patients with cancer.Lung cancer is the leading cause of cancer-related death and is responsible for 1.3 million deaths worldwide annually. Despite marked progress in cancer treatment, the 5-year survival rate of patients with lung cancer has remained at around 15% for over the past 30 years.¹ In an attempt to provide improved treatment, a variety of targeted therapeutics have been investigated based on the progress in our understanding of the molecular mechanisms underlying lung tumorigenesis. The epidermal growth factor receptor (EGFR) is one such therapeutic target in non-small-cell lung cancer (NSCLC), which has led to the development of effective anticancer agents, such as EGFR tyrosine kinase inhibitors (EGFR-TKIs). EGFR-TKIs, including gefitinib and erlotinib, have been approved for clinical treatment and are currently prescribed in patients with advanced NSCLC. Extensive clinical trials of EGFR-TKIs have shown a survival benefit associated with EGFR-TKIs in patients with lung cancer.² Clinical and molecular analyses have revealed that EGFR mutations in the kinase domain, most commonly small in frame deletions in exon 19 or a substitution mutation of leucine 858 to arginine (L858R) in exon 21, are associated with a high clinical response rate of tumors to gefitinib or erlotinib in patients with NSCLC.^3,4Most patients carrying EGFR mutations initially show an impressive response to EGFR-TKIs. However, they eventually develop resistance after prolonged treatment. Previous studies have revealed that a secondary EGFR mutation, as a result of a threonin-to-methionine amino acid change at the 790th codon (T790M), is often observed in the tumors of patients with NSCLC that develop acquired resistance.^4,5 From the clinical standpoint, detection of the secondary mutation (T790M) in patients who have disease progression after prolonged treatment with gefitinib or erlotinib is crucial to planning alternative treatment options.Various methods for the detection of EGFR mutations in patient samples have been developed.^6,7,8 Currently, the most common method for the detection of EGFR mutations in tumor tissue samples is direct sequencing of EGFR fragments amplified from genomic DNA. Since the amount of sample available for clinical analysis from patients with advanced NSCLC is limited, and often mixed with an excess amount of normal tissue, the detection of a mutation by direct sequencing may be inaccurate if the number of cancer cells is below a minimum of 20% of the total specimen.⁸ Therefore, an issue of great importance is the development of methods that allow for unambiguous detection of EFGR mutation in patients with cancer in a rapid and sensitive way.A molecular beacon (MB) is a nucleotide strand in a distinct stem-loop structure with a fluorescence dye attached on one end and a quencher dye on the other.^9,10 In the hairpin conformation, the proximity between the fluorescence-quencher pair allows quenching due to fluorescence resonance energy transfer.¹¹ Only when hybridized to the specific target, the probe separates and expands its stem region, pushing away the dye-attached-ends of the complex. This results in the restoration of the fluorescence unaffected by the distant quencher. As the fluorescence intensity varies according to the changes in the probe conformation to the target DNA, a much more specific and selective signal can be detected. Applying a molecular beacon probe to real-time PCR allows for rapid, simple, and cost-effective detection of the target without post-PCR manipulation.^12,13,14 Even though the molecular beacon has been used for a variety of purposes, there has been no prior report on its application to the analysis of mutations in the EGFR gene.In an effort to develop a rapid and sensitive assay for the identification of acquired resistance to EGFR-TKIs in patients with NSCLC, we demonstrate the use of a molecular beacon for the detection of the secondary mutation, T790M, in EGFR by real-time PCR and in situ fluorescence imaging approaches. Under optimized molecular beacon and real-time PCR conditions, we determined that the detection sensitivity was comparable to or better than other recently developed EGFR mutation detection systems. Furthermore, fluorescence imaging of cells with the T790M mutation using the molecular beacon also provided a simple and rapid detection method for acquired resistance in situ. Therefore, the molecular beacon-based EGFR mutation detection system offers a potentially useful technical platform for the clinical diagnosis of lung cancer samples and ultimately might help to guide treatment decisions for lung cancer patients.     

湖北地区非小细胞肺癌EGFR 基因突变及其意义的研究

目的 探讨湖北地区非小细胞肺癌(NSCLC)患者中EGFR基因的突变情况,评价湖北地区EGFR突变的检测力度,为临床合理选择酪氨酸激酶抑制剂提供可靠的实验依据。方法 采用PCR-sanger测序法以及ADx-ARMS法检测2010～2015年期间253例肺癌标本中EGFR基因突变情况,探讨不同检测手段的敏感度,探讨不同性别、不同组织分型间EGFR不同外显子的突变频率差异及其临床意义。结果253例肺癌标本中共检测到突变位点93个,以E19和E21的突变最为常见,占比为53.76%和35.38%。ADx-ARMS法的EGFR总突变检出率明显高于Sanger测序法(P=0.001)。女性肺癌患者的检出率明显高于男性(P=0.001)。不同组织分型中,腺癌(38.01%)的检出率最高,腺鳞癌(30.77%)与大细胞癌(20%)次之,鳞癌(4.55%)较低。无吸烟史的肺癌患者检出率(51.6%,81/157)明显高于有吸烟史患者(24%,12/50)。结论与PCR-sanger测序法相比,ADx-ARMS法对临床筛选EGFR-TKI治疗的受益亚群,预测EGFR-TKI疗效更为敏感有意义。湖北地区女性肺癌患者中EGFR突变频率明显高于男性,与激素水平变化及吸烟状况有关。     

成纤维细胞生长因子受体1在非小细胞肺癌中的研究进展

非小细胞肺癌(non-small cell lung cancer, NSCLC)已成为全世界癌症患者死亡的主要原因之一, 其治疗成为目前研究热点。但NSCLC中成纤维细胞生长因子受体(fibroblast growth factor receptor, FGFR)基因突变以及间变性淋巴瘤激酶(anaplastic lymphoma kinase, ALK)融合基因发生率低, 相应靶向药物治疗的总体效果并不理想。FGFR1作为NSCLC中较常见的异常基因, 其异常扩增高表达与多种肿瘤的发生发展相关, 同时发现多种相关的分子靶向药物对肿瘤有抑制作用。本文对FGFR1在NSCLC中的扩增高表达情况、其与NSCLC临床特点之间的关系以及目前相关靶向药物治疗研究进展作一综述。     

血清表皮生长因子与早产儿肺发育相关研究

目的 探讨血清表皮生长因子水平与早产儿肺发育的相关性.方法 选择住院早产儿35例.根据胎龄分成3组,分别是①28周≤胎龄〈32周,9例,②32周≤胎龄〈35周,14例,③35周≤胎龄〈37周,12例,同时根据有无并发早产儿肺透明膜病（HMD）分为HMD组19例,非HMD组16例;对照组选择足月儿高胆红素血症住院患儿12例.无其他并发症.采用酶联免疫吸附测定法（ELISA）,35例早产儿和12例对照组足月儿分别于出生1h、24h、72h、和生后第7天空腹股静脉采血测定血清表皮生长因子水平.结果 早产儿组出生1h、24h、72h、和生后第7天.血清表皮生长因子含量水平均呈递增关系,差异有显著性;早产儿组与足月儿组血清表皮生长因子含量水平于出生1h、24h、72h差异有显著性,生后第7天差异无显著性;在生后1h、24h、72h,早产儿有HMD组与无HMD组患儿血清表皮生长因子含量水平差异有显著性.结论 早产儿出生后血清表皮生长因子含量水平呈递增关系,提示早产儿出生后体内EGF合成和释放增加,可能促进肺脏器成熟;早产儿组与足月儿组血清表皮生长因子含量水平出生后差异有显著性,早产儿出生后血清EGF浓度水平和HMD的严重程度正相关,提示在新生儿早期检测血清表皮生长因子含量可以作为HMD发病的预测指标.     

非小细胞肺癌表皮生长因子受体基因及k-ras基因突变与临床病理特征的关系

  目的  探讨非小细胞肺癌(non-small cell lung cancers, NSCLC)肿瘤组织中表皮生长因子受体(epidermal growth factor receptor, EGFR)基因及k-ras基因突变与临床病理特征的关系。  方法  应用显微切割技术及蝎形探针扩增阻滞突变系统(scorpions amplification refractory mutation system, Scorpions ARMS)检测170例NSCLC石蜡包埋肿瘤组织中EGFR基因第18、19、20和21外显子突变及k-ras基因12、13密码子突变, 统计分析EGFR及k-ras基因突变与不同组织类型NSCLC临床和病理特征的相关性。  结果  Scorpions ARMS检测结果显示, 170例NSCLC肿瘤组织中84例存在EGFR突变, 突变检出率为49.4%, 其中39例为EGFR第19外显子缺失改变, 34例为EGFR第21外显子L858R点突变, 3例为EGFR第21外显子L861Q点突变, 4例为EGFR第20外显子插入突变, 2例为EGFR第20外显子S768I点突变, 另外2例为EGFR第20外显子T790M点突变和第21外显子L858R点突变同时存在。170例NSCLC肿瘤组织标本中检测到14例k-ras基因突变, 检出率为8.2%, 均位于12密码子, 其中8例为12CYS点突变, 3例为12ASP点突变, 3例为12VAL点突变。未发现k-ras基因突变与EGFR基因突变发生在同一NSCLC肿瘤组织标本中。腺癌与非腺癌EG-FR基因突变比较差异具有统计学意义(P < 0.001), 突变更常见于腺癌。此外, 比较EGFR基因突变与NSCLC各临床病理因素的关系, 发现EGFR基因突变更常见于女性、不吸烟、肿瘤较小(≤ 3cm)且分化程度较好的NSCLC患者; 然而, k-ras基因突变与患者年龄、性别、吸烟史、肿瘤大小、组织学类型、肿瘤分化程度、淋巴结转移及pTNM分期等均无显著的相关性(P > 0.05)。  结论  中国NSCLC患者的EGFR基因突变检出率明显高于其他国家, 且女性、不吸烟、腺癌患者突变率较高; 而k-ras基因突变的检出率很低, 其突变与性别、年龄等临床病理特征无关; EGFR和k-ras基因突变不会同时存在同一患者中。吉非替尼疗效和耐药与EGFR和k-ras基因突变相关。Scorpions ARMS技术可快速、敏感、准确地检测EGFR与k-ras基因突变, 能为临床治疗及预后提供重要信息。     

非小细胞肺癌患者不同类型样本中表皮生长因子受体基因突变的差异研究

目的 检测238例非小细胞肺癌(NSCLC)患者不同类型样本中表皮生长因子受体(EGFR)基因的突变情况,探讨不同类型样本在EGFR基因突变中的应用价值。方法 采用扩增阻碍突变系统(ARMS)检测238例NSCLC中EGFR基因18～21外显子的突变情况,探讨不同样本类型、不同性别、不同年龄、不同病理分型和不同临床分期的EGFR突变频率的差异及临床意义。结果 238例NSCLC患者EGFR总的突变率为44.5%(106/238),其中组织、血浆和胸腔积液样本分别为105例,115例和18例,其突变率分别为52.4%(55/105),34.8%(40/115)和61.1%(11/18),与组织样本相比,血浆样本的EGFR突变检出率明显低于组织样本,差异具有统计学意义(χ2=6.93,P<0.05),但组织与胸腔积液样本相比其检出率差异无统计学意义(χ2=0.471,P>0.05)。女性NSCLC患者的EGFR突变的检出率明显高于男性患者,差异有统计学意义(55.3% vs 36.3%,χ2=8.58,P<0.01)。>65岁患者与≤65岁患者的突变检出率差异无统计学意义(44.4% vs 44.6%,χ2=0,P>0.05)。肺腺癌的EGFR突变检出率明显高于其他类型的NSCLC,差异有统计学意义(47.7% vs 29.3%,χ2=4.68,P<0.05)。临床I～II期患者的EGFR突变检出率与III～IV期的相比差异无统计学意义(35.0% vs 45.7%,χ2=0.79,P>0.05)。结论 组织样本EGFR基因突变的检出率与胸腔积液样本相比无统计学差异,但明显高于血浆样本,提示临床选择送检的样本类型时,可优先选择组织和胸腔积液样本。EGFR基因突变的检出率与性别和肿瘤的病理类型相关。     

吉非替尼对前列腺癌DU145细胞生长增殖及表皮生长因子受体蛋白表达的影响

目的：观察吉非替尼（Gefitinib）对前列腺癌DU145细胞的生长抑制作用及对细胞表皮生长因子受体（EGFR）蛋白表达水平变化的影响。方法：不同浓度的Gefitinib（020μmol／L）分别作用于体外培养的前列腺癌DU145细胞24～120h后,采用噻唑蓝（MTT）比色法检测细胞生长抑制率,流式细胞仪测细胞周期分布,蛋白免疫印迹（Westernb1ot）检测细胞EGFR蛋白表达水平。结果：Gefitinib可以显著抑制DU145细胞的生长,呈时间一剂量依赖效应,并且细胞生长多停滞于G0／G1期,细胞EGFR蛋白表达分别下降了10．92％（2．5μmol／L）、43．71％（5μmol／L）、53．53％（10μmol／L）和85．98％（20umol／L）,差异有统计学意义（P〈0．05）。结论：Gefitinib能显著抑制前列腺癌DU145细胞的生长,其机制可能与细胞生长在G0／G1期阻滞及EGFR蛋白表达水平下调等因素有关。     

血浆EGFR水平与接受放疗的非小细胞肺癌患者总生存时间相关性研究

【目的】探讨接受胸部放疗的非小细胞肺癌（NSCLC）患者外周血中可溶性表皮生长因子受体（EGFR）水平与患者总生存时间（OS）的相关性。【方法】选择111例局部晚期的 NSCLC 患者,均接受单纯胸部放疗或者同步放化疗,并于放疗开始前采集外周静脉血,采用 ELISA 法检测血浆可溶性 EGFR 水平。治疗完成后定期随访生存状况。【结果】所有患者放疗前血浆可溶性 EGFR 水平为34．9（17．5～70．2） ng／mL ,高于此中位浓度的患者的中位 OS 为30．0个月,低于此中位浓度的患者的中位 OS 为10．1个月,EGFR 水平高者中位 OS 显著高于水平低者,差异具有统计学意义（ P ＜0．01）。多因素分析显示不吸烟／既往吸烟,腺癌,同步放化疗及基线 EGFR ≥34．9 ng／mL 是 OS 延长的独立预后因素（均 P ＜0．05）。【结论】血浆可溶性 EGFR可作为判断接受胸部放疗的局部晚期 NSCLC 患者预后的一项生物学标记物。     

肺腺癌患者表皮生长因子受体酪氨酸激酶结构域突变与放射敏感性的关系

【目的】探讨肺腺癌患者表皮生长因子受体酪氨酸激酶结构域（EGFR-TKD）突变与放射敏感性的关系。【方法】将36例肺腺癌并 EGFR-TKD突变者列为观察组,24例无突变者列为对照组。对两组患者肺部原发病灶均采用三维适形放射治疗,放疗结束后1个月复查胸部 CT评价近期疗效。【结果】全部患者均顺利完成治疗,观察组完全缓解（CR）10例,部分缓解（PR）24例,稳定2例,进展0例,近期总有效率（CR＋PR）94．4％。对照组CR 7例,PR 10例,稳定6例,进展1例,近期总有效率70．8％。两组患者近期有效率比较差异有显著性（P＜0．05）。【结论】肺腺癌 EGFR-TKD突变患者的近期放疗效果好,提示 EGFR-TKD突变是预测放射敏感性的重要指标。     

急性放射性肺损伤肺组织EGFR表达及视黄酸对其影响

[目的]研究通过视黄酸(RA)对大鼠急性放射性肺损伤的干预作用,阐明其对放射性肺损伤肺组织表皮生长因子受体(EGFR)表达的影响.[方法]健康雄性Wistar大鼠80只随机分为4组:正常对照组(A组)、单纯给药组(B组)、单纯照射组(C组)和照射加给药组(D组),每组20只.C、D两组大鼠行6MV-X线15Gy全胸野照射,B、D两组大鼠给予20mg/(kg·d)RA制剂灌服.于照射后第1、2、4、8周取右肺组织行HE和Masson染色,免疫组化方法检测EGFR的表达.[结果]HE和Masson染色提示照射后的1周始肺泡腔有炎性细胞渗出,继之间质水肿.4及8周出现肺泡腔变小甚至结构破坏,局部实变,肺间质出现胶原纤维;B组与A组比各时间段的病理变化无明显差别,但D组与C组比较大鼠肺炎和肺水肿减轻,肺组织胶原纤维量减少.EGFR免疫组化染色显示C组与A组相比在各时间段EGFR表达均明显增强(P<0.001),D组与C组比第4、8周EGFR表达明显减弱(P<0.001).[结论]EGFR参与放射性肺损伤的发生、发展,而RA从蛋白质水平有效抑制EGFR表达,为治疗放射性肺损伤提供了实验依据.     

免疫检查点抑制剂治疗晚期非小细胞肺癌的预后标志物动态监测研究进展

免疫检查点抑制剂在晚期非小细胞肺癌中的应用显著提高了患者的生存获益,但存在治疗反应率低等临床问题。免疫检查点抑制剂治疗是肿瘤-免疫系统交互对话、动态变化的过程,纵向监测生物标志物变化,有助于消除个体间的差异,提供肿瘤时空异质性信息。本文将从肿瘤细胞源性、免疫微环境源性及多指标联合三方面对免疫检查点抑制剂治疗晚期非小细胞肺癌预后标志物动态监测的相关研究进展进行综述。     

血管内皮生长因子联合血清肿瘤标志物对肺癌早期诊断意义

目的探讨血管内皮生长因子（VEGF）联合血清肿瘤标志物对肺癌早期诊断意义。方法 2008年1月-2009年8月收治的92例患者中肺癌患者64例,采集静脉血清标本采用酶联免疫法检测其VEGF水平。结果 64例肺癌与28例非肺癌患者VEGF表达水平分别为（255.72±566.00）、（299.46±795.8）pg/mL,两者比较无统计学意义（P〉0.05）;43例中晚期肺癌VEGF值（125.07±68.2）pg/mL,表达显著高于12例早期肺癌（196.00±260.60）pg/mL（P〈0.05）;CEA与CYFRA21-1的表达对判断26例发生胸膜转移的肺癌有统计学意义（P〈0.05）。结论结合血清VEGF水平和常规肿瘤标志物,可评估现状及临床分期,VEGF结合CEA与CYFRA21-1表达水平为预测肺癌患者早期发生胸膜转移提供理论依据。     

肺癌患者血清VEGF含量测定及临床意义

目的探讨血清血管内皮生长因子在肺癌诊断、预后及疗效观察中的意义.方法用双抗夹心ELISA方法测定50例肺癌患者血清的VEGF水平,另用40名健康人作对照.用SPSS 10.0统计软件包进行统计分析.结果肺癌患者血清VEGF水平显著高于健康人(P<0.001);晚期肺癌(Ⅲ期、Ⅳ期)患者血清VEGF水平高于早期肺癌(Ⅰ期,Ⅱ期)患者,差异有显著性(P<0.01),有淋巴结转移者高于无淋巴结转移者(P<0.05).结论血清VEGF可能成为肺癌的一个新的肿瘤标志物,用于肺癌的诊断和动态监测病情进展、预后判断.     

Mutation Detection of Epidermal Growth Factor Receptor and KRAS Genes Using the Smart Amplification Process Version 2 from Formalin-Fixed,Paraffin-Embedded Lung Cancer Tissue

Recent evidence indicates that the presence of epidermal growth factor receptor (EGFR) or KRAS mutations in non-small cell lung cancer (NSCLC) can predict the response of the tumor to gefinitib. However, it is difficult to detect these mutations using formalin-fixed, paraffin-embedded (FFPE) tissues because the fixation process and aging can damage the DNA. In this study, we describe our work in adapting the Smart Amplification Process version 2 (SmartAmp2) to detect EGFR or KRAS mutations in DNA extracted from FFPE tissues. We were able to detect these mutations in 37 (97%) of 38 FFPE lung cancer tissue samples within 60 minutes with the SmartAmp2 assay and to confirm the correlation between EGFR mutations in FFPE tissues and gefitinib responsiveness. All mutations had previously been confirmed in the 38 samples using DNA extracted from frozen tissues. Electrophoresis results indicated that PCR analysis was not reliable for DNA extracted from FFPE tissue when primers with a long amplicon (>300 bp) were used. This study confirms that the SmartAmp2 assay is suitable for use with DNA extracted from FFPE as well as frozen tissues.Lung cancer, which is responsible for 1.18 million deaths annually worldwide, is the most common cause of cancer mortality in men and the second most common cause in women.¹ Treatment involves a combination of surgery, chemotherapy, and radiation therapy determined based on histological results obtained in biopsy of cancer cells from the individual patient. Despite the greater availability of treatment and substantial research efforts, the prognosis for lung cancer remains poor, and the development of more effective treatments is one of the most important topics in oncology today.Recent studies have indicated that mutations in the epidermal growth factor receptor (EGFR) gene and KRAS gene help physicians decide the course of chemotherapy in patients with non-small cell lung cancer (NSCLC). EGFR mutations^2,3,4,5 and KRAS mutations^6,7,8,9,10 occur in 8 to 10% and 33% of NSCLC patients, respectively, and in 27 to 56% and 5 to 15% of East Asian NSCLC patients, respectively. They are negatively correlated in NSCLC such that patients who have a mutation in the tyrosine kinase domain of the EGFR respond to tyrosine kinase inhibitors such as gefitinib and erlotinib, whereas patients with mutations of the KRAS gene do not respond to this treatment.^3,9,11 Consequently, NSCLC patients with EGFR mutations have a favorable prognosis,^12,13 whereas the prognosis for those with KRAS mutations is poor.^14,15,16,17 Therefore, to provide the optimal therapy, physicians must be able to determine whether patients have EGFR or KRAS gene mutations.Many methods used to detect EGFR or KRAS mutations in clinical samples include restriction fragment length polymorphism,¹⁸ single-strand conformation polymorphism,¹⁹ PCR sequencing,²⁰ high-resolution melting analysis,^21,22 and Scorpions Amplified Refractory Mutation System.²³ All of these methods require careful DNA extraction and purification, involve many steps, and must be performed by skilled technicians. Some of these methods are more sensitive than simple sequencing but are unsuitable for routine clinical use because of their complexity and long turnaround times.Recently, Mitani et al²⁴ developed a rapid, simple, and sensitive mutation detection assay called the Smart Amplification Process version 2 (SmartAmp2). This assay has shown the ability to detect mutations in samples containing as little as 1% mutant allele.^25,26 This assay can be used in the clinical setting, and it allows for the detection of EGFR and KRAS gene mutations within 60 minutes (including sample preparation) and enables high-throughput screening.There are a number of archival formalin-fixed, paraffin-embedded (FFPE) tissue banks worldwide. FFPE tissue is relatively cheap, is easy to ship and handle, provides superior morphological quality, and is compatible with nearly all relevant immunohistochemical antibodies. Consequently, most surgical specimens are stored in FFPE tissue for later analysis of gene mutations if necessary. However, it is time-consuming to extract DNA from FFPE tissue and often difficult to detect mutations because the fixation process and aging can damage DNA.^20,27 The present study demonstrates a technique for adapting the SmartAmp2 method to detect mutations from FFPE tissue. The procedure can detect mutations with high accuracy, and unlike any other method, it gives a reliable diagnostic result based exclusively on amplification. The SmartAmp2 assay provides reliable information from old specimens and marks a major advance in cancer diagnostics.     

人类表皮生长因子受体2在乳腺癌表达与临床的相关病理特征分析

目的 评估人类表皮生长因子受体2(HER2)预测价值,并分析其与常见的组织病理学参数的相关性。方法 收集陕西省人民医院2011年～2014年之间160例接受手术治疗的乳腺癌患者组织标本,通过免疫组织化学方法(IHC)和荧光原位杂交(FISH)检测 HER2水平,通过χ²检验用来评估HER2基因扩增状况及不同临床病理特性的相关性,临床参数包括:肿瘤大小、组织学分级、雌激素受体(ER)和孕激素受体(PR)表达,年龄、绝经情况和Ki-67指数。结果 HER-2表达与组织学分级、淋巴结转移、ER水平、PR,Ki-67指数差异存在统计学意义(均P<0.05)。相对于HER-2⁺组患者,HER-2^-病变多表现为雌激素ER阴性、孕激素阴性、ER阴性、淋巴结阴性、ki-67≥20%。结论 HER-2表达与多种临床病理因素存在相关性。     

乳腺癌组织中血管内皮生长因子与雌激素受体、孕激素受体及人类表皮生长因子的关系

目的探讨乳腺癌组织中人类表皮生长因子(VEGF)与雌激素受体(ER)、孕激素受体(PR)、人类表皮生长因子(Her-2)的关系,进而分析VEGF在乳腺癌各分子分型中的表达是否有差异性。方法应用免疫组化方法检测50例病理证实乳腺癌组织中VEGF、ER、PR、Her-2的表达。结果50例乳腺癌患者中VE