肺腺癌中EGFR突变特异性抗体的表达及其意义

目的：探讨表皮生长因子受体（ epithe1ia1 growth factor receptor,EGFR）突变特异性抗体在肺腺癌中的表达及其意义。方法应用免疫组织化学（ immunohistochemistry,IHC）法检测171例已知EGFR突变情况[扩增阻滞突变系统-PCR（ amp1ification refractory mutation system-PCR,ARMS-PCR）法检测]的肺腺癌中EGFR基因19、21号外显子突变特异性蛋白（ EGFR-19、EGFR-21）和非突变蛋白（ EGFR-P）的表达,分析EGFR-19、21表达与临床病理特征的关系,检验IHC与ARMS-PCR法检测结果的一致性。结果 EGFR突变蛋白表达与低分化腺癌（微乳头型、实体型）有关（ P=0.021）;EGFR-21高表达与肿瘤侵犯胸膜有关（P=0.005）。IHC法检测EGFR-19、21表达与ARMS-PCR法检测EGFR突变具有一致性（Kappa值均>0.4）;EGFR-19、21的敏感性、特异性分别为65.0%、89.4%和70.0%、97.6%。结论肺腺癌中EGFR突变蛋白表达与组织分化差、胸膜侵犯有关,提示其可作为预后不良的指标;IHC法检测EGFR突变特异性蛋白可用作EGFR突变初筛手段。     

肺腺癌EGFR基因19、21号外显子突变分析

目的 探讨肺腺癌表皮生长因子受体(EGFR)基因19、21号外显子突变特点及其临床意义.方法 对肺腺癌手术组织提取DNA,采用聚合酶链式反应(PCR)和直接测序法检测EGFR基因19号与21号外显子突变状态,分析与临床病理参数之间的相关性.结果 在54例被测肺腺癌肿瘤中,发现29例患者存在EGFR基因突变,占53.7％ (29/54).其中,19号外显子突变13例(24.1％),均为缺失突变;21号外显子突变病例为16例(29.6％),均为L858R点突变.高、中分化肿瘤的EGFR突变率均达60％以上,明显高于低分化肿瘤突变率44％.结论 肺腺癌存在较高频率的EGFR基因突变,在高、中分化肿瘤的突变率有高于低分化肿瘤的趋势.     

心包积液中发现肺腺癌EGFR双突变1例

患者女性,80 岁,2017 年4月无明显诱因下出现胸闷、气短、乏力,并伴咳嗽、咳痰,痰液呈白色黏液,20 天后患者自述胸闷症状逐渐加重,至当地医院查胸部CT示心包积液,为求进一步治疗来我院就诊.自诉7 个月前在当地医院查胸部CT示左下肺占位,两肺转移瘤可能,接受当地医院治疗(具体不详).入院后行CT胸部平扫发...     

肺腺癌胸水细胞块EGFR基因突变检测分析

目的探讨肺腺癌胸水细胞块中表皮生长因子受体(epidermal growth factor receptor,EGFR)基因突变检测的临床价值。方法采用ARMS-PCR法检测143例肺腺癌细胞块和269例肺腺癌组织块中EGFR的29种突变类型,并检测细胞块同时送检组织块的患者49例的一致性。结果细胞块EGFR突变型71例,阳性率49.65%(71/143);组织块EGFR突变型131例,阳性率44.26%(131/269);49例有组织块对照的细胞块EGFR结果一致性有36例,一致率达73.47%(36/49),其中细胞块EGFR的阳性率44.90%(22/49),组织块阳性率67.34%(33/49)。结论肺腺癌胸水细胞块EGFR的阳性率略高于组织块;有恶性胸水的肺腺癌患者原发灶组织发生EGFR突变的概率较高。     

肺腺癌患者恶性胸腔积液中EGFR突变状况分析

目的 探讨原发性肺腺癌患者恶性胸腔积液中表皮生长因子受体(epidermal growth factor receptor,EGFR)突变情况.方法 收集恶性胸腔积液标本109例,同时选取该患者的纤维支气管镜或经肺穿刺活检晚期标本109例,提取DNA后,采用RT-PCR法和基因测序法同时进行EGFR 19、21外显子突变检测.结果 两种标本中EGFR突变率分别为56.88%、53.21%,胸腔积液中的突变率与晚期患者活检组织标本相比无明显差异(P=0.35).结论 胸腔积液中的EGFR突变率与其活检组织突变率接近,在晚期肺癌患者临床活检取材困难时,胸腔积液可以作为检测EGFR突变的另一种资源.     

EGFR突变肺腺癌同侧乳腺转移1例

<正>患者女性,52岁。无明显诱因出现咳嗽咳痰、痰中带血,伴胸痛1个月。胸部CT示右肺上叶见团块状软组织影,大小3.6 cm×3.0 cm, 边缘见毛刺及胸膜牵拉;两侧肋骨、肩胛骨、胸骨体及部分腰椎骨质破坏;右侧乳腺有一肿块,大小2.4 cm×1.3 cm, 右侧腋窝及纵隔内见肿大淋巴结。行右侧乳腺肿块穿刺活检,病理报告：(右侧乳腺)浸润性导管癌,Ⅲ级伴大汗腺化生。     

应用ARMS检测不同类型的肺腺癌标本EGFR基因突变

目的：探讨应用扩增阻滞突变系统（ amp1ification re-fractory mutation system,ARMS）法检测不同类型肺腺癌标本表皮生长因子受体（ epithe1ia1 growth factor receptor,EGFR）基因突变的特点。方法收集80例山东省西部地区肺腺癌患者肿瘤标本（胸水细胞块、肺活检、手术切除组织）,采用ARMS法检测EGFR基因19、20、21号外显子突变状态,并分析EGFR突变与标本类型、患者性别、年龄、吸烟状态及家族史等的关系。结果肺腺癌中,总EGFR突变率为41.25%,胸水细胞块、肺活检、手术切除组织 EGFR 突变率分别为43.14%、31.25%、46.15%（ P =0.649）。EGFR 突变标本中,21号外显子错义突变（L858R）发生比例最高（59.58%,19/33）,其次为19号外显子缺失（19de1）（39.39%,13/33）,二者同时突变占3.03%（1/33）;女性与男性的突变率差异有显著性（54.55% vs 25.00%,P=0.008）;吸烟与不吸烟患者的突变率差异有显著性（25.81% vs 51.02%,P=0.026）。结论在肺腺癌中,胸水细胞块、肺活检、手术切除组织等标本均可用于EGFR基因突变检测,21号外显子L858R突变最常见;女性、不吸烟患者突变率明显高于男性、吸烟患者。     

四川地区肺腺癌中EGFR基因19、21外显子突变研究

目的探讨EGFR基因19、21外显子突变与肺癌组织学类型、人种、年龄、性别、是否吸烟和所在地域的关系。方法收集65例四川地区肺腺癌患者石蜡包埋组织样本,选用针对19、21外显子的野生型和突变型基因的特异性引物,采用等位基因特异性寡核苷酸PCR法(allele-specific oligonucleotide polymerase chain reaction,ASO-PCR)检测突变。结果 65例肺腺癌样本中,35例发生19、21外显子突变(53.85%),其中19外显子缺失突变13例(20%),21外显子L858R错义突变23例(35.38%);男性38.24%(13/34),女性70.97%(22/31);吸烟者31.25%(5/16),非吸烟者61.22%(30/49);≤60岁的患者为57.14%(16/28),>60岁的患者为51.35%(19/37)。结论 65例四川地区原发肺腺癌中EGFR基因19、21外显子突变在女性、非吸烟患者中更常见(P<0.05),其中21外显子L858R突变率高于19外显子缺失率(P<0.05)。与文献报道比较,肺腺癌EGFR基因总突变率较其他类型肺癌高,且高于白种人。     

非小细胞肺癌中EGFR突变位点和样本差异性对突变与蛋白表达的影响

目的探讨非小细胞肺癌(non-small cell lung cancer,NSCLC)中表皮生长因子受体(epidermal growth factor receptor,EGFR)基因突变的临床病理特征,分析EGFR突变与蛋白表达的相关性以及不同突变位点和样本差异性的影响,寻求更有效的EGFR筛查手段。方法采用ARMS-PCR法和免疫组化法检测356例NSCLC中EGFR基因突变和蛋白的表达,分析两者的相关性。结果EGFR基因突变率为53. 93%,EGFR蛋白阳性率为69. 8%;两者表达显著相关(P0. 001);以ARMS-PCR法为金标准时,免疫组化法检测其灵敏度为87. 77%,特异性为53. 77%,阳性预测值为71. 34%,阴性预测值为77. 02%;因标本类型和组织学类型造成的样本差异性中,手术肿块(Kappa=0. 824)和肺鳞癌(Kappa=0. 496)样本的检测一致性明显优于其他组织; EGFR突变与患者性别(P 0. 001)、组织学类型(P 0. 001)、胸膜侵犯(P 0. 001)、脉管神经累犯(P=0. 001)和淋巴结转移(P=0. 002)密切相关。结论非小细胞肺癌中EGFR突变与蛋白表达相关,在充分考虑样本差异性影响的前提下,免疫组化检测可作为EGFR突变的经济便捷初筛手段和补充。     

Monitoring of cyclooxygenase-2 levels can predict EGFR mutations and the efficacy of EGFR-TKI in patients with lung adenocarcinoma

Background: Epidermal growth factor receptor (EGFR) mutation detection has become a routine molecular test with significant implications for prognosis and therapeutic options of EGFR tyrosine kinase inhibitors (EGFR-TKIs). However, acquiring sufficient amounts of tissue for analyzing EGFR mutations is not often feasible, and not all the patients with sensitive EGFR mutations have benefit from EGFR-TKI treatment. Method: EGFR mutations were detected by amplification refractory mutation system (ARMS) in 44 patients of newly diagnosed lung adenocarcinoma, and patients with EGFR-positive mutations received EGFR-TKI treatment. The serum cyclooxygenase-2 (COX-2) levels were tested before EGFR-TKI treatment and on the 30th days after EGFR-TKI treatment. Results: Twenty-nine cases were detected EGFR mutations. EGFR mutation rate of serum COX-2 high-level group was significantly higher than low-level group (92.9% vs. 53.3%, P = 0.025). Multivariate analysis showed that serum COX-2 level was independently associated with EGFR mutation (P = 0.033, OR = 12.385, 95%CI, 1.231-124.567). Analysis of the correlation between clinical characteristics and the response of EGFR-TKI showed that the serum COX-2 high-level group had a better efficacy than low-level group (P = 0.000), and multivariate logistic regression analysis showed that the serum COX-2 level was the independently influencing factor (P = 0.004). Kaplan-Meier analysis showed that patients of COX-2 high-level group have longer progression-free survival (PFS, P = 0.013), and the Cox regression analysis showed that the same result (P = 0.003; OR = 0.980, 95% CI, 0.967-0.993). Conclusion: The serum COX-2 level seems to be closely associated with EGFR mutations in patients with Lung adenocarcinoma. The serum COX-2 level could help us to predict the responses of EGFR-TKI and the PFS in patients harboring EGFR mutation.     

免疫组化法检测恶性胸腔积液脱落细胞EGFR突变状态

目的：评估免疫组织化学染色法(immunohistochemistry,IHC)在恶性胸腔积液脱落细胞EGFR突变检测中的应用价值。方法：连续纳入我院2012年1月至2013年12月154例肺腺癌患者,筛选合格病例入组,使用扩增阻滞突变系统技术(amplification refractory mutation system,ARMS)和IHC分别检测肺癌组织和胸腔积液脱落细胞的EGFR突变状态。比较两种方法的检测敏感度、特异度和一致率等。结果：85例患者最终纳入本研究,患者平均年龄66.4岁。ARMS法检测肿瘤组织的总体突变率约49.4%,其中E746_A750 del突变率17.6%,L858R突变率22.4%。以ARMS法为金标准,IHC染色检测E746_A750 del和L858R突变的敏感度分别为93.3%和89.5%,两种方法的检测一致性极好。进一步,85例患者胸腔积液的脱落肿瘤细胞检出率为41.2%,IHC法检测E746_A750 del和L858R突变的敏感度分别为83.3%和66.7%,特异度均为100.0%。结论：IHC染色是一种可靠、准确的EGFR突变检测方法,能够获得良好的特异度和一致率,可用于恶性胸腔积液脱落肿瘤细胞的常规筛查。     

EGFR and KRAS mutations in metastatic lung adenocarcinomas

In primary lung adenocarcinoma, EGFR and KRAS mutations are found in approximately 10% to 20% and 20% to 30%, respectively. Few studies have investigated these mutations in metastases. Patients with EGFR mutations have a 70% to 80% response rate to tyrosine-kinase inhibitors therapy and a longer progression-free survival rate in contrast to patients with KRAS mutations that are associated with virtually no response tyrosine-kinase inhibitors. In this study, we have investigated EGFR and KRAS mutations in metastatic lung adenocarcinoma. Using Johns Hopkins Hospital archives, 1966 lung adenocarcinomas were found from January 2007 to May 2010. A total of 60 metastatic adenocarcinomas (28 cytologic and 32 surgical cases) with EGFR and KRAS studies were identified. In addition, 18 cases of primary and matched metastases were also included. Exons 18 to 21 of EGFR and exon 2 of KRAS (codons 12 and 13) were sequenced. In our study, EGFR and KRAS mutations were found in 21.7% (13 of 60 cases) and 28.3% (17 of 60 cases), respectively, and occurred more often with advanced stage of primary tumors. KRAS mutations were associated with poor prognosis and occurred exclusively in smokers in comparison with EGFR mutation. Of 9 pairs, mutations were concordant in 77.8%; 1 pair displayed acquisition of KRAS mutation, whereas 1 pair showed loss of EGFR mutation in the corresponding metastasis. Our findings suggest that EGFR and KRAS status should be tested in metastasis regardless of known mutations of the primary tumor. Additional studies are needed to further investigate the mechanisms of discordances in metastatic tumors.     

A comparison of ARMS and mutation specific IHC for common activating EGFR mutations analysis in small biopsy and cytology specimens of advanced non small cell lung cancer

We have compared mutation analysis by Amplification Refractory Mutation System (ARMS) and epidermal growth factor receptor (EGFR) mutant-specific antibodies for their ability to detect two common activating EGFR mutations in a cohort of 115 advanced non-small cell lung cancer (NSCLC), including cytology material, core biopsy, and bronchoscopic biopsies. Assessment of EGFR mutation status was performed by using antibodies and ARMS assay specific to the two major forms of mutant EGFR, exon 19 deletion E746-A750 (c.2235_2249del15 or c.2236_2250del15, p. Glu746_Ala750 del) and exon 21 L858R point mutation (c.2573T>G, p.Leu858Arg). In this study the optimal buffer for antigen retrieval was sodium citrate (pH 6.0). Q score was used to evaluate the specific mutant EGFR proteins expression. Validation using clinical material showed deletions in exon 19 were detected in 19.1% and L858R mutation in 20% of all cases by ARMS assay. A cutoff value of score 1 was used as positive by IHC. No wild type cases were immuno-reactive. The antibodies performed well in cytology, core biopsies and bronchoscopic biopsies. There were only one false positive case using L858R IHC (sensitivity 100%, specificity 98.5%, positive predictive value 96%, negative predictive value 100%). All 23 E746-A750 exon 19 deletions identified by mutation analysis were positive by IHC. The sensitivity of exon 19 IHC for E746-A750 was 100%, specificity 100%, positive predictive value 100% and negative predictive value 100%. The result of the IHC stains was finely correlated with mutations status determined by ARMS assay. Although inferior to molecular genetic analysis of the EGFR gene, IHC is highly specific and sensitive for the targeted EGFR mutations. The antibodies are likely to be of clinical value in cases especially where limited tumor material is available, or in situations where molecular genetic analysis is not readily available.     

Correlation between EGFR gene mutation pattern and Akt phosphorylation in pulmonary adenocarcinomas

The detection of gene mutation of epithelial growth factor receptor (EGFR) is important to predict the therapeutic effect of gefitinib. Recently, it was reported that examination of the activation of the downstream protein of EGFR is useful in the same way as the EGFR mutation. Therefore the purpose of the present paper was to determine whether activation of Akt and Erk, which are downstream proteins, and the EGFR gene mutation pattern was correlated. A total of 130 pulmonary adenocarcinomas were studied for the gene mutations of EGFR in exon 19 and 21, and the phosphorylation of Akt and Erk was investigated by immunostaining. The EGFR mutation was detected in 32%, the positivity of p-Akt was 51%, and the rate of p-Erk was 27%. The EGFR mutation-positive cases were the minority in p-Akt-negative cases, and the p-Akt expression was significantly associated with the mutation of EGFR (P=0.0014). In addition, there was a significant correlation between the L858R mutation and the expression of p-Akt (P=0.040). It is suggested that the activation of Akt is dependent on EGFR mutation pattern.