EGFR 突变种类与临床疗效关联

癌组织中表皮生长因子受体(epidermal growth factor receptor,EGFR)基因突变是应用靶向药物EGFR酪氨酸激酶抑制剂(tyrosine kinase inhibitor,TKI)治疗的一个重要相关因素及预测指标。对其突变的检测可以指导TKI类药物(TKIs)的最佳应用。该种突变常出现在非小细胞肺癌(NSCLC)中,尤其是在亚洲女性、肺腺癌、非吸烟者中,与非小细胞肺癌患者对TKIs治疗的敏感性密切相关。本文旨在探讨利用EGFR基因的已知突变热点的相关知识选择适合不同分子遗传学背景的群体或/和个体的"个体化"治疗方案,最终达到延长肺癌患者生存时间和提高生活质量的双重目的。     

肺癌EGFR突变与酪氨酸激酶抑制剂临床敏感性的关系

表皮生长因子受体(EGFR)酪氨酸激酶抑制剂(TKI)是近年来在临床中使用的一类新的小分子靶向药物,主要用于晚期非小细胞肺癌(NSCLC)的治疗,然而并非所有的NSCLC患者对TKI敏感。近期研究发现,在NSCLC治疗过程中,EGFR突变与TKI临床敏感性密切相关,通过检测肺癌EGFR突变状况可以预测TKI治疗的效果。     

肺癌的表皮生长因子受体分子靶向治疗与基因突变

肺癌分子靶向治疗近年来取得较大进展,特别是针对表皮生长因子受体（EGFR）分子靶向药物表现出确定的临床效果。临床应用表明,EGFR基因酪氨酸激酶域体细胞突变与非小细胞肺癌患者对酪氨酸激酶抑制剂吉非替尼的敏感性相关,本文就相关的研究进行了简述。     

基于液体活检与二代测序解析肺腺癌患者靶向药耐药机制

表皮生长因子受体酪氨酸激酶抑制剂(EGFR TKI)是治疗非小细胞肺癌的一线药物,但在治疗过程中难以避免耐药性的产生.目前已知的获得性耐药机制主要包括EGFR T790M突变、HER2基因扩增、MET基因扩增、转分化等,并尚有15％～20％的患者的耐药机制不明.本研究发展了一种基于液体活检与高通量测序方式解析肺癌患者靶向药物耐药机制的方法.我们通过液体活检的方式取得了接受EGFR TKI治疗肺腺癌患者治疗前及EGFR TKI耐药后的胸腔积液样本,在对其中的肿瘤细胞进行富集后,通过基因组及转录组的高通量测序并结合生物信息学分析,解析耐药前、耐药后基因组变异以及基因表达的差异,进而探究该患者的靶向药物耐药机制,为制订新的治疗方案提供科学依据.     

EGFR突变与替莫唑胺联合IGRT大分割放射治疗非小细胞肺癌脑转移瘤疗效的相关性

摘要 目的：研究表皮生长因子受体（EGFR）突变对替莫唑胺联合图像引导大分割放射（IGRT）治疗非小细胞肺癌脑转移瘤临床疗效的影响。方法：选择2015年1月到2018年12月在我院接受治疗的非小细胞肺癌脑转移患者86例,根据是否出现EGFR突变分为对照组（EGFR未突变组）和研究组（EGFR突变组）,每组43人,两组患者均接受替莫唑胺联合IGRT大分割放射治疗。比较两组患者临床治疗疗效、不良反应发生情况、复发时间、生存时间和生活质量。结果：研究组患者临床治疗总有效率较对照组患者高（P<0.05）。研究组患者治疗后复发时间和生存时间均显著高于对照组患者（P<0.05）。两组患者治疗期间头痛、恶心、疲乏以及神经毒性等不良反应的发生情况比较无显著差异（P>0.05）。两组患者治疗前生活质量KarnofSky活动状态评分（KPS）和肺癌相关症状量表（LCSS评分）无显著差异（P>0.05）;治疗后,研究组患者KPS评分显著高于对照组（P<0.05）,而LCSS评分显著低于对照组患者（P<0.05）。结论：替莫唑胺联合IGRT大分割放射治疗EGFR突变的非小细胞肺癌脑转移瘤临床疗效更好,并且治疗后患者生活质量更优。     

非小细胞肺癌EGFR-TKIs治疗及PET分子成像的研究进展

表皮生长因子受体(epithelial growth factor receptor,EGFR)信号转导通路在非小细胞肺癌(Non-Small Cell Lung Cancer,NSCLC)中发挥重要作用,尤其胞内酪氨酸激酶结构域的突变状态决定了目前NSCLC的靶向治疗。针对EGFR突变的分子靶向药物表皮生长因子受体酪氨酸激酶抑制剂(epithelial growth factor receptor tyrosine kinase inhibitors,EGFR-TKIs)已开发并应用于NSCLC的治疗。在治疗过程中,EGFR的突变状态随时间发生动态变化,因此精准掌握EGFR的突变状态是靶向治疗方案制定、优化的关键。PET分子成像可在细胞和分子水平,对在体生物活动的发生、发展过程进行实时成像,使实时、在体揭示EGFR的突变状态成为可能。因此,多种以TKIs为前体标记放射性核素作为靶向肿瘤突变EGFR胞内段分子成像探针的研究逐渐增多。本文就EGFR-TKIs在NSCLC治疗及相关PET分子成像方面的研究进展进行综述。     

表皮生长因子受体基因突变对非小细胞肺癌脑转移患者生存期的影响

为了探究表皮生长因子受体(epidermal growth factor receptor,EGFR)基因的突变对非小细胞肺癌(non-small cell lung cancer,NSCLC)脑转移患者生存期的影响,我们以2012年1月至2014年12月期间我院收治的123例非小细胞肺癌脑转移患者为回顾性研究对象,按治疗方案的差异分为放射治疗组(36例),化疗组(52例),放射治疗+化疗组(24例)、放化疗+靶向治疗组(13例),观察EGFR基因突变对不同治疗方案的非小细胞肺癌脑转移患者无进展生存期(progression-free-survival,PSF)和总生存期(overall survival,OS)的影响。我们发现,EGFR突变对非小细胞肺癌脑转移患者生存期具有显著影响,EGFR突变型患者生存期明显长于EGFR野生型患者,而基于EGFR突变的靶向治疗患者生存期明显长于辅助治疗患者,因此基于EGFR突变的靶向治疗可有效提升患者生存期。我们的研究为非小细胞肺癌脑转移患者的临床治疗提供了一定的理论依据。     

自噬与非小细胞肺癌EGFR-TKI耐药的研究进展

肺癌是目前世界上发病率和死亡率均居首位的恶性肿瘤,其中大多数是非小细胞肺癌(non-small cell lung cancer,NSCLC)。在对NSCLC的治疗中,酪氨酸激酶抑制剂(tyrosine kinase inhibitor,TKI)的应用已成为靶向表皮生长因子受体(epidermal growth factor receptor,EGFR)的重要治疗手段。但由于获得性耐药问题的出现,使得这类靶向药物的作用受到限制。近年来大量研究表明,除了基因突变等分子水平的因素外,细胞自噬(autophagy)在非小细胞肺癌EGFR-TKI靶向药物耐药中也发挥了重要的作用。该文重点就自噬与非小细胞肺癌EGFR-TKI靶向药物耐药的研究进展进行分析,旨在为解决自噬相关的EGFR-TKI耐药提供更多思路。     

基因突变检测技术在非小细胞肺癌表皮生长因子受体 与临床病理特征的关系研究中的作用

摘要：目的 探讨非小细胞肺癌（NSCLC）表皮生长因子受体（EGFR）基因突变情况及其与临床病理的关系。方法 选取我院2010年1月至2014年1月收集的159份非小细胞肺癌手术切除标本为研究对象,利用基因测序方法检测标本中的EGFR基因突变情况,并分析其与临床病理的关系。结果 159例样本中,EGFR基因突变检出率为12.6%(20/159),突变主要集中在19号外显子的缺失和21号外显子的点突变。女性患者基因突变检出率明显高于男性患者（P﹤0.01）。腺癌及细支气管肺泡癌患者基因突变检出率明显高于其他组织学分型（P﹤0.01）。高分化患者基因突变检出率高于中－低分化检出率（P＜0.05）。EGFR基因突变与年龄及淋巴结转移与否无关（P＞0.05）。结论 非小细胞肺癌患者EGFR基因突变与性别、组织学分型及分化程度密切相关。     

EGFR基因在非小细胞肺癌、乳腺癌中突变的研究

表皮生长因子受体(EGFR)基因酪氨酸激酶域体细胞突变与非小细胞肺癌(NSCLC)患者对酪氨酸激酶抑制剂吉非替尼敏感性密切相关。文章分析和检测本院75例非小细胞肺癌、10例乳腺癌患者石蜡包埋标本EGFR基因突变状况。采用PCR技术进行EGFR基因19和21外显子突变分析。结果显示:75例NSCLC患者中有13例(13/75,17.33%)酪氨酸激酶域存在体细胞突变。其中7例(7/75,9.33%)为19外显子缺失突变,6例(6/75,8%)为21外显子替代突变(2573T>G,L858R)。病理分型显示,腺癌突变率高于其他几种类型NSCLC。乳腺癌患者均为免疫组化HER-2阳性女性,EGFR基因的19、21外显子中未见突变发生。中国非小细胞肺癌患者总突变率高于高加索人种,女性患者较男性患者突变率高,提示肺腺癌的患者突变率高可能在吉非替尼的治疗中获益。     

陕西省南部地区非小细胞肺癌EGFR基因突变分析

目的:探讨陕西南部非小细胞肺癌表皮生长因子受体基因的突变状况。方法:采用测序方法检测陕西省南部地区233例非小细胞肺癌(non-small cell lung cancer,NSCLCs)患者表皮生长因子受体(epithelial growth factor receptor,EGFR)基因第18、19、20和21号外显子突变情况,并分析其基因突变与肺癌人口学分布及组织类型的关系。结果:233例非小细胞肺癌患者中,共检出82例含有EGFR基因突变,其中第18、19、21号外显子突变率分别为1.3%、16.3%和18.0%,第20号外显子无突变;男性EGFR基因突变率(31.2%,39/125)低于女性(39.8%,43/108);腺癌EGFR基因突变率(39.1%,75/192)高于鳞癌(22%,9/41)。结论:陕西南部NSCLC的EGFR基因突变率较高,以第19、21号外显子突变为主。EGFR基因变率与NSCLC患者性别和病理类型均无关。     

胸腔积液中肺腺癌细胞EGFR突变状态与DNA含量的相关性

摘要 目的：探究胸腔积液中肺腺癌细胞表皮生长因子受体（epidermalgrowthfactorreceptor,EGFR）突变状态与DNA含量的相关性,以期探究EGFR突变状态是否同肿瘤的恶性程度存在一定关联。方法：选择2015年1月至2020年1月于我院接受EGFR基因检测以及基因定量分析的591例肺腺癌患者为研究对象,按照其是否出现EGFR基因突变将其分为突变组（335例）与非突变组（256例）,两组患者的胸腔积液均使用激光图像细胞仪开展DNA含量以及非整倍体峰检测,并开展组间差异性比较。结果：（1）将591例患者按照年龄、性别及是否吸烟等临床特征进行分组对比显示,性别（P=0.034）与吸烟（P=0.007）同肺腺癌患者胸腔积液细胞出现EGFR突变具有一定关联,而年龄因素与是否出现突变无明显相关性（P>0.05）;（2）突变组患者的最大DNA指数（DI）、大于5C细胞的平均DI以及大于9C细胞的平均DI均明显高于非突变组,组间差异明显（P<0.05）;（3）开展DNA非整倍体细胞峰比较显示突变组在单峰、双峰占比中明显高于非突变组,而无峰占比明显低于非突变组（P<0.05）,多峰占比方面两组差异不大（P>0.05）。结论：经研究显示,同未出现EGFR突变的肺腺癌患者相比较,发生EGFR突变的肺腺癌患者明显DI值更高,非整倍体细胞以及非整倍体峰值也呈现异常升高态,这提示EGFR发生突变的肺腺癌患者恶变洗吧的侵袭性更强。     

Suitability of Surgical Tumor Tissues,Biopsy, or Cytology Samples for Epidermal Growth Factor Receptor Mutation Testing in Non–Small Cell Lung Carcinoma Based on Chinese Population

BACKGROUND: Epidermal growth factor receptor (EGFR) mutation status is crucial in treatment selection for non–small cell lung cancer (NSCLC) patients; however, the detection materials’ availability remains challenging in clinical practice. In this study, we collected surgical resection tissues, lymph node biopsy, and cytological samples for EGFR mutation testing and investigated the associations between gene mutation and clinical characteristics. METHODS: Two hundred and seventy-six NSCLC adenocarcinoma specimens were collected, and highly sensitive amplification refractory mutation system method was implemented for EGFR mutation detection, with clinicopathologic characteristics involved in the final analysis. RESULTS: In the total of 276 samples, 96% (265/276) of tumors obtained evaluable EGFR mutation status, the frequency of mutation was 55.8% (148/265) in all specimens, and three different type samples shared a comparable successful testing rate: 97.4% (38/39) in surgical tumor tissues, 100% (108/108) in lymph node biopsy samples, and 92.2% (119/129) in cytological samples. EGFR mutation was significantly associated with sex, smoking history, lymph node metastasis status (N stage), primary tumor size, testing tissues origin, and sample type (P < .05). Multivariate analysis reconfirmed that smoking history and primary tumor size shared significant correlation with EGFR mutation after adjustment. CONCLUSIONS: Both lymph node biopsy and cytological samples were suitable surrogates for EGFR mutation detection in NSCLC compared with tumor tissues, gene status should be detected widely considering the high EGFR mutation rate, and nonsmoking history together with smaller primary tumor size was an independent indicator of EGFR mutation status.     

Maintenance of EGFR and EGFRvIII expressions in an in vivo and in vitro model of human glioblastoma multiforme

Glioblastoma multiforme (GBM) is the most common, and most aggressive primary brain tumor among adults. A vast majority of the tumors express high levels of the epidermal growth factor receptor (EGFR) as a consequence of gene amplification. Furthermore, gene amplification is often associated with mutation of EGFR, and the constitutive activated deletion variant EGFRvIII is the most common EGFR mutation found in GBM. Activated EGFR signaling, through overexpression and/or mutation, is involved in increased tumorigenic potential. As such, EGFR is an attractive target for GBM therapy. However, clinical studies with EGFR inhibitors have shown inconsistent results, and as such, further knowledge regarding the role of EGFR and EGFRvIII in GBM is needed. For this, an appropriate in vivo/in vitro tumor model is required. Here, we report the establishment of an experimental GBM model in which the expressions of EGFR and EGFRvIII are maintained both in xenograft tumors growing subcutaneously on mice and in cell cultures established in stem cell conditions. With this model it will be possible to further study the role of EGFR and EGFRvIII, and response to targeted therapy, in GBM.     

基于18F-FDG PET/CT参数评价不同EGFR基因状态晚期肺腺癌患者放化疗疗效预测价值

为评价18F-FDG PET/CT参数对不同表皮生长因子受体(epithelial growth factor receptor, EGFR)基因状态晚期肺腺癌患者同步放化疗的疗效预测价值,本研究选择2016年1月至2018年2月我院初诊的100例晚期肺腺癌患者,比较不同EGFR基因状态下晚期肺腺癌患者同步放化疗治疗的临床疗效,并对18F-FDG PET/CT显像中原发病灶的各项代谢参数,代谢肿瘤体积(MTV)、最大标准摄取值(SUVmax)、平均标准摄取值(SUVmean)和总肿瘤糖酵解(TLG)值,以预测不同EGFR基因状态晚期肺腺癌患者同步放化疗效果的受试者工作特征曲线(receiveroperatingcharacteristiccurve,ROC曲线),并进行分析。结果显示,SUVmax预测不考虑EGFR基因状态的晚期肺腺癌患者放化疗疗效的截断值为9.50 (AUC=0.715,敏感度=79.7%,特异性=61%,p=0.031),SUVmax预测EGFR野生型的晚期肺腺癌患者放化疗疗效的截断值为9.50 (敏感度=82.4%,特异性=64.3%, p=0.014)。MTV预测EGFR 19号及21号外显子突变的晚期肺腺癌患者放化疗疗效的截断值分别为93.50(敏感度=63.6%,特异性=92.3%, p=0.021),77.00 (敏感度=83.3%,特异性=69.2%, p=0.041)。综上所述,对于EGFR基因状态不明的肺腺癌患者,SUVmax值可较好的预测放化疗效果,尤其是对于EGFR野生型患者;当EGFR19号及21号外显子突变时,MTV值预测结果优于SUVmax值。     

Simultaneous detection of multiple mutations in epidermal growth factor receptor based on fluorescence quenching of quantum dots

We have developed a simultaneous detection method for two common mutations in the epidermal growth factor receptor gene based on the fluorescence quenching phenomenon caused by aggregation of CdSe quantum dots. For detection of the in-frame deletion in exon 19 and the L858R point mutation in exon 21, water-soluble CdSe quantum dots with two sizes were functionalized using four different types of probe oligonucleotides. Addition of target oligonucleotides with the deletion mutation in exon 19 into the suspensions caused crosslinking-induced aggregation of green-emitting quantum dots, followed by the fluorescence quenching while that with the L858R point mutation resulted in aggregation of yellow-emitting quantum dots. In addition, targets with both deletion and point mutations caused aggregation of both green- and yellow-emitting quantum dots. This method allows a simultaneous detection of mutations in exon 19 and 21 of EGFR gene in a single experiment. We found that minimum mutant concentration that could be detected by this method was as low as 2% for deletion mutation, and 5% for point mutation. PCR products of EGFR gene were also used to confirm that our method could be used to detect mutation in amplified DNA fragments.     

Methylation of TMEFF2 gene in tissue and serum DNA from patients with non-small cell lung cancer

Lung cancer remains a global health problem with a high mortality rate. CpG island methylation is a common aberration frequently associated with gene silencing in multiple tumor types, emerging as a highly promising biomarker. The transmembrane protein with a single EGF-like and two follistatin domains (TMEFF2) is epigenetically silenced in numerous tumor types, suggesting a potential role as a potential tumor suppressor. However, the role of TMEFF2 in lung cancer remains to be fully elucidated. We explored the methylation status of TMEFF2 gene in 139 patients with non-small cell lung cancer (NSCLC) and the feasibility of detecting circulating methylated DNA as a screening tool for NSCLC using methylation-specific PCR in 316 patients and 50 age-matched health controls. TMEFF2 methylation in tumor tissues was found in 73 of the 139 NSCLCs (52.5%) and was related to gene expression. The frequency of TMEFF2 methylation was higher in females and never-smokers than in males and smokers with borderline significance (65.8% vs 47.8%, p = 0.06; 65.7% vs 48.1%, p = 0.07). Notably, in adenocarcinomas, TMEFF2 methylation was significantly more frequent in tumors without EGFR mutation than those with EGFR mutation (adjusted odds ratio = 7.13, 95% confidence interval = 2.05-24.83, P = 0.002). Furthermore, TMEFF2 methylation was exclusively detected in the serum of NSCLC patients at a frequency of 9.2% (29/316). These findings suggest that methylation-associated down-regulation of TMEFF2 gene may be involved in lung tumorigenesis and TMEFF2 methylation can serve as a specific blood-based biomarker for NSCLC.     

Molecular Epidemiology of EGFR Mutations in Asian Patients with Advanced Non-Small-Cell Lung Cancer of Adenocarcinoma Histology – Mainland China Subset Analysis of the PIONEER study

Epidermal growth factor receptor (EGFR) mutations are the strongest response predictors to EGFR tyrosine kinase inhibitors (TKI) therapy, but knowledge of the EGFR mutation frequency on lung adenocarcinoma is still limited to retrospective studies. The PIONEER study (NCT01185314) is a prospective molecular epidemiology study in Asian patients with newly diagnosed advanced lung adenocarcinoma, aiming to prospectively analyze EGFR mutation status in IIIB/IV treatment-naïve lung adenocarcinomas in Asia. We report the mainland China subset results. Eligible patients (≥20 yrs old, IIIB/IV adenocarcinoma and treatment-naïve) were registered in 17 hospitals in mainland China. EGFR was tested for mutations by amplification refractory mutation system using biopsy samples. Demographic and clinical characteristics were collected for subgroup analyses. A total of 747 patients were registered. Successful EGFR mutation analysis was performed in 741, with an overall mutation rate of 50.2%. The EGFR active mutation rate is 48.0% (with 1.3% of combined active and resistance mutations). Tobacco use (>30 pack-year vs. 0–10 pack-year, OR 0.27, 95%CI: 0.17–0.42) and regional lymph nodes involvement (N3 vs. N0, OR 0.47, 95%CI: 0.29–0.76) were independent predictors of EGFR mutation in multivariate analysis. However, even in regular smokers, the EGFR mutation frequency was 35.3%. The EGFR mutation frequency was similar between diverse biopsy sites and techniques. The overall EGFR mutation frequency of the mainland China subset was 50.2%, independently associated with the intensity of tobacco use and regional lymph nodes involvement. The relatively high frequency of EGFR mutations in the mainland China subset suggest that any effort to obtain tissue sample for EGFR mutation testing should be encouraged.