非小细胞肺癌患者EGFR突变率及与临床病理关系和TKI靶向治疗效果

目的探讨非小细胞肺癌(NSCLC)患者表皮生长因子受体(EGFR)突变率及与临床病理关系和酪氨酸酶抑制剂(TKI)靶向治疗效果。 方法选取2014年5月至2016年8月我院收治的NSCLC患者100例为研究对象,采用聚合酶链反应-直接测序法检测患者肿瘤组织EGFR基因18-21号外显子突变情况,分析EGFR突变与临床病理的关系及其突变特点,比较EGFR突变型与野生型采用TKI靶向治疗的疗效及1年生存率、2年生存率。 结果本次纳入的100例NSCLC患者中共44例发生EGFR突变,EGFR突变率为44.00%;NSCLC患者EGFR突变率特点：女性高于男性,吸烟患者高于不吸烟患者,腺癌高于非腺癌,差异对比均有统计学意义(P<0.05);Logistic回归分析显示性别为女性、病理类型为腺癌是导致NSCLC患者发生EGFR突变的独立危险因素(P<0.05);EGFR突变类型包括18号外显子点突变4.54%(2/44)、19号外显子缺失突变43.19%(19/44)、20号外显子插入突变及点突变11.36%(5/44)、21号外显子点突变40.91%(18/44);EGFR突变型患者TKI治疗有效率68.18%明显高于EGFR野生型10.71%(P<0.05);EGFR突变型与野生型患者1年、2年生存率对比差异无统计学意义(P>0.05)。 结论性别、病理类型是导致NSCLC患者发生EGFR基因突变的独立危险因素,对于EGFR突变型患者采用TKI靶向治疗可获得较好疗效,但2年生存率与EGFR野生型患者尚无明显差异。     

安徽省非小细胞肺癌表皮生长因子受体基因的突变

目的 探讨安徽省地区非小细胞肺癌(NSCLC)患者中表皮生长因子受体(EGFR)基因突变的情况,比较特异引物双扩增系统(ADxARMS)和直接测序检测EGFR基因突变的敏感性.方法 对甲醛溶液固定石蜡包埋的手术标本用显微切割技术获取肿瘤组织细胞,采用QIAamp DNA提取试剂盒提取基因组DNA,通过ADx-ARMS及DNA直接测序技术分析NSCLC患者中EGFR基因突变情况.结果 45例NSCLC中,ADx-ARMS检测有29例存在EGFR基因突变,突变检出率为64.4％,其中22例为外显子19的缺失突变,6例为外显子21点突变,1例为外显子19和21的共同突变.直接测序检测出12例EGFR基因突变与ADx-ARMS检测结果一致.非鳞癌EGFR基因突变率84.8％( 28/33)高于鳞癌患者8.3％(1/12),两者之间的差异具有统计学意(P=0.000);女性患者的突变率为83.3％( 20/24)高于男性患者42.9％(9/21),两者之间的差异具统计学意义(P=0.005).结论 在安徽地区NSCLC中,EGFR基因的突变以外显子19的缺失突变和外显子21的点突变为主,突变率以非鳞癌和女性较高.ADx-ARMS法能快速、准确检测EGFR基因突变.     

重庆地区非小细胞肺癌表皮生长因子受体突变检测与临床病理特征分析

目的探讨重庆市地区非小细胞肺癌（NSCLC）患者表皮生长因子受体（EGFR）突变状态及其与临床病理特征的关系。方法收集55例NSCLC患者的癌组织或胸水标本,采用扩增阻滞突变系统（ARMS）检测EGFR外显子18、19、20和21突变状态。结果55例患者中,EGFR突变14例,突变率25．45％;其中,男、女突变率分别为12．50％、43．48％（P=0．013）;吸烟、不吸烟患者突变率分别为11．11％、39．29％（P=0．017）;外显子19突变8例（57．14％,8／14）,外显子21突变5例（35．71％,5／14）,外显子18、20同时突变1例（7．15％,1／14）。结论在重庆市地区NSCLC患者中,女性、非吸烟者EGFR突变率较高,EGFR基因突变以外显子19和21为主。     

非小细胞肺癌胸水中癌细胞EGFR基因TK区突变研究

目的:检测非小细胞肺癌(NSCLC)患者胸水标本中癌细胞EGFR基因TK区突变。方法:收集21例NSCLC患者胸水标本,经密度梯度离心后,将胸水中细胞包埋,制成蜡块;巢式聚合酶链式反应(PCR)扩增EGFR基因18-21号外显子;应用PCR-LIS-SSCP及直接测序方法检测EGFR基因TK区突变。结果:21例NSCLC患者胸水癌细胞标本,19例为腺癌,1例为腺鳞癌,1例为大细胞癌;9例患者存在EGFR基因TK区突变,均为腺癌,突变率为42.9%(9/21),腺癌患者突变率为47.4%(9/19);9例患者中8例接受TKIs治疗,疗效评价为PR(部分缓解)或SD(稳定)。5例出现TKIs耐药,疾病进展TTP(疾病进展时间)为4～17个月不等。应用TKIs治疗前后对比胸水及癌组织中EGFR基因突变,2例出现获得性突变。结论:NSCLC胸水标本与文献报道的癌组织中EGFR基因TK区突变率基本相同;胸水标本中检测肿瘤细胞基因突变的方法可靠,值得在临床推广、应用。     

肺腺癌EGFR基因19、21外显子突变与临床病理特征及预后的关系

目的分析肺腺癌表皮生长因子受体(EGFR)基因19、21外显子突变与临床病理特征及预后的关系。 方法回顾性分析2017年1月至2018年6月100例经病理证实为肺腺癌的患者临床资料与标本,采用PCR-ARMS技术检测标本EGFR基因19、21外显子突变情况,分析EGFR基因突变与临床病理特征及预后的关系。 结果100例肺腺癌标本共检测出47例EGFR突变,突变率为47.00%,其中第19号外显子突变20例(42.55%),包括6种形式的突变,以核苷酸框架缺失为主,最常见的类型为核苷酸从2234-2248位缺失15bp的delE746-A750突变,共11例,占55.00%;第21号外显子突变27例(57.45%),包括5种形式的突变,均是碱基置换突变,最常见的类型为2573位点的T被G取代的L858R,共17例,占62.96%;女性、无吸烟史、临床分期Ⅰ期患者EGFR19与EGFR21突变率高于其他患者(P<0.05);Logisitic回归分析显示性别、吸烟史、肿瘤直径、临床病理分期是影响EGFR19、EGFR21突变的因素。100例肺腺癌患者全部获得有效随访,EGFR19突变患者2年无进展生存率、总生存率分别为80.00%、85.00%,未突变患者分别为65.00%、73.75%;EGFR21突变患者2年无进展生存率、总生存率分别为81.48%、92.59%,未突变患者分别为63.01%、69.86%;女性、临床分期较早、无淋巴结转移、发生EGFR21突变患者2年生存率低于其他患者(P<0.05);Logisitic回归分析显示男性、临床分期较晚、有淋巴结转移、EGFR21未突变是肺腺癌患者不良预后的独立危险因素。 结论肺腺癌EGFR基因19、21外显子突变与性别、吸烟史、肿瘤直径、临床分期有关,同时也是预测预后的有效方法。     

非小细胞肺癌表皮生长因子受体突变的检测与分子靶向治疗

目的探讨非小细胞肺癌患者表皮生长因子受体(EGFR)基因突变率和突变类型,分析其临床特征,并观察EGFR突变与表皮生长因子受体酪氨酸激酶抑制剂(EGFR-TKIs)治疗疗效的相关性。方法收集203例非小细胞肺癌患者外科手术、淋巴结活检、经皮肺穿刺活检、气管镜活检和胸腔积液沉渣石蜡标本,应用ADx-ARMS法进行EGFR基因突变检测,分析基因的突变率及其与临床特征的关系;观察非小细胞肺癌(NSCLC)接受EGFR-TKIs治疗的非小细胞肺癌患者的疗效。应用SPSS23.0软件进行统计学分析,计数资料比较采用χ2检验,采用Kaplan-Meier法计算患者的PFS,采用Log-Rank检验分析各种因素对生存期的影响。结果 203例NSCLC患者,男性116例,女性87例,年龄为25~82岁。吸烟指数≥400支/者61例,小于400支/年和不吸烟者142例,腺癌152例,鳞癌21例,腺鳞癌14例,其他NSCLC16例。203例NSCLC患者EGFR总突变率为51.2%(104/203),包括19外显子缺失突变51例(49.0%),21外显子L858R突变44例(42.3%),19del及L858R总突变率占所有突变的96.1%,18外显子G719X点突变3例(2.9%),19del+L858R双突变3例(2.9%),1例20ins,2例T790M突变分别为1例19del+T790M和1例L858R+T790M。EGFR基因阳性突变率女性组高于男性组(66.7%vs.36.2%);非吸烟组高于吸烟组(63.4%vs.16.4%);腺癌组高于鳞癌组(53.3%vs.33.3%),P0.05。而EGFR基因突变状况与标本类型如手术、淋巴结活检、肺穿刺活检、气管镜活检和胸腔积液沉渣标本间无统计学差异,P=0.418。101例接受TKI治疗的NSCLC患者客观缓解率(ORR)为61.4%,疾病控制率(DCR)为71.3%,中位疾病无进展生存期(PFS)为10个月。其中EGFR突变阳性患者接受EGFR-TKIs治疗的ORR及DCR均要显著高于EGFR突变阴性及EGFR突变状态未明确人群(88.6%vs.16.7%vs.43.1%,P=0.000;95.5%vs.16.7%vs.56.9%,P=0.000)。EGFR突变阳性患者接受EGFR-TKIs治疗的中位PFS较EGFR突变阴性及EGFR突变状态未明确患者延长,有统计学差异(P=0.001)。进一步分析EGFR突变阳性19del组NSCLC患者ORR、DCR均高于L858R组(91.2%vs.85%,P=0.646;100%vs.90%,P=0.201);19del组NSCLC患者TKI治疗后中位PFS 14.5个月较L858R组10个月长,有统计学差异(P=0.010)。结论非小细胞肺癌患者EGFR突变高,以女性、不吸烟、腺癌为优势人群,EGFR敏感突变阳性者对EGFR-TKI疗效好,EGFR突变中19del者较L858R疗效更佳,基因检测结果可以较好地预测分子靶向药物的疗效,降低肿瘤进展的风险。     

老年非小细胞肺癌组织中表皮生长因子受体基因突变情况及其与切割修复交叉互补基因1表达的相关性

目的探讨老年非小细胞肺癌(NSCLC)组织中表皮生长因子受体(EGFR)基因突变情况及其与切割修复交叉互补基因(ERCC)1表达的关系。方法选择老年NSCLC患者150例作为研究对象,采用PCR技术检测老年NSCLC组织中EGFR基因突变和ERCC1基因表达水平。结果 150例老年NSCLC组织中EGFR基因突变62例,突变率为41.3%,外显子21和外显子19突变率分别为18.7%和18.0%。老年NSCLC组织中,男性EGFR基因突变率低于女性(P0.05),吸烟患者EGFR基因突变率低于非吸烟患者(P0.05),腺癌患者EGFR基因突变率高于非腺癌患者(P0.05),有肿瘤家族史患者EGFR基因突变率高于无肿瘤家族史患者(P0.05),不同TNM分期患者EGFR基因突变率比较有差异(P0.05)。EGFR基因突变型ERCC1高表达率低于EGFR基因野生型(P0.05)。结论老年NSCLC患者EGFR基因突变以外显子21和外显子19突变为主,性别、吸烟、腺癌、肿瘤家族史、TNM分期为EGFR基因突变的影响因素,EGFR基因突变者ERCC1基因表达量低。     

突变特异性免疫组织化学法检测非小细胞肺癌EGFR基因突变

目的探讨应用突变特异性免疫组织化学(IHC)法检测非小细胞肺癌(NSCLC)中表皮生长因子受体(EGFR)基因突变的临床使用价值。方法随机选取94例NSCLC患者,采用突变特异性IHC法和罗氏454下一代测序系统法进行检测EGFR基因突变状态,比较两种方法的特异性和灵敏度。结果 EGFR基因突变阳性定位在肿瘤细胞质和细胞膜或细胞膜中。突变特异性IHC法检测NSCLC患者中EGFR基因突变阳性率为36.17%,其中腺癌的突变率为48.44%,鳞癌的突变率为10.00%,差异有统计学意义(P<0.05),罗氏454下一代测序系统法检测NSCLC患者中EGFR基因突变阳性率为38.30%,其中腺癌的突变率为51.56%,鳞癌的突变率为10.00%,差异有统计学意义(P<0.05)。突变特异性IHC法能够有效检测NSCLC EGFR基因突变,其结果与罗氏454下一代测序系统法具有极好的一致性(P<0.001)。结论突变特异性IHC法能够有效检测NSCLC EGFR基因突变,并且突变特异性IHC法能够与EGFR酪氨酸激酶抑制剂(TKIs)治疗快速的反应,具有耗时短、易判读、检测费用少等特点。     

NSCLC患者呼出气冷凝液中p16基因突变的研究

目的 研究非小细胞肺癌患者(NSCLC)呼出气冷凝液(EBC)中p16基因突变的临床意义.方法 收集30例NSCLC患者和20例体检健康者的EBC标本,提取EBC中的DNA,对β-actin基因扩增阳性的EBC标本进行p16基因1、2、3号外显子PCR扩增,并进行DNA基因测序,用DNASTAR软件进行突变比对,结果进行统计学分析.结果 ①30例肺癌患者的EBC中,有26例β-actin基因片段扩增阳性,26例中有9例检出p16基因突变,突变率为34.6％.②9例肺癌患者发生p16基因突变的外显子,1号外显子3例,2号外显子5例,3号外显子1例.③26例肺癌患者EBC中β-actin基因片段扩增阳性中,Ⅰ期12例,p16基因突变3例,突变率25％;Ⅱ期7例,p16基因突变2例,突变率28.6％;Ⅲ期7例,p16基因突变4例,突变率57.1％.鳞癌14例,p16基因突变6例,突变率42.9％;腺癌11例,p16基因突变3例,突变率27.3％.结论 NSCLC患者EBC中可以检测到p16基因突变,特异性高,EBC中p16基因检测为肺癌研究提供新方法.     

EGFR基因突变与酪氨酸激酶抑制剂疗效及预后之间的关系

背景与目的表皮生长因子受体（epidermal growth factor receptor，EGFR）信号通路在非小细胞肺癌的发生和发展中起重要作用。EGFR酪氟酸激酶抑制剂（EGFR tyrosine kinase inhibitors，EGFR-TKIs）是目前非小细胞肺癌（NSCLC）治疗的热点。研究表明，EGFR基因突变与TKIs的疗效及预后相关。本研究旨在了解EGFR基因突变与两种酪氨酸激酶抑制剂疗效及预后的相关性。方法共收集了34例接受gefitinib单药治疗和25例接受erlotinib单药治疗的晚期NSCLC患者的病理组织蜡块及相关临床资料。用PCR-PAGE检测EGFR199b显子突变，PCR＊RFLP检测EGFR21外显子突变，均用直接测序进行验证。结合临床进行分析。结果59例NSCLC标本中共检测出22例标本中有EGFR基因突变，突变率为37．3％。EGFR基因突变率在女性、腺癌、不吸烟患者中高（P〈0．05）。有EGFR基因突变的患者接受酪氨酸激酶抑制剂治疗的有效率高于无突变患者（50％vs18．9％，P〈0．05），疾病控制率高于无突变患者（86．4％vs54．1％，P〈0．05）。有EGFR基因突变的患者的疾病进展时间和总生存期均高于无突变患者，但是没有统计学差异（P〉0.05）。结论EGFR基因突变在女性、腺癌和不吸烟者中发生率高。有EGFR基因突变的晚期NSCLC接受EGFR酪氨酸激酶抑制剂治疗的有效率和疾病控制率高于无EGFR基因突变者。     

老年非小细胞肺癌患者表皮生长因子受体基因19-21外显子突变研究

目的 研究老年非小细胞肺癌(NSCLC)患者表皮生长因子受体(epidermal growth factor receptor,EGFR)基因酪氨酸激酶区19-21外显子的突变状态,并对其临床特征进行初步分析.方法 选取46例病理确诊的老年非小细胞肺癌患者,提取肺癌组织或胸腔积液及心包积液肿瘤细胞基因组DNA,通过巢式PCR基因扩增和直接测序的方法 .分析19、20、21外显子的突变情况,并与其临床特征及应用酪氨酸激酶抑制剂(TKI)疗效的关系进行了初步分析. 结果 46例中,26例检测出带有基因突变(56.5%),其中非沉默突变者为19例(41.3%).19号外显子突变6例(13.0%),20号外显子突变13例(28.2%),21号外显子突变14例(30.4%).其中7例带有2处突变,其余均为单处突变.不吸烟者的突变发生率显著高于吸烟者(P＜0.01).应用酪氨酸激酶抑制剂(TKI)临床获益的患者带有EGFR 19、20、21外显子的基因突变的几率显著高于未获益者(P＜0.05).60～69岁与70～85岁年龄组基因突变状态差异无统计学意义. 结论 老年NSCLC患者EGFR基因酪氨酸激酶区19-21外显子突变特征与肺癌患者总体类似,与年龄关系不大,老年NSCLC患者同样可以通过基因检测获得TKI治疗预测信息.     

Epidermal growth factor receptor gene amplification and gefitinib sensitivity in patients with recurrent lung cancer

To evaluate the epidermal growth factor receptor (EGFR) protein expression, gene mutations and amplification as predictors of clinical outcome in patients with non-small-cell lung cancer (NSCLC) receiving gefitinib, we have performed fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC). We investigated the EGFR amplification and EGFR protein expression statuses in 27 surgically treated non-small-cell lung cancer (NSCLC) cases. These patients experienced relapse after surgery and received gefitinib 250 mg/day. The presence or absence of EGFR mutations of kinase domains was analyzed by genotyping analysis and sequences, and already reported. EGFR mutations were found from 15/27 lung cancer patients. EGFR mutation status was significantly correlated with better prognosis (log-rank test P = 0.0023). Smoking status (never smoker vs. smoker, P = 0.0032), and pathological subtypes (adenocarcinoma vs. non-adenocarcinoma, P = 0.0011), but not EGFR amplification (P = 0.1278), were correlated with survival of lung cancers. EGFR IHC results were correlated with FISH results (P = 0.0125), but not correlated with prognosis (P = 0.7921). Thus, the EGFR gene amplification or protein expression is not a predictor of gefitinib efficacy in Japanese patients with NSCLC. We have also evaluated the EGFR mutation status and clinico-pathological features for 27 NSCLC patients who had undergone surgery followed by treatment with gefitinib at the National Hospital Organization, Kinki-chuo Chest Medical Center. The EGFR mutation status, especially exon19 mutation was correlated with good response to gefitinib than exon 21 point mutation.     

Correlation of EGFR G873R mutation with prognosis of docetaxel in non-small cell lung cancer

BackgroundClinical features of epidermal growth factor receptor (EGFR) mutations have been commonly recognized in variant cancers. The role of EGFR mutations in non-small cell lung cancer (NSCLC) has spurred research and drug development efforts. However, there are still mutations that have not been widely reported, and their influences on NSCLC have not been fully elucidated; EGFR G873R mutation is just one of them. The aim of this study was to investigate the correlation between EGFR G873R mutation and the prognosis of chemotherapy in NSCLC.MethodsA total of 54 patients with NSCLC were enrolled in this study. Immunohistochemical staining was used to detect the expression of EGFR. A DNA extraction kit (GeneRead DNA FFPE Kit) was used to extract total DNA from resected cancer tissues. Genomic DNA targets were amplified by polymerase chain reaction (PCR), and then the amplicons were purified and sequenced. Statistical methods were performed to detect the relationship between EGFR G873R mutation and various clinicopathological features and the effect of EGFR G873R mutation on the prognosis of chemotherapy.ResultsEGFR G873R mutation did not show statistical significance, with EGFR high expression identified in 30 cases (P>0.05). Patients with EGFR G873R mutation had a significantly favorable prognosis of docetaxel (P=0.032), and for patients treated with docetaxel, EGFR G873R mutation was significantly correlated with better 5-year disease-free survival (DFS; P=0.026) and overall survival (OS; P=0.026). However, there was no statistical significance found between EGFR G873R mutation and the prognosis of vinorelbine (P>0.05), and for patients treated with vinorelbine, EGFR G873R mutation had no statistical significance with 5-year DFS (P>0.05) and OS (P>0.05).ConclusionsEGFR G873R mutation was remarkably correlated with the prognosis of docetaxel in NSCLC, which indicates that EGFR G873R may be employed as a promising biomarker to identify individuals with better prognosis of docetaxel and as an antitumor target for NSCLC treatment.     

Expression of LRIG1 is Associated With Good Prognosis for Human Non-small Cell Lung Cancer

Somatic mutations, which are associated with a certain rate of response to targeted therapies, are ubiquitously found in human non-small cell lung cancer (NSCLC). However, it is largely unknown which group of patients may benefit from the respective treatments targeting different somatic mutations. Therefore, more effective prognostic and predictive markers are desperately needed for the treatment of NSCLC harboring different somatic mutations. The leucine-rich repeats and immunoglobulin-like domains (LRIG)-1 is a tumor suppressor gene that belongs to the LRIG family. LRIG1 expression has prognostic significance in various human cancers.In this study, we first used the quantitative polymerase chain reaction (qPCR) and immunohistochemical analysis of 36 and 182 NSCLC patient tissues to analyze the LRIG1 expression respectively. To investigate the prognostic value of LRIG1 in NSCLC, we examined the correlation between clinical features and overall survival (OS) with Cox proportional hazard regression. We also compared the sensitivity and specificity of LRIG1 in NSCLC prognosis by logistic regression to further evaluate the prognostic efficiency of LRIG1 in NSCLC.We found that the LRIG1 expression was associated with pathological type, differentiation status, and stage of NSCLC. The result showed that LRIG1 was an independent prognostic factor for OS of NSCLC patients. LRIG1 in combination with other clinicopathological risk factors was a stronger prognostic model than clinicopathological risk factors alone.Thus, the LRIG1 expression potentially offered a significant clinical value in directing personal treatment for NSCLC patients.     

EGFR mutations and HER2/3 protein expression and clinical outcome in Chinese advanced non-small cell lung cancer patients treated with gefitinib

Background  To assess the role of various epidermal growth factor receptor (EGFR) mutations and HER2/3 protein expression as predictive markers of responsiveness to gefitinib therapy in Chinese patients with advanced non-small cell lung cancer (NSCLC). Methods  A total of 106 Chinese NSCLC patients who had failed at least one chemotherapy regimen received gefitinib 250 mg once daily. All the 106 tumors from these patients were screened for mutations in the EGFR exons 18–24, and 84 tumors were studied by immunohistochemistry for HER2/3 expression and correlated with clinical treatment outcome. Results  Patients with EGFR mutations had a significantly higher overall response rate (ORR), longer time to progression (TTP) and overall survival (OS) compared with those with wild-type receptor. No difference in ORR was observed between patients with exon 19 deletion and patients with other EGFR mutations. ORR in HER2-positive patients was significantly higher than in the HER2-negative group, irrespective of EGFR mutational status, and a trend for better ORR was observed for HER3-positive patients. HER2 and HER3 expression levels were not associated with any difference in terms of TTP and OS. Nevertheless, when considering the subgroups of non-responders to gefitinib, median TTP in patients with mutated EGFR was significantly longer than in those with no mutations (8.0 vs. 3.0 months, P = 0.0065). EGFR-mutated patients had no significant difference in ORR, TTP and OS according to HER2 and/or HER3 expression. Conclusions  EGFR mutations are effective predictors for gefitinib efficacy in Chinese patients with advanced NSCLC. HER2 and HER3 expression does not provide any additional information for selecting patients most likely to benefit from gefitinib treatment.