非小细胞肺癌中EML4-ALK融合基因的检测及其与EGFR、K-ras基因的相关性

目的 探讨不同类型非小细胞肺癌(NSCLC)的EML4-ALK融合基因的表达及其与表皮生长因子受体(EGFR)和K-ras基因突变的相关性.方法 用基因测序方法检测98例NSCLC患者肿瘤组织中EGFR(18、19、20、21外显子)基因和K-ras(12、13、61密码子)基因的突变.用荧光PCR法检测EML4-ALK融合基因的表达.用x2检验来分析EML4-ALK融合基因与EGFR、K-ras基因在NSCLC中的相关性.结果 98例样本中6例有EML4-ALK基因表达,表达率为6.1％,32例(32.7％)发生了EGFR基因突变,21例(21.4％)发生了K-ras基因突变.EML4-ALK融合基因阳性与患者年龄有关(x2=5.35,P=0.021),与性别(x2=0.48,P=0.491)、吸烟(x2=0.11,P=0.739)、组织学类型(x2=1.65,P=0.648)、腺癌分化程度(x2=1.59,P=0.452)无关.98例患者中,未发现有EGFR基因突变、K-ras基因突变和EML4-ALK融合基因表达同时存在的病例.结论 EML4-ALK融合基因代表了NSCLC的一个新的分子亚型,EML4-ALK融合基因与NSCLC患者年龄相关.EML4-ALK融合基因的表达与EGFR及K-ras基因突变不同时存在.     

经选择的肺腺癌患者中EML4-ALK融合基因发生率的研究

目的探讨表皮生长因子受体(EGFR)野生型肺腺癌患者EML4-ALK融合基因的发生率是否高于EGFR状态未明者。方法经病理确诊的不吸烟或少吸烟的肺腺癌患者100例,EGFR野生型组50例,EGFR状态未明组50例,采用原位荧光杂交法(FISH)检测EML4-ALK融合基因状态。结果EGFR野生型组15例(30%)ML4-ALK融合基因阳性;EGFR状态未明组4例(8%)EML4-ALK融合基因阳性,2组差异有显著性(P<0.05);EML4-ALK融合基因发生率与年龄、性别、临床分期无明显相关性(P>0.05)。结论在不吸烟或少吸烟的肺腺癌患者中,经EGFR基因状态筛选可提高EML4-ALK融合基因检测阳性率。     

EML4-ALK融合基因在非小细胞肺癌中的研究

<正>肺癌全球每年新增病历约160万,其中非小细胞肺癌(NSCLC)占80%~85%[1]。EML4-ALK在肺癌中的阳性率大致约7%[2],与表皮生长因子受体(EGFR)、KRAS基因突变很少同时存在。针对这一NSCLC新的靶点,本文就EML4-ALK及其特征、致病机制、检测方法,以及其临床特征和治疗等综述如下。EML4、ALK、EML4-ALK简介EML4位于人类染色体2p21,由三部分组成:氨基酸末端碱基区、疏水的棘皮动物微管相关蛋白区、WD重复区;ALK位于人类染色体2p23,含29个外显子,全长728kb,具有RTK三个部分:跨膜区、胞外     

EGFR突变状态与肺腺癌新分类中组织学亚型和生存关系

目的本研究选取手术切除的中晚期肺腺癌中常见病理亚型,通过检测EGFR突变状态探讨其对患者生存时间的影响。方法收集我院胸外科经手术切除的Ⅱa-Ⅲa期肺腺癌患者生存资料,通过检测肺腺癌EGFR基因,比较EGFR基因突变阳性与EGFR突变基因阴性(野生型)患者的2年无疾病生存率(DFS)和5年总生存率(OS),分析比较EGFR突变状态对患者生存的影响。结果在120例肺腺癌患者中EGFR基因突变率为46.6%,最常见的突变位点是外显子19(44.6%)和外显子21(42.8%),在所有浸润性肺腺癌组织学类型中,最常见的组织学类型是腺泡为主型(55.8%),其次是伏壁为主型(25.8%),两种组织学类型中的EGFR突变阳性率为(腺泡型61.2%VS伏壁为主型44.7%),EGFR野生型(腺泡型55.3%VS伏壁型33.8%),两者组织学亚型中EGFR突变率差异无统计学意义(P=0.192)。通过比较生存时间分析得出,肺腺癌2年无疾病生存率(EGFR突变阳性55.3%VS EGFR突变野生型55.6%,P=0.367),5年总生存率(EGFR突变阳性55.5%VS EGFR突变野生型40.6%,P=0.143),差异均无统计学意义。结论在手术切除的中晚期肺腺癌患者中,EGFR突变状态本身不是影响肺腺癌术后生存时间的因素。     

实时荧光定量PCR法检测肺腺癌患者恶性胸腔积液中EMIA—ALK融合基因突变的研究

目的：探讨应用实时荧光定量PCR（ FQ-PCR）方法检测原发性肺腺癌患者恶性胸腔积液中的EML4-ALK融合基因突变情况,并对指导克唑替尼治疗的可行性进行探讨。方法收集39例原发性肺腺癌患者的胸腔积液,使用FQ-PCR法检测EML4-ALK融合基因的阳性率。结果胸腔积液中EML4-ALK融合基因的阳性率为12.8%,克唑替尼治疗对于EML4-ALK融合基因的和无EML4-ALK融合基因的患者相比有较高的疾病控制率（80% vs 16.7%,P〈0.05）。结论对难以获得组织标本的肺癌患者可选用FQ-PCR法检测肺癌患者恶性胸腔积液中EML4-ALK以指导克唑替尼的应用。     

棘皮动物微管结合蛋白4-间变淋巴瘤激酶融合基因与非小细胞肺癌的关系

近年来,针对表皮生长因子受体(epidermal growth factor receptor,EGFR)的酪氨酸激酶抑制剂(tyrosine kinase inhibitors,TKI)已成为非小细胞肺癌(non-small cell lung cancer,NSCLC)治疗方面的里程碑性进展,但对EGFR突变阴性者疗效欠佳,且部分病例存在获得性耐药,因此,寻找新的分子靶点成为NSCLC治疗中的关键问题.棘皮动物微管结合蛋白4-间变淋巴瘤激酶(anaplastic lymphoma kinase with the echinoderm microtubule-associated protein-like 4,EML4-ALK)是在肺癌患者中发现的新型融合基因,其与EGFR及v-Ki-ras2大鼠Kirsten肉瘤病毒癌基因同源基因(v-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog,KRAS)突变不共存,且含有该融合基因的肺癌患者具有特定的临床特征,提示该基因是肺癌特异性较高的分子标志物.除此之外,Ⅱ期临床试验结果表明,ALK-TKI对EML4-ALK阳性的NSCLC患者有显著的抗肿瘤作用.基于以上两点,EML4-ALK可能成为继EGFR之后的又一分子靶点,其分子特点、临床特征、检测方法及临床试验等多方面均引起了人们的广泛关注.现将EML4-ALK融合基因的有关内容综述如下.     

实时荧光定量PCR法检测肺腺癌患者恶性胸腔积液中EML4-ALK融合基因突变的研究

目的探讨应用实时荧光定量PCR(FQ-PCR)方法检测原发性肺腺癌患者恶性胸腔积液中的EML4-ALK融合基因突变情况,并对指导克唑替尼治疗的可行性进行探讨。方法收集39例原发性肺腺癌患者的胸腔积液,使用FQ-PCR法检测EML4-ALK融合基因的阳性率。结果胸腔积液中EML4-ALK融合基因的阳性率为12.8%,克唑替尼治疗对于EML4-ALK融合基因的和无EML4-ALK融合基因的患者相比有较高的疾病控制率(80%vs 16.7%,P0.05)。结论对难以获得组织标本的肺癌患者可选用FQ-PCR法检测肺癌患者恶性胸腔积液中EML4-ALK以指导克唑替尼的应用。     

非小细胞肺癌靶向治疗的一个新趋势:EML4-ALK融合基因

在非小细胞肺癌个体化治疗成为共识的今天,靶向治疗是其中的重要组成部分,EML4-ALK是近期发现的非小细胞肺癌的新的分子亚型.在亚洲、女性、无或轻度吸烟、肺腺癌患者中的高检出率及ALK抑制剂PF-02341066在Ⅰ期临床试验中显示出的良好疗效使EML4-ALK成为非小细胞肺癌又一靶向治疗点.     

非小细胞肺癌患者EGFR、ALK基因突变及临床病理特征与预后的相关性研究

目的探讨非小细胞肺癌(NSCLC)患者EGFR、ALK基因突变状态及病理特征,分析二者与患者预后情况相关性。方法收集2015年1月至2018年1月苏州大学附属第一医院病理确诊的NSCLC病例262例,记录患者临床病理特征及转归,采用突变增阻滞系统(ARMS)-Taqman探针法及RT-PCR检测患者肿瘤样本的EGFR和ALK基因外显子突变情况,采用Kaplan-Meier法和Cox风险回归考察不同基因突变情况的预后、临床病理特征与临床结局的相关性。结果262例NSCLC患者中,168例为野生型,ALK突变12例,突变率4.6%;EGFR突变82例,突变率31.30%,其中18号外显子突变2例,19号外显子突变37例,20号外显子突变9例,21号外显子突变31例;各组间年龄、病理类型比较差异有统计学意义,P<0.05;Wild type组中位OS时间11.6月,ALK组中位OS时间15.0月,EGFR组中位OS时间36.8月,EGFR组OS明显优于ALK组OS时间及Wild type组OS时间,P<0.05;EGFR19号外显子突变组、21号外显子突变组OS生存时间显著高于18号外显子突变组和20号外显子突变组,P<0.05;EGFR阴性(RR=15.751,95%CI:9.252~26.816)、鳞癌(RR=18.344,95%CI:6.187~54.388)为OS的危险因素。结论EGFR突变状态、病理类型对非小细胞肺癌预后具有良好的预测价值,鳞癌和EGFR阴性患者生存期短。EGFR基因少见突变(18号外显子、20号外显子)患者的中位OS时间更短。     

肺鳞癌分子靶向治疗的前景柳暗花明

以表皮生长因子受体酪氨酸激酶抑制剂( epidermal growth factor receptor tyrosine kinase inhibitor,EGFR-TKI)的临床研究与应用为契机,进展期非小细胞肺癌( non-small-cell lung cancer,NSCLC)的治疗已进入个体化和靶向治疗时代,其中以EGFR-TKI及棘皮动物微管结合蛋白4-间变淋巴瘤激酶(EML4-ALK)融合基因抑制剂克唑替尼(crizotinib)最为振奋人心,两类药物靶点明确、疗效确切,可显著改善EGFR敏感突变及EML4-ALK融合患者的生存.     

癌胚抗原在不同驱动基因肺癌患者中的差异性

目的 通过回顾性分析不同驱动基因肺癌患者的血清癌胚抗原(CEA)水平,探讨CEA在不同驱动基因肺癌患者中的差异性.方法 选择2015年9月至2018年12月唐都医院呼吸与危重症医学科经治的210例肺癌患者,分别采用电化学发光免疫分析法和探针扩增阻滞突变系统聚合酶链反应法检测CEA水平及基因突变状况,分析不同驱动基因肺癌...     

非小细胞肺癌中EML4－ALK融合基因的检测及分析

目的检测非小细胞肺癌（NSCLC）患者中EML4-ALK基因的表达率并分析其与患者的临床病理学特征的关系。方法用荧光PCR法检测我院98例NSCLC患者肿瘤组织中EML4-ALK基因的表达。结果98例NSCLC患者中检测出6例有EML4-ALK基因的表达,表达率为6．12％。6例阳性患者中,4例为女性,2例为男性;不吸烟者4例,吸烟者2例;年龄〈50岁者5例,≥50岁者1例;5例为腺癌患者,1例为腺鳞癌患者;5例为中分化患者,1例为低分化患者,高分化患者中未发现阳性。结论EML4-ALK融合基因代表了NSCLC的-个新的分子亚型,表达EML4-ALK融合基因的NSCLC具有其独特的临床病理学特点。     

High prevalence of gene abnormalities in young patients with lung cancer

Background

Recently, driver oncogenes in adenocarcinoma of the lung were identified, and several molecular target agents were introduced in the clinical setting. However, there are few reports on the frequency of gene abnormalities in young patients with lung cancer.

Materials and methods

Twelve patients with lung adenocarcinoma aged 40 or younger at Juntendo University Urayasu Hospital or Juntendo University Hospital from July 2004 to March 2010 were analyzed for driver oncogene status including EGFR activating mutation, EML4-ALK fusion gene, and K-ras mutation.

Results

Four patients showed EGFR gene mutation. Five out of 7 EGFR mutation-negative patients showed positive results for EML4-ALK gene fusion. One case whose EGFR mutation was indeterminate.

Conclusions

Driver oncogene including EGFR mutation and EML4-ALK fusion gene was identified in 9 of 12 cases (75%). Examination of gene abnormalities is essential in young patients with non-small cell lung cancer to provide the best treatment.KEY WORDS : Young patients, driver oncogene, lung cancer, EGFR, EML4-ALK     

Therapeutic strategies to overcome crizotinib resistance in non-small cell lung cancers harboring the fusion oncogene EML4-ALK

The echinoderm microtubule-associated protein-like 4 (EML4)-anaplastic lymphoma kinase (ALK) fusion oncogene represents a molecular target in a small subset of non-small cell lung cancers (NSCLCs). This fusion leads to constitutive ALK activation with potent transforming activity. In a pivotal phase 1 clinical trial, the ALK tyrosine kinase inhibitor (TKI) crizotinib (PF-02341066) demonstrated impressive antitumor activity in the majority of patients with NSCLC harboring ALK fusions. However, despite these remarkable initial responses, cancers eventually develop resistance to crizotinib, usually within 1 y, thereby limiting the potential clinical benefit. To determine how cancers acquire resistance to ALK inhibitors, we established a model of acquired resistance to crizotinib by exposing a highly sensitive EML4-ALK-positive NSCLC cell line to increasing doses of crizotinib until resistance emerged. We found that cells resistant to intermediate doses of crizotinib developed amplification of the EML4-ALK gene. Cells resistant to higher doses (1 μM) also developed a gatekeeper mutation, L1196M, within the kinase domain, rendering EML4-ALK insensitive to crizotinib. This gatekeeper mutation was readily detected using a unique and highly sensitive allele-specific PCR assay. Although crizotinib was ineffectual against EML4-ALK harboring the gatekeeper mutation, we observed that two structurally different ALK inhibitors, NVP-TAE684 and AP26113, were highly active against the resistant cancer cells in vitro and in vivo. Furthermore, these resistant cells remained highly sensitive to the Hsp90 inhibitor 17-AAG. Thus, we have developed a model of acquired resistance to ALK inhibitors and have shown that second-generation ALK TKIs or Hsp90 inhibitors are effective in treating crizotinib-resistant tumors harboring secondary gatekeeper mutations.     

High Incidence of EGFR Mutations in Pneumonic-Type Non-Small Cell Lung Cancer

To retrospectively identify computed tomography (CT) features that correlate with epidermal growth factor receptor (EGFR) mutation in surgically resected pneumonic-type lung cancer (P-LC).A total of 953 consecutive patients with surgically resected lung cancer in the First Affiliated Hospital of Guangzhou Medical University from August 2011 to August 2013 were studied. The CT manifestations were reevaluated independently by 2 radiologists. The presence of pneumonic-type consolidation with pathological confirmed non-small lung cancer (NSCLC) was defined as P-LC. EGFR mutation was determined by direct DNA sequencing or amplification refractory mutation system-PCR. EGFR mutation rates as well as clinical and pathological manifestations between P-LC and control lung cancer patients were compared.P-LC was diagnosed in 85 patients. Among these patients, 82 were adenocarcinoma (including 78 cases of invasive adenocarcinoma and 4 cases of microinvasive adenocarcinoma), 2 were squamous carcinoma and 1 was other type. P-LC occurred more frequently in female (58.8% vs 37.1%, P < 0.01), nonsmoking (76.5% vs 56.5%, P = 0.001) and adenocarcinoma (58.8% vs 37.1%, P < 0.01) patients. Moreover, EGFR mutations were found in 39 of 52 P-LC patients (75%) and 263 of 542 non-P-LC NSCLC patients (48.5%). However, no difference was found on the mutation sites of EGFR. Histological type, sex, and radiological manifestations (P-LC vs non-P-LC) but not smoking or sequencing method can be served as the independent predictor of EGFR mutations.P-LC patients showed a significant higher incidence of EGFR mutations, which was independent of sex, histological type, and smoking history. The patients with imaging manifestation of pneumonic-type consolidation are highly suggested to perform EGFR mutation analysis to guide the sequential treatment.     

肺腺癌中PD-L1、B7H3的表达与EGFR基因突变的关系

目的探讨程序性凋亡配体1(programmed death-ligand 1,PD-L1)、B7H3在肺腺癌中的表达与表皮生长因子受体(epidermal growth factor receptor,EGFR)基因突变状况的关系,为EGFR基因突变的肺腺癌患者提供一种新的临床治疗方法。方法收集78例手术切除的肺腺癌组织,所有标本均有EGFR基因检测结果。应用免疫组织化学PV-6 000法检测PD-L1、B7H3的表达情况。结果肺腺癌组织中有32例(41.0%)发生EGFR突变,其中女性的突变率高于男性(30.8%vs 10.3%,P0.05),不吸烟者比吸烟者有更高的突变率(28.2%vs12.8%,P0.05)。PD-L1、B7H3在EGFR突变型患者中的阳性率分别为71.9%、68.8%,均高于其在野生型患者中的发生率45.7%、43.5%(P0.05),且均与21外显子突变密切相关(P0.05),但未发现与19外显子突变相关(P0.05)。PD-L1在女性、不吸烟者及II~III期中的表达水平高于男性、吸烟者及I期患者(P0.05)。B7H3的表达与TNM分期及淋巴结转移情况相关(P0.05)。PD-L1与B7H3表达呈正相关关系(P0.05)。结论 PD-L1、B7H3高表达在EGFR突变型肺腺癌的发生发展中起重要作用,以PD-L1、B7H3为靶点的免疫治疗,可能成为这部分患者治疗的新方法。