非小细胞肺癌患者恶性胸腔积液中EGFR基因突变

非小细胞肺癌患者接受表皮生长因子受体-酪氨酸激酶抑制剂(EGFR-TKI)治疗前进行EGFR检测已经成为共识.恶性胸腔积液是非小细胞肺癌(NSCLC)患者常见的临床表现,采用不同方法对胸腔积液细胞及游离核酸提取后进行EGFR检测,对预测EGFR-TKI的疗效有着良好前景.     

非小细胞肺癌表皮生长因子受体基因突变的异质性

表皮生长因子受体(EGFR)酪氨酸激酶抑制剂(TKI)在EGFR基因活化突变阳性晚期非小细胞肺癌(NSCLC)患者中疗效显著,然而,同样是EGFR活化突变阳性的NSCLC患者接受EGFR-TKI治疗后,疗效仍有明显差异;同一个患者的不同瘤灶也会对EGFR-TKI表现出不同的反应,这些现象可能与EGFR基因突变异质性有关,同一瘤灶的不同部分、同一患者的不同瘤灶、同一瘤灶治疗前后,EGFR突变状态都有可能不一致.文章从三个方面对EGFR突变异质性研究进展作一介绍.     

非小细胞肺癌胸腔积液与配对肿瘤组织中EGFR基因突变检测的比较

目的:探讨非小细胞肺癌胸腔积液与配对肿瘤组织标本中EGFR基因突变检测结果的一致性,评价胸腔积液标本检测EGFR基因突变的应用价值.方法:收集非小细胞肺癌患者胸腔积液与配对肿瘤组织样本72例,采用ARMS方法,检测样本中EGFR基因第18~21外显子突变情况.结果:细胞学样本和组织学样本中EGFR基因突变阳性率分别为48.61%和51.39%,两者差异无统计学意义(P>0.05),二者一致率为92.11%,不一致率为7.89%.结论:二者的一致率较高,恶性胸水可以作为无法获得肿瘤组织的晚期非小细胞肺癌EGFR基因检测的有效样本.     

洛铂胸腔灌注治疗晚期非小细胞肺癌患者恶性胸腔积液的临床观察

目的观察洛铂胸腔内灌注化疗治疗晚期非小细胞肺癌（NSCLC）恶性胸腔积液的近期疗效和患者不良反应。方法回顾性分析25例晚期NSCLC恶性胸腔积液患者（A组）,胸腔置管引流后腔内灌注洛铂和白细胞介素2,以106例胸腔内灌注顺铂的患者（B组）为对照。比较两组近期疗效、生命质量和不良反应。结果两组均可在短期内较好控制恶性胸腔积液,疾病控制率分别为88．0％（22／25）和81．1％（86／106）,两组差异无统计学意义（P〉0．05）。A组生命质量控制率为84．0％（21／25）,B组为64．2％（68／106）,两组差异有统计学意义（P=0．0381）。患者不良反应主要为消化道和血液学反应,其中消化道反应两组差异有统计学意义（P=0．0349）。结论洛铂腔内化疗治疗晚期NSCLC患者恶性胸腔积液的近期疗效较好,能改善患者生命质量,且不良反应较轻,值得临床推广应用。     

非小细胞肺癌EGFR基因突变的临床意义研究

  目的   探讨非小细胞肺癌EGFR基因外显子突变与其临床病理特征的关系。   方法   利用ADx-ARMS?EGFR基因突变检测试剂盒,检测214例未接受过Gefitinib治疗的非小细胞肺癌患者组织中EGFR基因外显子18、19、20和21突变。   结果   非小细胞肺癌组织中EGFR基因总突变率为45.8%（98/214）,外显子18、19、20和21的突变率分别为0.93%（2/214）、22.0%（47/214）、2.3%（5/ 214）和20.6%（44/214）。另有2例19和21外显子双重突变。EGFR基因在肺腺癌组织中的总突变率为50.3%（93/185）明显高于肺鳞状细胞癌17.2%（5/29）（P=0.001）。EGFR基因在女性患者中的突变率57.0%（57/100）高于男性36.0%（41/114）（P=0.002）,EGFR基因在NSCLC淋巴结转移患者中的突变率（66.7%）显著高于无淋巴结转移患者（39.5%）（P < 0.05）,但EGFR基因突变率与肺癌患者的年龄、肿瘤分级和临床分期均无显著性差异（P>0.05）。   结论   中国肺癌尤其是肺腺癌患者存在EGFR基因的较高突变率,EGFR外显子19、21突变结合肺癌的临床病理特征有望成为评估TKI治疗非小细胞肺癌疗效的分子标志。      

EGFR mutations predict a favorable outcome for malignant pleural effusion of lung adenocarcinoma with Tarceva therapy

The aim of the present study was to evaluate the therapeutic effects and adverse reactions of Tarceva treatment for malignant pleural effusion (MPE) caused by metastatic lung adenocarcinomas. One hundred and twenty-eight patients who failed first-line chemotherapy drug treatment were divided into a mutation and a non-mutation group according to the presence or absence of epidermal growth factor receptor (EGFR) mutations. Each patient received closed drainage combined with simple negative pressure suction after thoracoscopic talc poudrage pleurodesis and oral Tarceva treatment. Short-term and long-term clinical therapeutic effects of Tarceva were evaluated. The EGFR mutation rate in pleural metastatic tissues of lung adenocarcinoma acquired through video-assisted thoracoscopic surgery was higher compared to that in surgical resection specimens, plasma specimens and pleural effusion specimens compared to previously reported results. There were significant statistical differences in the average extubation time (p<0.01), drainage volume of pleural effusion (p<0.05), Karnofsky score and formation of encapsulated pleural effusion 4 weeks after surgery (p<0.05) between these two groups. The number of patients with mild pleural hypertrophy in the mutation group was significantly higher compared to the non-mutation group (p<0.01), while the number of patients with severe pleural hypertrophy was significantly reduced (p<0.05). There was significant statistical discrepancy between these two groups in terms of improvement of peripheral blood carcinoembryonic antigen and tissue polypeptide antigen after 4 weeks of therapy. The complete remission rate and the efficacy rate were higher in the mutation group compared to that in the non-mutation group (p<0.05). There was a longer overall survival time after Tarceva treatment in patients with EGFR mutations than those without EGFR mutation. EGFR mutations predict a favorable outcome for malignant pleural effusion of lung adenocarcinoma with Tarceva therapy. Detection of EGFR mutations may determine the responsiveness of malignant pleural effusion to Tarceva treatment.     

Detection and comparison of epidermal growth factor receptor mutations in cells and fluid of malignant pleural effusion in non-small cell lung cancer

Cells or cell-free fluid of malignant pleural effusion could be important clinical specimen for epidermal growth factor receptor (EGFR) mutation screening in advanced non-small cell lung cancer (NSCLC) patients. However, their usefulness in mutation detection has not been well compared. In this study we recruited 26 East Asian NSCLC patients with malignant pleural effusion, determined the mutation status of EGFR in both cells and matched cell-free fluid with the use of sequencing and mutant-enriched PCR. After comparing the mutation spectrums, we found both the cells and cell-free pleural fluid may be feasible clinical specimen for EGFR mutation detection in unresectable NSCLC given sensitive genotyping assays employed. Direct sequencing could miss a significant portion of mutations in these heterogeneous specimens. More sensitive methods, such as mutant-enriched PCR and gene scan, could provide more reliable mutational information.     

EGFR and KRAS mutations in lung carcinomas in the Dutch population: increased EGFR mutation frequency in malignant pleural effusion of lung adenocarcinoma

Background

Frequencies of EGFR and KRAS mutations in non-small cell lung cancer (NSCLC) have predominantly been determined in East Asian and North American populations, showing large differences between these populations. The aim of the present study was to determine the frequency of EGFR and KRAS mutations in NSCLC in the West European Dutch population in primary carcinomas and different metastatic locations.

Methods

EGFR (exons 19, 20 and 21) and KRAS (exons 2 and 3) mutation test results of NSCLC samples of patients in 13 hospitals were collected. The tests were performed on paraffin-embedded tissue or cytological material of primary and metastatic lung carcinomas.

Results

EGFR mutations were detected in 71/778 (9.1?%) tested patients; in 66/620 (10.6?%) adenocarcinomas. EGFR mutations were significantly more often detected in female than in male patients (13.4?% vs. 5.5?%, p?<?0.001). KRAS mutations were found in 277 out of 832 (33.3?%) tested patients; in 244/662 (36.9?%) adenocarcinomas. A significantly increased frequency of EGFR mutations was observed in patients with malignant pleural/pericardial effusions (26.5?%; odds ratio (OR) 2.80, 95?% confidence interval (CI) 1.22?C6.41), whereas the frequency of KRAS mutations was significantly decreased (18.8?%; OR 0.35, 95?% CI 0.14?C0.86).

Conclusions

In the investigated Dutch cohort, patients with malignant pleural/pericardial effusion of lung adenocarcinoma have an increased frequency of EGFR mutations. The overall frequency of EGFR mutations in lung adenocarcinomas in this West European population is within the frequency range of North American and South European populations, whereas KRAS mutation frequency is higher than in any population described to date.     

ATP生物荧光肿瘤体外药敏检测技术指导复发合并恶性胸腔积液的非小细胞肺癌的治疗

Objective: This study was aimed to research the feasibility of ATP-bioluminescence assay (ATP-TCA) guiding the treatment on recurrent non-small cell lung cancer (NSCLC) combined with malignant pleural effusion. Methods: We collected 30 pleural fluid samples which were approved to be positive by cytology from recurrent NSCLC patients. These cells were cocultured with chemotherapy medicines, single agent or drugs combination. Five drug concentrations, two parallel holes were examined in vitro for 4 days, the results were measured by adding luciferase-fluorescein working system and luminescence analyzer. We applied chemotherapy medicines according to the results in vitro of ATP-TCA. Results: There were differences among drug sensitivities of individuals. All the samples could be evaluated. Effective single drugs included cisplatinum, mitomycin C, doxorubicin, and pemetrexed disodium; sensitive drugs in the combination therapy were gemcitabine plus cisplatin, vinorelbine plus cisplatin, paclitaxel plus cisplatin, docetaxel plus cisplatin, and mitomycin C, vindesine plus cisplatin, in which gemcitabine + cispiatin (GEM + DDP) in vitro was the most efficient program. Conclusion: ATP-TCA in vitro sensitivity assay is rapid, reliable, and simple to guide the treatment of recurrent NSCLC with malignant pleural effusion, and can help clinicians to make the individual chemotherapy program.     

综合治疗非小细胞肺癌并恶性胸水疗效观察

目的 探讨非小细胞肺癌并恶性胸水的综合治疗方法.方法 采用博来霉素加力尔凡胸腔灌注并原发灶及纵隔转移之淋巴结外照射.2001年2月至2002年2月治疗非小细胞肺癌并恶性胸水30例,均由病理证实,采用DF方案化疗两个周期后行博来霉素加力尔凡胸腔灌注.其中腺癌8例,鳞癌 15例,鳞腺癌7例.生活质量水平评分60～70分14例,50～60分16例.结果 胸水全部消失21例,胸水残留500 ml以下3例,残留1000 ml以下6例.原发灶CR 21例,PR 5例,SD 3例,PD 1例.坚持3年随访,其中21例均存活36个月并无胸水复发及远地转移,6例死于全身多脏器转移.3例骨髓抑制,白细胞3.0×109/L左右,不时靠重组人粒巨细胞集落刺激因子或口服中成药维持.结论 非小细胞肺癌DF方案化疗两周期后博来霉素加力尔凡胸腔灌注后并外照射是综合治疗非小细胞肺癌合并恶性胸水的很好治疗方法.     

贝伐单抗联合化疗在非小细胞肺癌恶性胸腔积液患者中的应用效果

目的探讨贝伐单抗联合化疗在治疗非小细胞肺癌合并恶性胸腔积液患者中的临床应用效果。方法选取2013年1月至2016年6月间青岛市中心医院收治的86例非小细胞肺癌合并恶性胸腔积液患者为研究对象,采用随机数表法分成两组,每组43例,其中,观察组患者在常规化疗基础上添加贝伐单抗联合治疗,对照组患者接受常规化疗。比较两组患者的治疗效果,同时对患者治疗过程中发现的蛋白尿、高血压、发热、疼痛和脱发等不良反应情况进行统计,并将不良反应按照从大到小严重程度分为Ⅰ级、Ⅱ级、Ⅲ级和Ⅳ级不良反应。结果观察组患者总有效率为74.4%,较对照组患者总有效率51.2%高,差异有统计学意义(P<0.05)。观察组Ⅳ级不良反应发生率少于对照组,差异有统计学意义(P<0.05)。结论贝伐单抗联合化疗治疗非小细胞肺癌合并恶性胸腔积液患者有显著效果,能有效减少恶性胸腔积液的产生,较大程度减少患者不良反应的发生情况,适合临床推广。     

Application of non-small cell lung cancer pleural effusion cell blocks in molecular pathological detection

Objective:The tumor tissues used in molecular pathological detection were usual y obtained by surgery, which would cause trauma and may not be suitable for the terminal cancer patients. This paper evaluated the value of the non-smal celllung cancer (NSCLC) pleural ef usion cellblocks as tumor tissues replacement materials in the application of molecular pathological detection. Methods: Tumor cells were made into cellblocks through stratified centrifugal from 30 NSCLC pa-tients with the pleural ef usion. The immunohistochemistry, fluorescence in situ hybridization (FISH) and gene sequencing methods were employed in our experiments. Results:The tumor cells of cellblock section were rich and could keep part of histological structure. Immunohistochemistry staining could assist diagnosis and tumor parting. Epidermal growth factor receptor (EGFR) FISH-positive was found in 33.33%of the group, high polysomy in 6 cases, amplification in 4 cases. EGFR gene mutations were found in 8 cases of 30 samples, with an incidence of 26.67%, 6 cases were detected in the exon 19, and 2 cases were detected in the exon 21. Conclusion:The NSCLC pleural ef usion cellblocks are useful for the diagnosis and determining the primary source of tumor, instructed targeted therapy.     

176例非小细胞肺癌的EGFR基因突变分析

目的 分析非小细胞肺癌（NSCLC）中上皮生长因子受体（EGFR）基因突变的发生率和突变类型。方法 收集123例正常肺组织和176例肺癌组织,采用PCR扩增和基因测序方法,对组织DNA中EGFR外显子19～21基因突变进行分析。结果 正常肺组织中EGFR基因均为野生型,肺癌组织中EGFR基因突变检测率为32．4％（57／176例）,其中,外显子19和21突变分别占突变总数的64．9％（37／57例）和31．6％（18／57例）,外显子20突变少见,仅占3．5％（2／57例）。外显子19突变发生在第746～753位密码子,均为碱基缺失突变,有7种不同类型。外显子20突变发生在第789—793位密码子,为碱基替换突变。外显子21突变全部是第858位密码子碱基替换突变。EGFR基因突变多见于女性,肺腺癌和腺鳞癌。结论 EGFR基因突变是一种肿瘤特异性的体细胞遗传改变,突变发生率约占肺癌总数的1／3,其中以外显子19和21为主。女性、肺腺癌和腺鳞癌中突变多见。     

High sensitivity detection of epidermal growth factor receptor mutations in the pleural effusion of non-small cell lung cancer patients

Epidermal growth factor receptor (EGFR) mutations are a strong determinant of tumor response to gefitinib in non-small cell lung cancer (NSCLC). We attempted to elucidate the feasibility of EGFR mutation detection in cells of pleural effusion fluid. We obtained 24 samples of pleural effusion fluid from NSCLC patients. The pleural effusion fluid was centrifuged, and the cellular components obtained were used for detection. EGFR mutation status was determined by a direct sequencing method (exons 18-21) and by the Scorpion Amplified Refractory Mutation System (ARMS) method. EGFR mutations were detected in eight cases. Three mutations were detected by both methods, and the other five mutations were detected by Scorpion ARMS alone. The mutations were detected by both methods in all four partial responders among the seven patients who received gefitinib therapy. Direct sequencing detected the mutations in only two of four cases with partial response. These results suggest that the DNA in pleural effusion fluid can be used to detect EGFR mutations. The Scorpion ARMS method appears to be more sensitive for detecting EGFR mutations than the direct sequencing method.     

伴恶性胸腔积液Ⅳ期NSCLC原发肿瘤三维放疗的临床分析

目的 回顾性探讨伴恶性胸腔积液Ⅳ期NSCLC原发灶三维放疗的有效性和安全性。方法 选择2007-2018年经病理诊断、初诊的伴恶性胸腔积液Ⅳ期NSCLC198例,按治疗状况分为未治疗组45例、药物治疗组57例、放疗组96例。分析放疗组和药物组近期疗效、总生存和不良反应。Kaplan-Meier法计算生存率并log-rank检验,Cox模型多因素预后分析。结果 放疗组有效率为54%、无效率为46%,显著优于药物组的25%、75%(P=0.007);1、2、3、5年总生存和中位生存期放疗组分别为47%、18%、6%、1%和12个月,显著高于药物组的15%、3%、2%、0%和5个月(P<0.001)。多因素分析显示增加原发灶放疗是延长生存的影响因素(P<0.001), ≥63Gy、4~6周期化疗有延长生存趋势(P=0.063、0.071),EGFR敏感突变者放疗联合分子靶向治疗总生存显著优于联合化疗而EGFR突变未知者(P=0.007)。增加原发灶放疗未增加明显的不良反应(P>0.05)。结论 伴恶性胸腔积液Ⅳ期NSCLC原发灶三维放疗可能延长患者总生存,增加三维放疗的不良反应可耐受。     

The prognostic significance of malignant pleural effusion at the time of thoracotomy in patients with non-small cell lung cancer

Surgery is usually not indicated for malignant pleural effusion (PE) due to its poor prognosis. However, PE is first detected at thoracotomy, and it is difficult to judge an appropriate mode of resection. Forty-nine patients with lung cancer were first diagnosed as PE and/or pleural dissemination (PD) at thoracotomy. The histological types were 36 adenocarcinoma, ten squamous cell carcinoma and three large cell carcinoma. Sixteen patients had only PE, 17 had only PD, and 16 had both PE and PD. Ten patients underwent only exploratory thoracotomy, seven partial resection, 27 lobectomy and five panpleuropneumonectomy. The overall survival rate was 26.7% at 3 years. The patients with PE and/or PD seemed to have a poorer survival compared to our previous study. The patients with only PE showed a significantly better prognosis than the patients with only PD (P=0.0001) or with PD+PE (P=0.019). The patients who underwent exploratory thoracotomy showed poor survival. There were significant differences in the survival in relation to the extent of the primary tumor. In conclusion, the patients with T1-2 of primary tumor and only a small amount of PE without PD can be expected to show long-term survival after tumor resection.     

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

背景与目的：表皮生长因子受体(epidermal growth factor receptor,EGFR)基因的突变状态是非小细胞肺癌(non-small cell lung cancer,NSCLC)患者使用EGFR酪氨酸激酶抑制剂(tyrosine kinase inhibitors, TKIs)的重要疗效预测指标。该研究旨在探讨突变特异性免疫组织化学(immunohistochemistry,IHC)法检测NSCLC标本EGFR基因突变的临床实用价值。方法：同时采用突变特异性IHC法和扩增阻滞突变系统(amplifi-cation refractory mutation system,ARMS)法检测290例NSCLC患者的EGFR基因突变状态,计算突变特异性IHC法检测EGFR基因突变的灵敏度、特异度、阳性预测值(positive predictive value,PPV)和阴性预测值(negative predictive value,NPV);比较ARMS法和突变特异性IHC法检测EGFR突变的一致性。结果：以ARMS法检测结果为金标准,当染色评分≥1+为阳性时,突变特异性IHC法诊断EGFR基因突变的灵敏度为72.92%,特异度为95.20%,PPV为93.75%,NPV为78.08%。突变特异性IHC法诊断不同类型EGFR基因突变的准确性相差明显：诊断19外显子缺失突变的灵敏度只有55.55%,但其特异度在99%以上;当染色评分为1+时,诊断L858R突变的灵敏度为90.27%,特异度为95.86%,当染色评分为2+或3+时,其特异度则为98.63%~100%。突变特异性IHC法与ARMS法检测结果有较好的一致性(P<0.001,Kappa值：0.612~0.864)。突变特异性IHC法能直观判断EGFR基因突变细胞丰度。结论：突变特异性IHC法是EGFR突变分子检测的有效补充。     

非小细胞肺癌患者血清EGFR基因突变循环DNA检测的临床研究

目的:探讨非小细胞肺癌患者血清EGFR基因突变循环DNA检测的临床意义。方法:选取2011年1月-2014年5月于我院接受治疗的124例非小细胞肺癌患者为研究对象,收集血清及组织标本DNA,检测DNA EGFR基因突变情况。结果:124例非小细胞肺癌患者血清EGFR基因检测结果显示:19号外显子突变11例,21号外显子突变8例,血清总检出19例,检出率为15.3%。组织样本中EGFR基因检测结果显示:19号外显子突变27例,21号外显子突变13例,血清总检出40例,检出率为32.3%。在组织样本中,79例男性EGFR基因突变检出14例,比例为17.7%;45例女性EGFR基因突变检出26例,比例为57.8%,相比具有显著性差异(P<0.05)。76例吸烟史患者检出突变16例,比例为21.1%;48例无吸烟史者检出突变24例,比例为50.0%,两者相比差异显著(P<0.05)。血清样本检测中结果与组织样本基本吻合,在性别和吸烟史方面患者基因突变有显著性差异(P<0.05),在癌症分期及类型方面无显著性差异(P>0.05)。结论:非小细胞肺癌患者血清循环DNA检测EGFR基因突变与肿瘤组织检测具有高度的一致性,这对肺癌患者的诊断、筛查和治疗有着重要的意义,为临床检测提供方便、有效的诊断方法。