肺癌患者呼出气冷凝液检测的研究进展

呼出气冷凝液(EBC)检测是一种非侵入性的诊断技术。检测EBC中微卫星、p53基因、癌胚抗原、内皮素-1等肿瘤标志物,对肺癌筛查、早期诊断、病情监测、疗效评估、跟踪随访等起重要作用。肿瘤标志物的联合检测有助于肺癌的早期诊断。     

肺癌患者呼出气冷凝液检测的研究进展

呼出气冷凝液（EBC）检测是一种非侵入性的诊断技术。检测EBC中微卫星、p53基因、癌胚抗原、内皮素-1等肿瘤标志物，对肺癌筛查、早期诊断、病情监测、疗效评估、跟踪随访等起重要作用。肿瘤标志物的联合检测有助于肺癌的早期诊断。     

呼出气冷凝液中肺癌生物标志物检测的临床意义

 呼出气冷凝液(EBC)检测是近年来新出现的一种检测呼吸道生化成分的新技术,具有无创、简便易行、重复性好等优点。检测肺癌患者EBC中生物标志物,对肺癌筛查、早期诊断、病情监测、疗效及预后评估、随访等起重要作用。     

Exhaled breath condensate pH in patients with lung cancer

Assessment of exhaled breath condensate (EBC) pH is a promising method for investigating and monitoring airway pathology in a number of lung diseases. In this cross-sectional study we tested whether development of lung cancer is associated with acidification of EBC. EBC was collected in 43 smoking patients with lung cancer (squamous cell carcinoma: 17 patients, adenocarcinoma: 16 patients, and small cell lung cancer: 10 patients) before receiving any anticancer treatment and in 20 healthy smokers without any clinical and radiological evidence of pulmonary tumor. EBC pH was measured by CO₂ gas standardization, the most reliable and accurate method at present. EBC pH in patients with pulmonary tumor (6.68 ± 0.02) and in controls (6.63 ± 0.05) was similar (p > 0.05). Results were affected neither by the histological subtype nor the stage of the tumors. Our data suggest that assessment of EBC pH is of limited value for the diagnosis and/or screening of lung cancer.     

Detection of p53 gene mutations in exhaled breath condensate of non-small cell lung cancer patients

Early diagnosis of lung carcinoma is greatly desired. A potential source of early information regarding the process of cancerisation in the airways is exhaled breath condensate (EBC). The direct approach to detecting cancerisation is examining DNA from the area of chronic damage, i.e. airways and lung parenchyma. We therefore investigated DNA in EBC of patients with NSCLC and healthy volunteers. Human DNA was amplified by PCR in exhaled breath condensate and used to detect p53 mutations. A PCR of the beta-actin gene fragment was used to detect human DNA in each of the EBC samples. In 65.7% of the samples, the beta-actin gene was found. Extracted DNA as well as native EBC were equally suited as starting material for amplification. Mutations of the p53 gene were investigated in all EBC samples of NSCLC patients. p53 exons 5-8 were amplified using nested PCR and subsequently sequenced. Mutations were found in four of the patients (n=11; 36.4%) while no mutation was found in volunteers (n=10). Mutations detected in EBC were also compared with those of corresponding tumor tissue. Different point mutations in EBC and tumor tissue were revealed in all cases. Our findings demonstrate that exhaled breath condensate may be used for analysis of somatic gene mutations in an area of direct tobacco-related DNA damage.     

Endothelin-1 is increased in the breath condensate of patients with non-small-cell lung cancer

One recent line of cancer research is currently directed to the study of growth factors. Of increasing interest is endothelin-1 (ET-1), a mitogenic factor already investigated in several human cancer cell lines, which has been found to participate in the development and progression of tumours. This peptide has an important role also in non-small-cell lung cancer (NSCLC) where ET-1 expression has been found in 100% of cell lines. OBJECTIVES: The aim of this study was to measure ET-1 concentrations in the airways of patients with NSCLC using a completely non-invasive procedure--the breath condensate--and to verify the involvement of this peptide in the growth of lung tumours. METHODS: We enrolled 30 patients (17 men, median age 63 years; range 53-74) with histological evidence of NSCLC and 15 healthy controls (9 men, median age 59 years; range 52-70). ET-1 was measured in the exhaled breath condensate by means of a specific enzyme immunoassay kit. RESULTS: Higher concentrations of exhaled ET-1 were found in NSCLC patients (8.3 +/- 0.7 pg/ml) compared to controls (5.2 +/- 0.5 pg/ml, p < 0.0001). A statistically significant difference was observed between patients with distant metastases (stage IV) of NSCLC (8.9 +/- 0.6 pg/ml) and those with locoregional disease (stage I-III) (7.9 +/- 0.5 pg/ml). A significant reduction in ET-1 levels was found in 14 patients after surgical removal of the tumour either associated with or without adjuvant chemotherapy (6.3 +/- 0.5 vs. 7.9 +/- 0.4 pg/ml, p < 0.0001). CONCLUSIONS: These findings suggest that the measurement of ET-1 in the breath condensate of patients with NSCLC could be proposed as a marker for early detection of NSCLC as well as for monitoring reduction or progression of the neoplasm in the follow-up of treated patients.     

Methylation of P16 in exhaled breath condensate for diagnosis of non-small cell lung cancer

Background

Non-small cell lung cancer is the most frequently cause of cancer-related death in the world. To explore the technical feasibility, we detected aberrant promoter methylation of P16 in exhaled breath condensate which was a new, non-invasive tool for diagnosis and screening program of NSCLC.

Methods

We analyzed aberrant promoter methylation of P16 in 180 samples from 60 individuals, including 30 NSCLC patients (cancer tissues, adjacent normal lung tissues, blood plasma, and EBC), and 30 healthy controls (blood plasma and EBC) by fluorescent quantitative methylation-specific polymerase chain reaction (F-MSP).

Results

The positive rate of aberrant promoter methylation of P16 was 26 of 30 (86.66%) in tumor tissues, 15 of 30 (50%) in blood plasma, and 12 of 30 (40%) in EBC, we have not observed the positive methylation of P16 in the adjacent normal lung tissues, or in EBC or blood plasma from the healthy control group.

Conclusion

We found that detected promoter methylation of P16 in EBC was feasibility, it should be an useful biomarker for diagnosis of NSCLC, it have potential prospect that detected the gene molecular in EBC because of noninvasive, specificity, convenient and repeatable.     

非小细胞肺癌患者术后血清VEGF、MMP-9动态变化的研究

背景与目的血管内皮生长因子(vascular endothelial growth factor,VEGF)及基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)在肿瘤的血管生成中起重要作用。研究发现肺癌患者血清VEGF及MMP-9水平升高。本研究旨在监测非小细胞肺癌(non-small cell lung cancer,NSCLC)患者原发肿瘤切除前后的血清VEGF及MMP-9的水平,了解其动态变化规律。方法应用酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测法,分别监测57例NSCLC及18例肺良性病患者术前、术后第1、7、14天血清VEGF及MMP-9的浓度,30例健康志愿者确定血清VEGF及MMP-9正常水平。结果NSCLC患者术前血清VEGF浓度(685.50pg/mL)显著高于肺良性疾病患者术前(160.90pg/mL)及健康人血清水平(94.40pg/mL)(P<0.01);手术切除肿瘤后,NSCLC患者术后第1、7、14天血清VEGF水平持续升高(1055.60pg/mL、1533.90pg/mL、1882.10pg/mL)(P<0.01)。NSCLC患者术前血清MMP-9浓度(84.48ng/mL),显著高于肺良性疾病患者术前(58.98ng/mL)及健康人血清水平(42.94ng/mL)(P=0.000);手术切除肿瘤后,术后第1天NSCLC患者的血清MMP-9水平显著升高(282.99ng/mL),术后第7、14天逐渐降低(221.14ng/mL、194.78ng/mL)(P<0.01)。结论NSCLC患者血清VEGF及MMP-9水平显著高于肺良性病患者及健康人,原发肿瘤切除后血清VEGF、MMP-9水平显著升高。     

肺癌患者血清中血管内皮生长因子的表达及其临床意义

目的:采用ELISA法检测肺癌患者血清血管内皮生长因子（VEGF）含量变化,以探讨其改变与肺癌临床诊断以及预后的关系。方法:原发性肺癌患者50例,取空腹静脉血4ml,采用酶联免疫吸附法(ELISA)测定血清中VEGF的水平,同时检测肺良性病变及健康体检者各50例,分别作为良性对照组及健康对照组。结果:肺癌组VEGF表达水平明显高于肺良性疾病组和健康对照组,有统计学意义（P＜0.05）;小细胞肺癌高于非小细胞肺癌,淋巴结转移者高于无转移者,差异均显著（P＜0.05）;且依TNM分期的早晚呈上升趋势,即:VEGF水平Ⅳ期＞Ⅲ期＞Ⅱ期＞Ⅰ期,组间差异显著（P＜0.05）;而血清VEGF水平与患者生存期呈负相关(r=-0.38,P＜0.05)。结论:检测肿瘤患者血清中VEGF的水平变化,对于肺癌的诊断和预后具有重要意义。     

Influence of thoracic radiotherapy on exhaled nitric oxide levels in patients with lung cancer

BACKGROUND: To determine the physiological role of exhaled nitric oxide (NO) in patients with lung cancer. METHODS: We investigated changes in exhaled NO levels in 29 patients undergoing thoracic radiation therapy with or without chemotherapy. The exhaled NO level was assessed using a chemiluminescence analyzer. RESULTS: The level of exhaled NO was higher in patients with lung cancer before treatment than in controls. With radiotherapy, the exhaled NO level decreased for patients undergoing 40 Gy irradiation and post-radiotherapy. However, five patients showed elevated levels of exhaled NO three times or more than that before radiotherapy. Three of these patients showed signs of radiation pneumonitis. However, none of the other patients showed signs of radiation pneumonitis (p = 0.002). CONCLUSION: Radiation therapy can lower exhaled levels of NO and the levels of exhaled NO may be a useful index for the early prediction of radiation pneumonitis.     

前列腺癌患者血清白细胞介素-18和VEGF相关性

Objective  

The aim of our study was to analyze interleukin-18 (IL-18) and vascular endothelial growth factor (VEGF) serum levels in patients with prostate cancer before and after operation and the possible correlation between IL-18 and VEGF serum levels.     

Detection of the EGFR mutation in exhaled breath condensate from a heavy smoker with squamous cell carcinoma of the lung

A 61-year-old male smoker (40 pack-years) presented with right chest pain. Computed tomography of the chest revealed a cavitary mass in the right lower lobe. A transbronchial biopsy showed squamous cell carcinoma. We examined epidermal growth factor receptor (EGFR) mutations in exhaled breath condensate (EBC). The DNA extracted from his EBC showed a deletion mutation in exon 19. Subsequently, the del E746-A750 mutation in exon 19 in a transbronchial tissue specimen was confirmed. Although he underwent whole-brain irradiation against multiple brain metastases, he had paralysis of the left side of the body and his performance status was 3. The patient was treated with gefitinib. He had marked tumor regression and no symptoms. Although only a small percentage of heavy smokers with squamous cell carcinoma harbor EGFR mutations, they probably benefit from EGFR-tyrosine kinase inhibitors. EGFR mutation status in the patients having such clinical features might be examined.     

Prediction of response to anti-PD-1 therapy in patients with non-small-cell lung cancer by electronic nose analysis of exhaled breath

BackgroundImmune checkpoint inhibitors have improved survival outcome of advanced non-small-cell lung cancer (NSCLC). However, most patients do not benefit. Therefore, biomarkers are needed that accurately predict response. We hypothesized that molecular profiling of exhaled air may capture the inflammatory milieu related to the individual responsiveness to anti-programmed death ligand 1 (PD-1) therapy. This study aimed to determine the accuracy of exhaled breath analysis at baseline for assessing nonresponders versus responders to anti-PD-1 therapy in NSCLC patients.MethodsThis was a prospective observational study in patients receiving checkpoint inhibitor therapy using both a training and validation set of NSCLC patients. At baseline, breath profiles were collected in duplicate by a metal oxide semiconductor electronic nose (eNose) positioned at the rear end of a pneumotachograph. Patients received nivolumab or pembrolizumab of which the efficacy was assessed by Response Evaluation Criteria in Solid Tumors (RECIST) version 1.1 at 3-month follow-up. Data analysis involved advanced signal-processing and statistics based on independent t-tests followed by linear discriminant and receiver operating characteristic (ROC) analysis.ResultsExhaled breath data of 143 NSCLC patients (training: 92, validation: 51) were available at baseline. ENose sensors contributed significantly (P < 0.05) at baseline in differentiating between patients with different responses at 3 months of anti-PD-1 treatment. The eNose sensors were combined into a single biomarker with an ROC-area under the curve (AUC) of 0.89 [confidence interval (CI) 0.82–0.96]. This AUC was confirmed in the validation set: 0.85 (CI 0.75–0.96).ConclusionENose assessment was effective in the noninvasive prediction of individual patient responses to immunotherapy. The predictive accuracy and efficacy of the eNose for discrimination of immunotherapy responder types were replicated in an independent validation set op patients. This finding can potentially avoid application of ineffective treatment in identified probable nonresponders.     

NSCLC患者血清VEGF及VEGFR-2化疗前后的变化及临床意义

目的:探讨NSCLC患者化疗前后血清中VEGF和VEGFR-2的变化、两者相关性及临床意义。方法:用ELISA法检测45例NSCLC患者(化疗组)化疗前后血清中VEGF及VEGFR-2的表达水平,并与20例健康人(对照组)作比较。结果:化疗组化疗前血清VEGF、VEGFR-2表达水平与健康对照组比较明显升高(P<0.01)。3个周期化疗后,部分缓解组VEGF、VEGFR-2表达水平均较化疗前明显降低(P<0.05),降低程度高于稳定组(P<0.05);稳定组化疗后VEGF、VEGFR-2水平较化疗前显著降低(P<0.05);进展组化疗后VEGF、VEGFR-2水平均较部分缓解组和稳定组化疗后显著升高(P<0.05)。化疗前VEGF水平与VEGFR-2表达呈正相关(P<0.05)。结论:NSCLC患者血清中VEGF、VEGFR-2均高表达。测定NSCLC患者化疗前后血清VEGF、VEGFR-2水平的变化有助于临床判断化疗效果及肿瘤进程。     

血清ＴＰＳ、ＶＥＧＦ和ＣＥＡ在非小细胞肺癌化疗疗效评估中的临床价值

目的　探讨非小细胞肺癌（ＮＳＣＬＣ）化疗前后肿瘤标记物水平变化在化疗疗效评价中的临床意义。方法　４１例ＮＳＣＬＣ患者在化疗前后分别测定肺癌特异性的肿瘤标记物,包括组织多肽特异性抗原（ＴＰＳ）、血管内皮生长因子（ＶＥＧＦ）和癌胚抗原（ＣＥＡ）的血清水平,观察其化疗前后的变化,同时在化疗后复查胸部ＣＴ以评价近期疗效,分析两者间的联系。结果　化疗有效（ＣＲ＋ＰＲ）组：血清ＴＰＳ、ＶＥＧＦ和ＣＥＡ水平均有不同程度的下降,但与化疗前相比,仅ＴＰＳ差异具有统计学意义（Ｐ＜０.０５）;化疗无效（ＳＤ＋ＰＤ）组：化疗后血清ＴＰＳ水平明显升高,差异具有统计学意义（Ｐ＜０.０５）,而ＶＥＧＦ和ＣＥＡ水平的变化差异无统计学意义（Ｐ＞０.０５）。结论　化疗前后血清ＴＰＳ水平变化对于判定ＮＳＣＬＣ化疗效果具有较高的临床价值。     

非小细胞肺癌患者手术前后血清中内皮抑素和VEGF水平变化及相关性研究

目的:探讨非小细胞肺癌(non-smallcell lung cancer,NSCLC)患者手术前后血清中内皮抑素(endostatin)和血管内皮生长因子(vasular endothelial growth factor,VEGF)水平的动态变化规律及两者水平变化的相关性。方法:用ELISA法检测46例NSCLC患者手术前后血清中endostatin和VEGF的水平。结果:1)NSCLC患者术后7d血清endostatin水平为(23·41±5·12)ng/mL,显著高于术前[(20·85±4·56)ng/mL]和术后1d[(18·89±4·67)ng/mL]血清endostatin水平,P值分别为0·011和0·000;术后7d血清VEGF水平为(3·75±0·71)ng/mL,显著高于术前[(1·72±0·46)ng/mL]和术后1d[(2·22±0·58)ng/mL]血清VEGF水平,P值均为0·000。2)NSCLC患者术前血清endostatin与VEGF水平呈非常显著负相关,r=-0·380,P=0·009。3)NSCLC患者术后7d血清endostatin水平与VEGF水平呈非常显著正相关,r=0·351,P=0·017。结论:NSCLC患者手术前、后血清endostatin和VEGF水平存在动态变化,且两者手术前后的水平变化有显著相关性,检测NSCLC患者手术前后血清中endostatin和VEGF水平可能是进一步预测肺癌恶性行为的有用指标。肿瘤防治杂志,2005,12(19):1441-1444     

DTC患者血清及组织中CYFRA21.1、Gal-3、VEGF的表达水平及其意义

目的探讨分化型甲状腺癌(differentiated thyroid cancer,DTC)患者血清与组织中细胞角蛋白19片段(cytokeratin fragment antigen 21.1,CYFRA21.1)、半乳凝素-3(galectin-3,Gal-3)、血管内皮生长因子(vascular endothelial growth factor,VEGF)的表达水平及临床意义。方法收集甲状腺肿瘤患者133例,分为:良性疾病组43例:甲状腺腺瘤11例、结节性甲状腺肿32例;恶性疾病组90例:甲状腺滤泡状癌(follicular thyroid carcinoma,FTC)9例,甲状腺乳头状癌(papillary thyroid carcinoma,PTC)71例,PTC伴颈部淋巴结转移(lymph node metastasis,LNM)10例。另设健康对照组30例。采用电化学发光法(electrochemiluminescence immunoassay,ECLIA)、酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)检测血清CYFRA21.1、Gal-3、VEGF水平,免疫组织化学SP法检测甲状腺手术标本中细胞角蛋白19(cytokeratin 19,CK19)、Gal-3、VEGF蛋白的表达。结果血清CYFRA21.1、VEGF水平在各亚组间差异均无统计学意义(均P>0.05)。甲状腺肿瘤患者血清Gal-3水平较对照组有所增高(P<0.01),但良、恶性疾病组间比较差异无统计学意义(P=0.17)。CK19、Gal-3、VEGF在恶性疾病组中表达阳性率分别为88.9%(80/90)、91.1%(82/90)、85.6%(77/90),同良性疾病组比较差异均有统计学意义(均P<0.01)。CYFRA21.1、Gal-3、VEGF血清水平与组织表达皆无相关性(r=-0.094,r=-0.016,r=0.020,均P>0.05)。结论 CK19、Gal-3、VEGF为分化型甲状腺癌可靠的组织标志物,但其血清水平不能用于甲状腺肿瘤的良、恶性鉴别。