Benign Pulmonary Diseases Research Articles

Significance of RUNX3 Hypermethylation in Serum DNA of Non-small Cell Lung Cancer Patients

Objective To detect the promoter hypermethylation status of runt-related transcription factor 3(RUNX3) gene in serum DNA of non-small cell lung cancer patients and to discuss its clinical significance.Methods Serum DNA was extracted from 62 patients with NSCLC 30 patients with benign pulmonary disease,and 16 health subjects with medical check-up.The methylation status of RUNX3 gene was detected by MSP and the correlation of methylation profiles with clinical characteristic was statistically analyzed.Results Aberrant promoter methylation of RUNX3 was found in 25 of 62(40.32%) NSCLC patients,but in none of patients with benign pulmonary disease and health subjects with medical check-up(Fisher exact probabilities test,P0.05).The promoter hypermethylation of RUNX3 correlated with histopathology(χ2=4.12,P0.05)and clinical stages(Fisher exact probabilities test,P0.05),but not with age,gender,smoking index or differentiation in NSCLC patients.Conclusion The promoter hypermethylation of RUNX3 may play an important role in the tumorigenesis and progression.It is hopeful to become a promising novel biomarker for diagnosis in NSCLC.

The diagnostic value of circulating stanniocalcin‐1 mRNA in non‐small cell lung cancer

Previous studies have suggested that the aberrant expression of Stanniocalcin-1 (STC1) occurs in tumor cells. In this study, we explored whether the circulating STC1 mRNA is a promising biomarker in the peripheral blood in patients with non-small cell lung cancer (NSCLC). The level of circulating STC1 mRNA was determined with a sensitive quantitative real-time reverse transcription PCR assay. and the data were analyzed by the statistical methods of one-way ANOVA, Mann-Whitney-Wilcoxon U-Test, and Receiver operating characteristic (ROC) curve analysis. The level of circulating STC1 mRNA in patients with NSCLC was significantly higher than in patients with benign pulmonary disease (P < 0.001) or healthy volunteers (P < 0.001). Higher levels of circulating STC1 mRNA were associated with more advanced tumor stages and histological subtypes. Using a cutoff of 0.055, the sensitivity and specificity of STC1 mRNA levels to differentiate between patients with NSCLC and patients with benign pulmonary diseases was 66.7 and 90.9%, and it was 63.7 and 99.8% for patients with NSCLC and healthy volunteers, respectively. These findings support our hypothesis that circulating STC1 mRNA is a promising biomarker in the peripheral blood.

Elevated circulating levels of osteopontin are associated with metastasis in advanced non-small cell lung cancer

To investigate the relationship between postoperative metastasis and circulating levels of osteopontin in non-small cell lung cancer (NSCLC). The expression of osteopontin mRNA were detected with RT-PCR technique. The circulating levels of osteopontin were measured through ELASA in 46 NSCLC cases that had not been received any anti-cancer treatment at the time of sampling. The tissues from fifteen patients with benign pulmonary diseases were studied as control group. The overall median mRNA expression level of osteopontin was approximately 70-fold higher in tumor tissues than in matched normal lung tissues (P<0.001). Over-expression of osteopontin mRNA was significantly associated with clinical stage (P=0.009). Advanced disease states had higher circulating level of osteopontin (stage I+II versus stage III+VI). In multivariate analysis, stage was the only independent factor influencing circulating levels of osteopontin. All patients were followed up for 12 months, 2 of the 46 patients with both osteopontin mRNA expression and elevated plasma osteopontin levels had local recurrence and 10 had distant metastasis. There was a significant difference in the osteopontin levels between metastasis group and non-metastasis group. Preoperative plasma levels of osteopontin are significantly associated with post-operative metastasis in advanced NSCLC.

Application of serum levels of pro-gastrin releasing peptide, tissue polypeptide specific antigen and neuron specific enolase in therapy monitoring in small cell lung cancer patients

Objective To evaluate the clinical significance of serum levels of ProGRP, TPS and NSE in diagnosis and therapy monitoring in small cell lung cancer patients. Methods The levels of serum ProGRP, TPS and NSE in 51 SCLC patients (SCLC group), 60 benign pulmonary disease patients (benign disease group ) and 60 healthy people (healthy group ) were determined using chemiluminescent immunoassay, ELISA and electrochemiluminescent immunoassay respectively. Blood samples were collected and detected prior to therapy, before the second course of chemotherapy and the third course of chemotherapy consecutively in all the 51 SCLC patients. Results The serum ProGRP, TPS and NSE concentrations prior to chemotherapy in limited stage SCLC (LSCLC) were 136. 9(22.8-631.7)ng/L, 78. 2(56.4-114.6) U/L and 28.1(20.9-46.1)μg/L, respectively; And in extensive stage SCLC patients (ESCLC) were 1 106.6(41.2-2161.1) ng/L, 230. 9( 143.5-259.0) U/L and 81.1 (34.3-140.0)μg/L, respectively. The serum concentrations of the 3 markers in benign disease group were 19. 7 ( 9. 5-29. 1 )ng/L, 48. 7 ( 17.9-95.4) U/L and 12. 1(1.2-13.9) μg/L; and in healthy group were 20.3(10.7-30.6) ng/L, 50.3(19.5-70.7) U/L and 11.7 (1.1-13.4)μg/L, respectively. The Kruskal-Wallis test showed significantly statistical difference in different groups of the 3 tumor markers, Chi-Square were 51. 368,36. 532 and 81. 645( P ＜0. 01 ). Significant statistically differences showed when the concentrations of the 3 marks of the 2 control group were compared with that of the LSCLC group ( U =491, 827, 609 and 476, 831, 585,respectively, P ＜ 0. 05 ). Differences were also statistically significant when the 2 control group compared with that of the ESCLC group ( U = 314,532,456 and 302,553,430, respectively, P ＜ 0. 01 ). The AUC of ProGRP was 0.832 +0.029(95% CI:0.774-0.890). When cutoff value of ProGRP set as 37.7 ng/L, the diagnostic sensitivity, specificity, positive predictive value, negative predictive value and Youden's index were 71% (36/51), 97% (116/120), 90% (36/40), 89% ( 116/131 ) and 67%, respectively; show good detection performance. The sensitivity increased to 92%, 86%, 92% and 88%, when combination detection of ProGRP + TPS + NSE, ProGRP + TPS, ProGRP + NSE and TPS + NSE were used, and the specificities were 77%, 77% , 92% and 77% accordingly. The Fridman test showed significantly statistical difference in the 3 tumor markers at different stages of treatment, x2 were 49. 120, 10. 614 and 44. 392, P ＜0. 01. After the first chemotherapy course, all the tumor marker levels except TPS decreased significantly in comparison with the pretreatment concentrations. However, only ProGRP levels showed a progressive drop during the two consecutive courses of therapy, and the median concentrations were 68.0 ( 18. 6-158.4 ) and 21.0( 14. 9-63.5) ng/L (compared to the level before therapy,Z=-4. 889 and -5. 594, P ＜0. 01 ). The median of serum TPS increased slightly to 105.2 (54. 1-181.2 ) U/L after the first chemotherapy course (Z=-1.248, P＞0.05), and decreased significantly to 79.0(48.7-155.3) U/L after the second chemotherapy course (Z=-2.484, P＜0. 05 ). As to the NSE, the median concentration decreased to 11.8(8.0-16.0)μg/L after the first chemotherapy course ( Z= - 5. 568, P ＜ 0. 01 ). However, the median was 10. 6(9.0-12.7)μg/L, which showed no significant decrease after the second chemotherapy course (Z=-1.851, P＞0.05).Forty-six SCLC patients evaluated as clinical remission ( 3 CR and 43 PR) after the second chemotherapy course, among them there were 38 patients (83%) with normal serum ProGRP, TPS and NSE level ( 19 patients) or with only 1 abnormal tumor level ( 19 patients). There were only 2 patients with all abnormal serum ProGRP, TPS and NSE level, and both patients were evaluated as clinical PD. Two patients with 2 abnormal tumors results were classified as SD, the only 1 patient without therapy evaluation also had 2 abnormal tumor marker results. Conclusions The serum ProGRP, TPS and NSE are valuable tumor markers for diagnosis and treat monitoring of SCLC, particularly the ProGRP + NSE shows the highest clinical value. Combing detection of the 3 tumor markers are valuable for therapy monitoring and prognosis in SCLC patients. Key words: Lung neoplasms; Carcinoma, small cell; Peptide fragments; Phoshopyruvate hydratase

Plasma proGRP Concentration is Sensitive and Specific for Discriminating Small Cell Lung Cancer from Nonmalignant Conditions or Non-small Cell Lung Cancer

To date, most clinical data on pro-gastrin-releasing peptide (proGRP) have been based on serum concentrations. This study evaluated the agreement between proGRP levels in fresh serum and plasma in patients with various lung diseases. Pairs of serum and EDTA plasma were collected from 49 healthy individuals. At the same time, EDTA plasma of 118 lung cancer patients and 23 patients with benign pulmonary diseases were prospectively collected. Compared to serum, plasma proGRP concentrations were higher by an average of 103.3%. Plasma proGRP was higher in malignancy (336.4 ± 925.4 pg/mL) than in benign conditions (40.1 ± 11.5 pg/mL). Small cell lung cancer (SCLC) patients showed higher levels of proGRP (1,256.3 ± 1,605.6 pg/mL) compared to other types of lung cancer. Based on the ROC curve analyses at a specificity of 95%, the diagnostic sensitivity of plasma proGRP was estimated to be 83.8% in distinguishing SCLC from all the other conditions, and 86.5% for discriminating SCLC from the nonmalignant cases. Among the SCLC cases, limited stage disease had lower levels of plasma proGRP than extensive disease. When measuring circulating levels of proGRP, the use of plasma is preferred over serum. Plasma proGRP has a potential marker for discriminating SCLC from nonmalignant conditions or non-small cell lung cancer.

The study of differential proteins in patients with peripheral lung cancer by surface-enhanced laser desorption/ionization time of flight mass technology

To scan the protein mass spectra in the sera and bronchoalveolar lavage fluid (BALF) from patients with peripheral lung cancer, screen out the differential proteins, and explore the clinical significance of the differential proteins. SELDI-TOF-MS was used to detect the protein mass spectra and to screen out the differential proteins in the sera and BALF collected before and after lung biopsy in 20 patients with peripheral lung cancer and 20 patients with benign pulmonary diseases. The differential proteins were analyzed and the initial diagnostic models were set up. (1) There were 6 differential protein peaks in the sera of the 2 groups (P < 0.05). The protein with a mass/charge ratio (M/Z) of 6637 was selected to establish the diagnostic model. The sensitivity of diagnosing peripheral lung cancer was 70% (14/20), the specificity 90% (18/20), the accuracy 80% (32/40), the positive predictive value (PV+) 88% (14/16), the negative predictive value (PV-) 75% (18/24), and the area under the ROC curve (AUC) was 0.73. (2) There were 11 differential protein peaks in the BALF collected before lung cancer biopsy of the 2 groups (P < 0.05). The protein with a M/Z of 7982 was selected to establish the diagnostic model. The sensitivity of diagnosing peripheral lung cancer was 85% (17/20), the specificity 90% (18/20), the accuracy 88% (35/40), the PV+ 89% (17/19), the PV- 86% (18/21), and the AUC was 0.94. (3) There were 14 differential protein peaks in the BALF collected after lung cancer biopsy of the 2 groups (P < 0.05). The protein with a M/Z of 7671 was selected to establish the diagnostic model. The sensitivity of diagnosing peripheral lung cancer was 85% (17/20), the specificity 100% (20/20), the accuracy 93% (37/40), the PV+ 100% (17/17), the PV- 87% (20/23), and the AUC was 0.93. There were more differential proteins in BALF as compared with sera. There were more differential proteins in the BALF collected after lung biopsy as compared to that before lung biopsy. The AUC of the diagnostic models set up by proteins in BALF collected before and after lung biopsy were all above 0.9 and showed higher efficiency for the diagnosis of peripheral lung cancer as compared to proteins in sera. These differential proteins may be better tumor markers for the diagnosis of peripheral lung cancer at the early stage.

The correlation between the expression of EGFR in tumor tissue and in peripheral blood in non-small cell lung cancer patients

背景与目的表皮生长因子受体（epidermal growth factor receptor, EGFR）在肿瘤细胞的增殖中起重要作用，肿瘤组织中EGFR表达水平可为肺癌的诊断、预后判断提供依据。本研究通过检测EGFR在非小细胞肺癌患者肿瘤组织和外周血中的表达，探讨利用外周血中EGFR表达水平来预测肿瘤组织中的表达并作为肿瘤标记物的可行性。方法应用real-time RT-PCR技术检测46例非小细胞肺癌组织和10例肺良性病变组织中EGFR mRNA的表达；应用ELISA法检测46例非小细胞肺癌患者外周血中EGFR蛋白的表达，并以10例肺良性疾病患者的外周血作对照。结果直线相关分析表明EGFR在外周血中的蛋白表达水平与肿瘤组织中的mRNA表达水平具有相关性（R2=0.83, P=0.016）。结论血清中EGFR蛋白水平的测定有可能替代肿瘤组织中mRNA的检测，为无法取得肿瘤组织或取到肿瘤组织困难的患者提供一种更快捷、简便的替代检测方法，尽早为临床选择治疗方法及预后判断提供依据。

Diagnosis value of the detection of CYFRA21-1 in non-small cell lung cancer

背景与目的细胞角蛋白19片段（cytokeratin 19 fragment, CYFRA21-1）是血液中的一种可溶性蛋白质，它可能是一种潜在的循环肿瘤标志物。本研究旨在探讨CYFRA21-1在非小细胞肺癌（non-small cell lung cancer, NSCLC）诊断中的应用价值。方法采用酶联免疫吸附法分别检测107例NSCLC患者和51例肺部良性疾病患者外周血中CYFRA21-1含量，应用受试者工作曲线（receiver operaing characteristic curves, ROC）对检测结果进行临床评价。结果NSCLC组织与肺良性疾病组外周血CYFRA21-1水平差别有统计学意义（χ2=47.343, P < 0.001）；以3.3 μg/L为诊断临界点，CYFRA21-1诊断NSCLC的敏感性和特异性分别为74.77%和76.47%，ROC曲线下面积为0.813 9；NSCLC各病理类型之间CYFRA21-1水平差别无有统计学意义（χ2=0.450, P=0.799）；外周血CYFRA21-1在广泛期（Ⅲb、Ⅳ）病例中明显高于局限期（Ⅰ、Ⅱ、Ⅲb）病例（χ2=7.057, P=0.008）。结论CYFRA21-1作为肺癌肿瘤标志物具有相对较高的敏感性和特异性；血浆中CYFRA21-1水平升高提示肺癌患者可能已经发展为广泛期。

Simultaneous operation of concomitant diseases in the esophagus and lung

The aim of this study was to retrospectively review our experience of performing simultaneous operations on concomitant diseases in the esophagus and lungs. From January 1998 to July 2009, simultaneous operations were performed on 13 patients with concomitant esophageal and pulmonary diseases, using coordinated surgical approaches. Among the 13 patients, six had primary cancers in the esophagus and lungs, five had primary esophageal cancer accompanied by a benign pulmonary disease, one had benign diseases in both esophagus and lung, and one had primary esophageal cancer with metastasis to the left lower lung. All patients survived the operations. Two major complications occurred postoperatively. One complication was bronchopleural fistula and the other was intrathoracic gastric laceration. Both patients recovered after additional treatments. Simultaneous operation of concomitant diseases in the esophagus and lungs is feasible and safe in selected patients who have received careful preoperative assessment, well-designed surgical approach, and proper perioperative management.

Overexpression of IL-8 and MMP-9 confer high malignant phenotype in patients with non-small cell lung cancer

背景与目的白细胞介素-8（interleukin-8, IL-8）曾被认为是一种具有趋化作用的炎症因子，近年的研究认为它和基质金属蛋白酶-9（matrix metalloproteinase-9, MMP-9）均为与肿瘤生长和转移密切相关的细胞因子，本研究通过检测IL-8、MMP-9在非小细胞肺癌患者组织和血清中的表达，研究二者与临床病理特征之间的联系，分析肺癌组织和血清中IL-8、MMP-9表达的相关性，并探讨IL-8、MMP-9作为肿瘤标志物评估非小细胞肺癌患者病情进展程度的效能。方法应用酶联免疫吸附实验（enzyme-linked immunosorbent assay, ELISA）检测141例非小细胞肺癌患者、40例健康人和40例肺良性疾病患者血清中IL-8、MMP-9的水平。采用免疫组化SP法检测95例非小细胞肺癌、21例良性疾病肺组织及25例正常肺组织中IL-8和MMP-9的表达。结果IL-8、MMP-9在非小细胞肺癌患者血清和组织的表达水平明显高于肺良性疾病对照组和健康对照组，其差异有统计学意义，且随着临床病理分期的升高而升高。在肺癌患者组织和血清中，IL-8与淋巴转移均有较强的相关性，在肺癌组织中IL-8和MMP-9的表达具有很强的相关性（r=0.765）。结论IL-8、MMP-9表达水平与非小细胞肺癌的进展密切相关，特别是IL-8与肺癌的淋巴转移具有明显的相关性，其机制可能是通过上调MMP-9实现的。

The correlation between the expression of Wnt1 and prognosis in resected non-small cell lung cancer

背景与目的Wnt1蛋白是Wnt信号传导通路的第一个因子，其高表达与很多肿瘤相关，本研究旨在探讨Wnt1蛋白在非小细胞肺癌组织（non-small cell lung cancer, NSCLC）中的表达及与预后的相关性。方法选取术后经病理证实的115例NSCLC和19例肺良性病变（5例肺结核、4例支气管扩张、6例肺大疱、4例炎性假瘤），运用免疫组化Envision法检测Wnt1蛋白的表达，采用χ2检验分析Wnt1蛋白在NSCLC及良性组织中表达的差异及NSCLC中Wnt1蛋白表达与临床病理特征的相关性。采用Kaplan-Meier生存分析和Cox回归分析方法分析Wnt1蛋白在NSCLC组织中的表达和预后的关系。结果Wnt1在NSCLC组织中表达的阳性率为62.6%，显著高于对照组的31.6%（χ2=4.474, P=0.034），但与临床病理特征无相关性。Kaplan-Meier生存分析、Log-rank检验提示Wnt1阳性表达的NSCLC的患者预后较差（P=0.003），Cox回归分析结果表明，Wnt1蛋白是影响NSCLC预后的独立危险因素（OR=1.834, P=0.032）。结论Wnt1蛋白在NSCLC组织中的阳性表达率高于肺良性病变组织；Wnt1蛋白阳性表达者预后差，可以作为判断NSCLC预后的参考指标。

210 Identification and characterization of amphiregulin as a new biomarker of resistance to gefitinib in non-small cell lung cancers

RASSF1A is a novel putative tumor suppressor gene located in 3p21.3 region. The most common inactivation mechanism of RASSF1A is promoter hypermethylation, which is observed in multiple solid tumors including lung cancer. In the present study, we identified the methylation status of RASSF1A in lung cancer sera using methylation-specific PCR and analyzed its clinicopathological significance. Hypermethylation of RASSF1A was detected in 27 of 80 (33.8%) cancer patients but no benign pulmonary disease patients or healthy donors (P < 0.001). RASSF1A hypermethylation was preferentially observed in small cell lung cancer (P = 0.042), while no statistical difference was found among methylation frequencies of different subtypes of non-small cell lung cancer. RASSF1A methylation status was associated with differentiation (P = 0.009) and stage (P = 0.013), but not with gender, age or treatment. These findings suggest that serum RASSF1A hypermethylation is a promising molecular biomarker for lung cancer diagnosis.

Expression of CDK5/p35 in resected patients with non-small cell lung cancer: relation to prognosis

Overall outcome of those patients with non-small cell lung cancer (NSCLC) remains poor. Recently, several studies demonstrated that cyclin-dependent kinase-5 (CDK5) activity with its specific activator protein p35 was important for spontaneous metastasis in various types of carcinomas. Our objective was to explore the expression of CDK5 and its prognostic indicator in patients with NSCLC. Immunofluorescent staining was used to detect the expression of CDK5/p35 in the lung tissue of 95 patients with NSCLC and 20 patients with benign pulmonary disease. The correlation between the expression of CDK5/p35 and clinicopathologic features of patients with NSCLC was investigated. The 5-year overall survival of patients with tumors expressing different levels of CDK5/p35 was evaluated by the Kaplan-Meier method. Positive expressions of CDK5/p35 were detected in the tumor cells in 66 samples (69.5%) of the 95 patients with NSCLC. Although no remarkable correlation between CDK5/p35 expression and age at the time of surgery, gender, and histopathologic type, there were significant differences between CDK5/p35 expression and degree of differentiation, pathological stage and lymph node metastasis in patients with NSCLC. In addition, we demonstrated that median survival for patients with and without CDK5/p35 expression was 24 and 58 months, respectively, and 5-year overall survival rate 25.8 and 48.3%, respectively (P<0.05). Patients with lung cancer with a positive CDK5/p35 expression had a poorer prognosis than those with a negative CDK5/p35 expression. Based on our results, CDK5/p35 may represent a biomarker for prognosis in patients with NSCLC.

CA 19-9 in patients with benign pulmonary diseases

Although serum CA 19-9 is considered to be a useful and specific tumor marker for pancreatic cancer, some patients with benign pulmonary diseases show elevated serum CA 19-9 levels. We measured serum CA 19-9 levels of 156 patients with benign pulmonary diseases (55 with asbestosis, 11 with bronchial asthma, 32 with bronchiectasis, 16 with idiopathic pulmonary fibrosis (IPF), 13 with healed pulmonary tuberculosis (HPT) and 29 other benign diseases). The percentage of patients with positive serum CA 19-9 was 42.3% (14.5% in asbestosis, 27.3% in bronchial asthma, 59.4% in bronchiectasis, 81.3% in IPF, 61.5% in HPT and 51.7% in others). In some patients, serum CA 19-9 levels were as high as those found in malignant gastrointestinal diseases. Serum CA 19-9 levels correlated well with disease activity. Immunohistochemically, CA 19-9 was expressed in mucous cells of the bronchial gland and surface of the bronchiolar surface epithelium cells in benign pulmonary disease. Gel filtration study suggested some difference in molecular weight between the serum CA 19-9 antigen of lung cancer and that of benign pulmonary diseases. It is suggested that serum CA 19-9 increases in the case of hyperplasia of the bronchiolar epithelium cells or the mucous cells of the bronchial gland. We conclude that benign pulmonary disease is one of the factors that affect serum CA 19-9 levels.

Plasma DNA level as a tumor marker in primary lung cancer

The plasma level of human DNA was determined by the dot-hybridization method using human Alu-family DNA as a probe in 45 patients with primary lung cancer, 54 patients with benign pulmonary diseases, and 59 healthy controls. The mean plasma DNA level was significantly higher in the patients with lung cancer than that in the patients with benign pulmonary diseases or in healthy controls. The mean plasma DNA level in the patients with benign pulmonary diseases was also significantly higher than that in healthy controls. There was no significant difference in mean plasma DNA level in each histologic type of lung cancer. The plasma DNA level was elevated above the cut-off level of 80 ng/ml in 71% of the patients with lung cancer, 37% of the patients with benign pulmonary diseases and none of the healthy controls. The serum CEA was positive in 38% of the patients with lung cancer and thus when plasma DNA and serum CEA were used in combination, 78% of the cases with lung cancer could be detected by these two markers. In the patients with lung cancer who responded to treatment, the plasma DNA levels were significantly decreased after treatment, while its levels were elevated in the patients whose treatment was unsuccessful. These findings indicate that plasma DNA may be a useful marker in patients with lung cancer.

Serum lipid-bound sialic acid as a marker in lung cancer patients

To evaluate the usefulness of serum lipid-bound sialic acid (LSA) as a tumor marker in patients with lung cancer, its serum levels were determined in 85 patients with primary lung cancer, 42 patients with benign pulmonary diseases and 22 healthy controls by the method developed by Katopodis and Stock. The mean serum LSA concentration was significantly higher in the patients with primary lung cancer [32.4 +/- 16.4 (SD) mg/dl] than that in the patients with benign pulmonary diseases (22.4 +/- 12.4 mg/dl) or in healthy controls (17.2 +/- 4.5 mg/dl). The serum LSA level was elevated above the mean value plus 2 SD for the healthy controls (26.0 mg/dl) in 60% of all cases with primary lung cancer including 1 of 9 (11.1%) in stages I and II and 50 of 76 (65.8%) in stages III and IV and 35.4% of the patients with benign pulmonary diseases. No significant difference was observed in serum LSA concentration among each cell type of lung cancer. Serum carcinoembryonic antigen (CEA) was elevated (greater than 5.0 ng/ml) in 39.8% of the patients with lung cancer. As there was poor correlation between serum LSA and CEA levels, when these two markers were used in combination, 71.1% of the cases with lung cancer in this study showed elevated levels of either serum LSA or CEA, or both. Moreover, in the patients with small cell lung cancer followed up during intensive chemotherapy, serum LSA levels changed almost parallel with the clinical course. These results indicate that serum LSA may be a useful biochemical marker in lung cancer patients.

Diagnotic value of the combined determination of telomerase activity in induced sputum, pleural effusion and fiberobronchoscopic biopsy samples in lung cancer

背景与目的已有的研究表明, 端粒酶的异常活化与肺癌的发生、发展及转归密切相关, 端粒酶是目前已知广谱的肿瘤分子标记物, 是诊断和治疗肺癌的重要分子标志物之一。本研究旨在探讨诱导痰、胸水和纤维支气管镜活检组织中端粒酶活性的联合检测对肺癌的诊断价值。方法对80例肺癌伴胸水患者和50例肺部良性病变伴胸水患者, 应用TRAP-PCR-ELISA方法分别检测其诱导痰、胸水和纤维支气管镜活检组织中端粒酶的活性。结果肺癌伴胸水患者诱导痰、胸水和纤维支气管镜活组织中端粒酶的活性均明显高于肺部良性病变伴胸水患者(P < 0.001), 不同病理类型的肺癌患者间三种标本的端粒酶活性差异无统计学意义(P > 0.05)。诱导痰、胸水和纤维支气管镜活组织中端粒酶活性的检测对肺癌的诊断敏感性分别为62.5%(50/80)、46.3%(37/80)和60.0%(48/80);特异性分别为72.0%(36/50)、66.0%(33/50)和70.0%(35/50);准确性分别为66.2%(86/130)、53.8%(70/130)和63.8%(83/130);三项联合检测的敏感性、特异性和准确性分别为85.0%(68/80)、78.0%(39/50)和82.3%(107/130), 其中敏感性与单独诱导痰、胸水和纤维支气管镜活组织的敏感性差异有统计学意义(P < 0.01)。结论诱导痰、胸水和纤维支气管镜活组织端粒酶活性的联合检测具有更高的敏感性, 可以提高肺癌的诊断率。

CK19 mRNA expression in the bone marrow of patients with esophageal squamous cell carcinoma and its clinical significance

The 5-year survival rate in resectable patients with esophageal cancer is only 20% to 36%. Regional relapse and distant metastasis are responsible for the failure of treatment and the majority of cancer-related deaths. Earlier detection of metastases, especially micrometastases, has the potential for more accurate risk stratification in subsequent therapy decisions. No effective techniques have yet been found to detect metastases in erroneously thought to have early stage disease. This study was designed to investigate the clinical significance of bone marrow micrometastases detected by reverse transcriptase-polymerase chain reaction (RT-PCR) in patients with esophageal cancer. Expression of CK19 mRNA in the bone marrow of 61 patients with esophageal squamous cell carcinoma (ESCC) and 15 benign pulmonary and esophageal disease patients was assessed via RT-PCR. Correlation of CK19 mRNA expression to the clinicopathologic features and prognosis of the 61 patients was analyzed: 21.3% (13/61) were positive for expression of CK19 mRNA in patients with ESCC. No CK19 mRNA was detected of the 15 benign pulmonary and esophageal disease patients. CK19 mRNA expression did not correlate with the clinicopathologic features of the patients with ESCC, but patients with CK19 mRNA-positive bone marrow had earlier recurrence and shorter survival after surgery. In multivariate analysis, CK19 mRNA was found to be an independent predictor of a poor outcome. CK19 mRNA may be used as a molecular maker to detect bone marrow micrometastases in patients with ESCC and may help to select the proper therapy and predict the prognosis.

Transcriptional inactivation of secreted frizzled-related protein 1 by promoter hypermethylation as a potential biomarker for non-small cell lung cancer

Epigenetic silencing of secreted frizzled-related protein (SFRP) genes, antagonists of the WNT pathway, contributes to the pathogenesis of several cancers including non-small cell lung cancer (NSCLC). We hypothesize that methylation analysis of SFRPs family could improve their use as a panel of biomarkers for diagnosing and staging of NSCLC in China. The expression of four SFRP members (SFRP1, 2, 4, and 5) in NSCLC samples was screened by RT-PCR and quantitative real-time PCR. Only SFRP1 was significantly downregulated in NSCLC, as compared to adjacent normal tissues and benign pulmonary disease tissues (P=0.006). Promoter hypermethylation of SFRP1 was found in 32.1% (25/78) NSCLC specimens and was closely correlated with loss of expression, besides SFRP1 hypermethylation was associated with lymph metastasis (P=0.039) and disease progression within one year (P=0.027). Furthermore, methylated SFRP1 was detected in 28.2% (22/78) of plasma samples from NSCLC patients while only 4% (2/50) in cancer-free controls, and the concordance of SFRP1 methylation status in tumor tissues and corresponding plasmas was satisfactory (P <0.001). In conclusion, epigenetic inactivation of SFRP1 is a common event contributing to lung carcinogenesis and maybe used as a potential biomarker for NSCLC in Chinese population.

Expression and clinical significance of TTF-1 and p63 in NSCLC.

To detect the expressions of thyroid transcription factor-1 (TTF-1) and p63 in non-small cell lung cancer (NSCLC) and to evaluate their clinical significance. The expression of TTF-1 and p63 from 404 NSCLC and 28 benign pulmonary disease (BPD) tissue specimens were detected by immunohistochemical EnVision two-step method, together with their clinicopathologic data. The positive rate of TTF-1 and p63 protein in NSCLC tissues was 51.7% (209/404) and 37.9% (153/404), respectively, while negative in the BPD group. There was overexpression of TTF-1 in female gender and non-smoking history (P<0.001) and asymptomatic patients (P=0.015). It was more frequently in adenocarcinoma (AdC) with sensitivity of 84.1% and specificity of 89.8%, especially in well or moderately differentiated AdC (P<0.001). The positive rate of p63 was closely related with male gender and smoking history (P<0.001). Its sensitivity and specificity to squamous cell carcinoma (SCC) was 95.5% and 98.8%, respectively, which was positively correlated with differentiation of SCC (P=0.008), but negatively with tumor stage (P=0.002). Logistic multivariate analysis showed smoking history and histological type were significantly associated with TTF-1 and p63 expression. 93.1% of those represent TTF-1(+)/p63(-) were AdC, while 98.6% of TTF-1(-)/p63(+) were SCC. p63 expression was negatively correlated with TTF-1 (P<0.001). TTF-1 and p63 might be effective bio-markers for AdC and SCC in NSCLC. They may be a useful marker panel for the identification and differential of lung cancer.