Early Diagnosis Of Non-small Cell Lung Cancer Research Articles

Detection of Anaplastic Lymphoma Kinase (ALK) gene in Non-Small Cell lung Cancer (NSCLC) By CISH Technique

Background/Aim: Anaplastic Lymphoma Kinase (ALK) rearrangement has evaluated activity of NSCLC compared with other molecular subtypes (EGFR, KRAS). Many studies demonstrated that patients with ALK rearrangement positive NSCLC have improved with a good response rates and progression (free survival) when treated with either monotherapy or by a combination therapy compared with EGFR-mutated, KRAS/EGFR/ALK wild type or KRAS-mutated. The aim of this study was to detect and study the signal pattern of normal ALK and compare it to that of mutated ALK with gene rearrangement in cases of non-small cell lung cancer and Inflammatory conditions by implementing the CISH technique. In addition to correlate ALK signal pattern with the histopathological type and grade as well as the age and sex of the patients.Materials & Methods: Forty patients with NSCLC and Inflammatory diseases were enrolled in a comparative cross sectional study. The tissue blocks were sectioned on non-charged slides for the preparation of routine H&E staining. Positively charged slides were used for tissue sections prepared for chromogenic in situ hybridization procedure to detect ALK gene.Results: ALK gene signal break apart was detected in (18/20, 90%) of malignant cases; (0/20, 0%) of non-neoplastic lung lesions. There was a significant statistical difference in their distribution, p < 0.05. While There was no significant association between any disease status and sex P value = 1.000NS. The CISH test was 100% sensitive using negative score as a cutoff point and 90.9% specific. The score was divided into three levels that categorized the cases, so there were six cases in score one (1-32%), nine cases in score two (33-67%) and three cases in score three (68-100%).Conclusion: Detection of ALK rearrangement in the early diagnosis of NSCLC is highly sensitive and can save a lot of efforts in planning chemotherapy regimens. Results were very promising in identifying this mutation by a sensitive and highly specific test. The ALK gene rearrangement could be an early mutation and it is needed as an initiating step for the carcinogenesis process. The presence of a double gene mutation, however, could be the cause of a higher-grade cancer.

Clinical utility of circulating tumor cells in patients with non-small-cell lung cancer.

Several different studies have addressed the role of the circulating tumor cells (CTC) in non-small-cell lung cancer (NSCLC). In particular, the potential of CTC analysis in the early diagnosis of NSCLC and in the prediction of the outcome of patients with early and advanced NSCLC have been explored. A major limit of these studies is that they used different techniques for CTC isolation and enumeration, they employed different thresholds to discriminate between high- and low-risk patients, and they enrolled heterogeneous and often small cohort of patients. Nevertheless, the results of many studies are concordant in indicating a correlation between high CTC count and poor prognosis in both early and advanced NSCLC. The reduction of CTC number following treatment might also represent an important indicator of sensitivity to therapy in patients with metastatic disease. Preliminary data also suggest the potential for CTC analysis in the early diagnosis of NSCLC in high-risk individuals. However, these findings need to be confirmed in large prospective trials in order to be transferred to the clinical practice. The molecular profiling of single CTC in NSCLC might provide important information on tumor biology and on the mechanisms involved in tumor dissemination and in acquired resistance to targeted therapies. In this respect, xenografts derived from CTC might represent a valuable tool to investigate these phenomena and to develop novel therapeutic strategies.

Circulating Antibodies to Linear Peptide Antigens Derived from ANXA1 and FOXP3 in Lung Cancer.

Our previous studies revealed that concentrations of circulating antibodies to annexin A1 (ANXA1) and forkhead-box P3 (FOXP3) increased significantly in patients with non-small cell lung cancer (NSCLC). This study was thus undertaken to replicate our initial findings with different sample sets. Antibodies were tested in 108 patients with NSCLC and 216 controls, who were divided into the discovery (49 vs. 108) and validation (60 vs. 108) group based on the time of enrolment. Analysis of the discovery group showed a significant increase in circulating anti-ANXA1 IgG levels in the patient group compared with the control group (p=0.005) but the validation group simply exhibited a trend toward an increase in IgG levels in NSCLC (p=0.238), generating a combined p-value of 0.009. The findings of this study support the notion that circulating IgG antibodies to ANXA1 could be used as a biomarker for early diagnosis of NSCLC but failed to replicate such findings for FOXP3.

Identification of circulating long non-coding RNAGAS5 as a potential biomarker for non-small cell lung cancer diagnosisnon-small cell lung cancer, long non-coding RNA, plasma, GAS5, biomarker.

Non-small cell lung cancer (NSCLC) is one of the most malignant cancers in the world. Early diagnosis of NSCLC has become especially important for patient treatment and prognosis. Increasing evidence suggest that long non-coding RNAGAS5 plays vital roles in cancer proliferation and differentiation in NSCLC. However, its clinical value in the diagnosis of NSCLC is unclear. The objective of this study was to evaluate the importance of circulating GAS5 as a biomarker for NSCLC diagnosis. In our study, quantitative real-time PCR(QRT-PCR) was applied to detect the GAS5 expression level in 80 pairs of cancer tissues and 57pairs of plasma samples of NSCLC patients. Further analysis was performed to study the differential expression of circulating GAS5 in 111 NSCLC patients and 78 healthy controls in our study. The results showed that GAS5 decreased in NSCLC tissues compared to noncancerous tissues (P<0.001). Furthermore, the GAS5 expression level was statistically declined in early stage of NSCLC before surgery compared with healthy controls (P<0.05) and sharply increased in postoperative groups (P=0.026). ROC curve analysis for early stage of NSCLC with the combination of GAS5, CEA and CA199 showed that the area under the ROC curve (AUC) was 0.734 (95%CI, 0.628‑0.839; P<0.0005). In conclusion, circulating GAS5 could be functioned as a potential combined biomarker for screening NSCLC and patient monitoring after surgical treatment.

Identification and characterization of miR-96, a potential biomarker of NSCLC, through bioinformatic analysis.

Lung cancer is the leading cause of cancer-related death worldwide. The poor prognosis is partly due to lack of efficient methods for early diagnosis. MicroRNAs play roles in almost all aspects of cancer biology, and can be secreted into the circulation and serve as molecular biomarkers for the early diagnosis of cancer. In the present study, we determined the expression of miR-96 and the function of its target genes in lung cancer through bioinformatic analysis. Four microRNA expression profiles of lung cancer were downloaded from Gene Expression Omnibus and the data were analyzed using SPSS16.0 software. Compared to the control group, expression of miR-96 was significantly increased in non-small cell lung cancer (NSCLC) (GSE51855), lung adenocarcinoma (GSE48414), stageI adenocarcinoma tissues (GSE63805) and the plasma of lung cancer patients (GSE68951). miR-96 was also elevated in six different NSCLC cell lines. However, the expression level of miR-96 was not related to the age, gender, clinical stage and histological subtype of the NSCLC patients. GO analysis of 78predicted target genes of miR-96 showed that 42of the obtained GO terms are highly associated with specific cellular processes including response to stimulus, signaling pathway, cell division, cell communication, cell migration and calcium signaling. KEGG results indicated that the miR-96 targets are mainly involved in the GnRH signaling pathway, long-term potentiation and insulin signaling pathway. In conclusion, miR-96, functioning as an oncogene, may play an important role in the development and progression of lung cancer. miR-96 may have the potential to serve as a molecular biomarker for the early diagnosis of NSCLC.

Significance of Serum MicroRNAs in the Auxiliary Diagnosis of Non-Small Cell Lung Cancer.

The lack of effective means for the early diagnosis of non-small cell lung cancer (NSCLC) is the leading cause of the high mortality associated with this form of lung cancer. This study aims to explore the potential significance of serum miRNA in the auxiliary diagnosis of NSCLC. The relative serum levels of 10 miRNAs in 120 patients with NSCLC, 45 patients with benign lung diseases, and 45 healthy controls were detected using real-time quantitative polymerase chain reaction (PCR). The receiver operating characteristic (ROC) curves were then used to analyze the significance of the expression of these 10 miRNAs in the diagnosis of NSCLC, as well as to compare them with the current commonly used tumor marker carcinoembryonic antigen (CEA). The serum levels of miR-125b and miR-22 in the NSCLC patients were significantly higher than those in the other two groups (p < 0.05), but the serum expression of miR-15b in the NSCLC patients was significantly lower than that in the other two groups (p < 0.01). The sensitivities of serum miR-22 and miR-15b in detecting early NSCLC (stage I + II) were significantly higher than that of CEA (p < 0.05). Area under the curves (AUCs) of serum miR-22, miR-125b, and miR-15b in the diagnosis of NSCLC were 0.725, 0.704, and 0.619, respectively, and the diagnostic significance of these three serum miRNAs for NSCLC was higher than that of serum CEA (AUC = 0.594). Serum miRNAs have potential as NSCLC-screening tumor markers, and serum miR-22 and miR15b might be used as reference indexes for the early diagnosis of NSCLC in the future.

Aberrant miR-181b-5p and miR-486-5p expression in serum and tissue of non-small cell lung cancer

BackgroundLung cancer is the leading cause of cancer deaths in China. Non-small cell lung cancer (NSCLC) is the major type of lung cancer. ObjectivesThe aim of our study was to characterize the expression profiles of miRNAs in serum and tissue of NSCLC at the same time, and to find more accurate relationship of miRNAs between serum and tissue. Furthermore, we intended to find more biomarkers of miRNAs in NSCLC samples. MethodsIn this study, the miRNAs were sequenced in 18 paired serum and 18 paired tissue samples. The expression levels of miRNAs and targets were quantified by qRT-PCR. The function analysis was performed by using bioinformatics methods. ResultsIn these paired samples miR-181b-5p was up-regulated in squamous cell carcinoma (SCC), miR-486-5p was down-regulated in adenocarcinoma (AC), and miR-21-5p was up-regulated in both SCC and AC. However, miR-181b-5p and miR-486-5p were rarely reported in lung cancer related studies. The expression levels of these two miRNAs and their targets, RASSF1 and PIK3R1 were quantified in additional samples by qRT-PCR. The results showed that the targets were negatively regulated by the two miRNAs. In addition, we noted that RASSF1 and PIK3R1 were directly involved in non-small cell lung cancer pathway. ConclusionsOur study suggested that miR-181b-5p and miR-486-5p could be new potential biomarkers for early diagnosis of NSCLC.

Identification of GlcNAcylated alpha-1-antichymotrypsin as an early biomarker in human non-small-cell lung cancer by quantitative proteomic analysis with two lectins

Background:Non-small-cell lung cancer (NSCLC) is the main type of lung cancer with high mortality rates in worldwide. There is a need to identify better biomarkers to detect NSCLC at an early stage as this will improve therapeutic effect and patient survival rates.Methods:Two lectins (AAL/AAGL and AAL2/AANL), which specifically bind to tumour-related glycan antigens, were first used to enrich serum glycoproteins from the serum of early NSCLC patients, benign lung diseases subjects and healthy individuals. The samples were investigated by using iTRAQ labelling and LC-MS/MS.Results:A total of 53 differentially expressed proteins were identified by quantitative proteomics and four glycoproteins (AACT, AGP1, CFB and HPX) were selected for further verification by western blotting. Receiver operating characteristic analysis showed AACT was the best candidate for early NSCLC diagnosis of the four proteins, with 94.1% sensitivity in distinguishing early tumour Stage (IA+IB) from tumour-free samples (healthy and benign samples, HB). The GlcNAcylated AACT was further detected by lectin-based ELISA and has better advantage in clinical application than total AACT. The GlcNAcylated AACT can effectively differentiate Stage I from HB samples with an AUC of 0.908 and 90.9% sensitivity at a specificity of 86.2%. A combination of GlcNAcylated AACT and carcinoembryonic antigen (CEA) was able to effectively differing Stage I from HB samples (AUC=0.914), which significantly improve the specificity of CEA. The combination application also has the better clinical diagnostic efficacy in distinguishing cancer (NSCLC) from HB samples than CEA or GlcNAcylated AACT used alone, and yielded an AUC of 0.817 with 93.1% specificity.Conclusions:Our findings suggest that the GlcNAcylated AACT will be a promising clinical biomarker in diagnosis of early NSCLC.

Down-expression of circulating micro ribonucleic acid (miRNA)-148/152 family in plasma samples of non-small cell lung cancer patients.

Circulating extracellular micro ribonucleic acids (miRNAs) are considered as potential biomarkers for malignancy detection and diagnosis. The aim of this study was to determine whether circulating miRNA-148/152 family (miR-148a, miR-148b, and miR-152) expression in plasma could be used as potential biomarkers for non-small cell lung cancer (NSCLC) patients and healthy individuals. The levels of miRNA-148/152 family were detected by TaqMan quantitative polymerase chain reaction (qPCR) assay in plasma of 20 NSCLC patients and 10 healthy individuals. The miRNA expression level of each sample was normalized to that of miR-16 and expressed as relative expression (2-ΔΔCt). The circulating level of all three members of miRNA-148/152 family were significantly lower in plasma samples of NSCLC patients compared with those of healthy controls (P = 0.007, P = 0.003, and P = 0.000, respectively). The expression levels of miR-148a, miR-148b, and miR-152 in the late-stage NSCLC group were all lower than the early-stage NSCLC group (all P < 0.05). The present study suggests that the expression levels of miR-148/152 family in plasma might be useful biomarkers for NSCLC patients samples in the early diagnosis of NSCLC and monitoring of tumour development.

Chromogenic Assay for Lung Cancer-Related EGFR Exon 19 Hotspot Deletion Mutations.

Epidermal growth factor receptor (EGFR) deletion mutations are associated with the development of nonsmall-cell lung cancer (NSCLC) and can serve as useful biomarkers. In the present study, a novel assay for the detection of EGFR hotspot mutations was designed to be highly sensitive and practically false-positive-free to harness the potential of detecting such mutations as biomarkers early in the diagnosis of NSCLC. The new assay draws from the polymerase chain reaction (PCR) for amplification, blue-white screening for initial allele discrimination, and Sanger sequencing for mutation confirmation. Mutant plasmids were mixed with wild-type DNA in ratios from 1:10 to 1:1000, followed by PCR amplification, blue-white screening, and sequencing. Mutants were successfully sequence confirmed for mixtures at ratios of 1:300 and 1:1000, highlighting the assay's high sensitivity and low risk of false-positives due to confirmation by Sanger sequencing. With high sensitivity and low false positives, the present assay is appealing as an aid in the early diagnosis of NSCLC through liquid biopsy. The highly customizable nature of the assay provides the possibility of applications in the early diagnosis of other cancer-related genes through nonsense-transformable mutations.

Meta-analyses of gene methylation and smoking behavior in non-small cell lung cancer patients.

Aberrant DNA methylation can be a potential genetic mechanism in non-small cell lung cancer (NSCLC). However, inconsistent findings existed among the recent association studies between cigarette smoking and gene methylation in lung cancer. The purpose of our meta-analysis was to evaluate the role of gene methylation in the smoking behavior of NSCLC patients. A total of 116 genes were obtained from 97 eligible publications in the current meta-analyses. Our results showed that 7 hypermethylated genes (including CDKN2A, RASSF1, MGMT, RARB, DAPK, WIF1 and FHIT) were significantly associated with the smoking behavior in NSCLC patients. The further population-based subgroup meta-analyses showed that the CDKN2A hypermethylation was significantly associated with cigarette smoking in Japanese, Chinese and Americans. In contrast, a significant association of RARB hypermethylation and smoking behavior was only detected in Chinese but not in Japanese. The genes with altered DNA methylation were likely to be potentially useful biomarkers in the early diagnosis of NSCLC.

Development of a multiplex methylation specific PCR suitable for (early) detection of non-small cell lung cancer

Lung cancer is a worldwide health problem and a leading cause of cancer-related deaths. Silencing of potential tumor suppressor genes (TSGs) by aberrant promoter methylation is an early event in the initiation and development of cancer. Thus, methylated cancer type-specific TSGs in DNA can serve as useful biomarkers for early cancer detection. We have now developed a “Multiplex Methylation Specific PCR” (MMSP) assay for analysis of the methylation status of multiple potential TSGs by a single PCR reaction. This method will be useful for early diagnosis and treatment outcome studies of non-small cell lung cancer (NSCLC). Genome-wide CpG methylation and expression microarrays were performed on lung cancer tissues and matched distant non-cancerous tissues from three NSCLC patients from China. Thirty-eight potential TSGs were selected and analyzed by methylation PCR on bisulfite treated DNA. On the basis of sensitivity and specificity, six marker genes, HOXA9, TBX5, PITX2, CALCA, RASSF1A, and DLEC1, were selected to establish the MMSP assay. This assay was then used to analyze lung cancer tissues and matched distant non-cancerous tissues from 70 patients with NSCLC, as well as 24 patients with benign pulmonary lesion as controls. The sensitivity of the assay was 99% (69/70). HOXA9 and TBX5 were the 2 most sensitive marker genes: 87% (61/70) and 84% (59/70), respectively. RASSF1A and DLEC1 showed the highest specificity at 99% (69/70). Using the criterion of identifying at least any two methylated marker genes, 61/70 cancer samples were positive, corresponding to a sensitivity of 87% and a specificity of 94%. Early stage I or II NSCLC could even be detected with a 100% specificity and 86% sensitivity. In conclusion, MMSP has the potential to be developed into a population-based screening tool and can be useful for early diagnosis of NSCLC. It might also be suitable for monitoring treatment outcome and recurrence.

MicroRNAs in Body Fluids as Biomarkers for Non-Small Cell Lung Cancer: A Systematic Review

Non-small cell lung cancer (NSCLC) is one of the most common life-threatening malignant tumors. A test for early diagnosis of NSCLC needs to be not too invasive and not too heavy a burden for weakened patients. A series of studies reported various microRNAs (miRNAs) could be novel serum biomarkers for NSCLC. However, the diagnostic ability of different miRNA biomarkers varies among the reports. The goal of this study was to perform a systematic review to examine the effect of miRNAs on NSCLC-related outcomes. We systematically searched The Cochrane Central Register of Controlled Trials, MEDLINE, Pub Med, EMBASE, the Chinese Biomedical Literature Database, the China Academic Journals Full-text Database, and the Chinese Scientific Journals Database for potential studies. Studies were included if they were related to miRNAs, NSCLC, and reported diagnostic outcomes. Diagnostic values analysis was used to summarize the overall test performance of miRNAs. 13 studies were included in this systematic review. The ranges of sensitivity (SEN) and specificity (SPE) of diagnosis model with miRNAs as identifying NSCLC were 0.69˜1.00 and 0.66˜1.00, respectively. The overall area under the curve (AUC) value of summary receiver operating characteristic (SROC) curve was 0.9151. The ranges of positive likelihood ratio (PLR) and negative likelihood ratio (NLR) were 2.33˜24.75 and 0.010.40, respectively. The range of diagnostic odds ratio (DOR) was 6.52˜983.38. The current evidence indicates that miRNAs in body fluids show high accuracy in identifying NSCLC, and could be a useful screening tool for diagnosing NSCLC patients.

Peripheral Blood miR-328 Expression as a Potential Biomarker for the Early Diagnosis of NSCLC

Lung cancer is often diagnosed at an advanced stage, with subsequently poor prognosis. There are no biomarkers available to facilitate early diagnosis or to discriminate between benign and malignant nodules. MicroRNAs (miRNAs) are stable molecules that can be found and measured in peripheral blood, thus representing potential diagnostic biomarkers. We evaluated 100 individuals comprising 86 patients with predominantly early-stage non-small cell lung cancer (NSCLC) and 24 healthy donors. RNA was extracted from peripheral blood samples and the expression of a panel of miRNAs was analyzed by Real-Time PCR method. Expression levels of miR-328, miR-18a, miR-339 and miR-140 were significantly higher in NSCLC patients than in healthy donors (p < 0.05). In particular, miR-328 showed good diagnostic accuracy in discriminating between patients with early NSCLC and healthy donors (AUC ROC 0.82, 95% CI 0.72–0.92), with 70% sensitivity and 83% specificity at the best relative expression cut-off of 300. Moreover, miR-339 was a good discriminant between healthy donors and late-stage NSCLC patients (AUC ROC 0.79, 95% CI 0.68–0.91). In conclusion, miR-328 represents a potential diagnostic biomarker of NSCLC, especially for the identification of early-stage tumors. Its role in discriminating between benign and malignant nodules detected by spiral CT warrants further investigation.

Identification of plasma microRNA-21 as a biomarker for early detection and chemosensitivity of non-small cell lung cancer

Studies have shown cell-free microRNA (miRNA) circulating in the serum and plasma with specific expression in cancer, indicating the potential of using miRNAs as biomarkers for cancer diagnosis and therapy. This study was to investigate whether plasma miRNA-21 (miR-21) can be used as a biomarker for the early detection of non–small cell lung cancer (NSCLC) and to explore its association with clinicopathologic features and sensitivity to platinum-based chemotherapy. We used real-time RT-PCR to investigate the expression of miR-21 in the plasma of 63 NSCLC patients and 30 healthy controls and correlated the findings with early diagnosis, pathologic parameters, and treatment. Thirty-five patients (stages NIB and IV) were evaluable for chemotherapeutic responses: 11 had partial response (PR); 24 had stable and progressive disease (SD+ PD). Plasma miR-21 was significantly higher in NSCLC patients than in age- and sex-matched controls (P < 0.001). miR-21 was related to TNM stage (P < 0.001), but not related to age, sex, smoking status, histological classification, lymph node status, and metastasis (all P > 0.05). This marker yielded a receiver operating characteristic (ROC) curve area of 0.775 (95% CI: 0.681 – 0.868) with 76.2% sensitivity and 70.0% specificity. Importantly, miR-21 plasma levels in PR samples were several folds lower than that in SD plus PD samples (P = 0.049), and were close to that in healthy controls (P = 0.130). Plasma miR-21 can serve as a circulating tumor biomarker for the early diagnosis of NSCLC and is related to the sensitivity to platinum-base chemotherapy.

Abstract 2814: Angiopoietin-like protein 3 is a novel biomarker and autoantigen in pre-diagnostic sera of non-small cell lung cancer patients

Abstract Background: Proteomic analyses of mouse lung cancer models have yielded circulating markers that can translate to human disease. Analysis of angiopoietin-like protein 3 (ANGPTL3) in pre-diagnostic human lung cancer sera identified two types of complementary, circulating markers: protein concentration and the occurrence of autoantibodies to ANGPTL3. Methods: Sera collected up to a year prior to diagnosis of lung cancer from 26 patients and 26 matched healthy controls were analyzed to determine both the circulating concentration of and autoantibody reactivity to ANGPTL3. Circulating levels were measured by commercial enzyme-linked immunosorbent assay (ELISA) and autoantibody reactivity was measured by fluorescent microarray. Additionally, sera from 38 newly diagnosed patients with lung cancer and 38 healthy individuals were also analyzed for ANGPTL3 circulating levels and autoantibody reactivity. Results: Both ANGPTL3 concentration and autoantibody reactivity were significantly elevated in sera of non-small cell lung cancer (NSCLC) patients collected 0-12 months prior to diagnosis compared to control sera (p=0.12 and p=0.035, respectively). The circulating concentration of ANGPTL3 and autoantibody reactivity were also significantly elevated in the sera of newly diagnosed NSCLC patients compared to control sera (p=0.0039 and p=0.0013, respectively). Autoantibody reactivity to ANGPTL3 and protein concentration in individual patient sera was inversely correlated. A combined analysis of autoantibody reactivity and circulating concentration improved predictive performance by receiver operating characteristic (ROC) analysis compared to individual markers, achieving 53% sensitivity at 90% specificity and an area under the curve of 81%. Western blot analysis showed expression of ANGPTL3 in NSCLC cell lines, but not in an immortalized bronchioepithelial cell line. Immunohistochemical analysis of ANGPTL3 in lung tumor tissue arrays revealed robust staining in the cytoplasm of neoplastic epithelial cells in both adenocarcinomas and squamous cell carcinomas. The addition of ANGPTL3 to a previously published panel of three autoantibodies (i.e. ANXA1, YWHAQ, and LAMR1) for early lung cancer detection improves the predictive performance of the panel to discriminate cases from controls in sera collected prior to the onset of symptoms. Conclusions: Circulating levels of ANGPTL3 and autoantibody reactivity were significantly elevated in newly diagnosed NSCLC and an independent set of pre-diagnostic samples compared to controls. A combination of circulating levels and autoantibody reactivity for ANGPTL3 improves the predictive performance. ANGPTL3 is strongly expressed in NSCLC cell lines and tissue. Our findings indicate that the occurrence of autoantibodies and the concentration of ANGPTL3 in serum may have utility for the early diagnosis of NSCLC. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2814. doi:10.1158/1538-7445.AM2011-2814

Study of distinct protein profiles for early diagnosis of NSCLC using LCM and SELDI-TOF-MS

No biomarker has been available to detect early lung cancer so far. The aim of this study is to screen biomarker patterns for early diagnosis of non-small cell lung cancer (NSCLC) using laser capture microdissection (LCM) and surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF-MS). The 3 groups of the interested cells from 13 NSCLC tissues, 11 normal lung tissues (out of the 13 NSCLC patients), and 6 benign lung diseased tissues (BLD) were successfully separated by LCM, respectively, and the homogeneities of each type of the cell populations in the three groups were estimated to be over 95%. One-hundred- and twenty-three M/Z peaks were found in the NSCLCs and normal lungs, and between the two groups the relative intensity of 98 M/Z peaks was significantly different (P < 0.05) using SELDI-TOF-MS. The diagnostic pattern constructed using support vector machine (SVM) including three proteins, M/Z 4282, 3201, and 4252 Da, respectively, showed maximum Youden Index (YI). The pattern was validated by leave-one-out cross validation (LOOCV) and the results showed that the sensitivity was 100.0%, specificity 90.9%, and positive predictive value (PPV) 92.9%. In the NSCLCs and BLDs 188 M/Z peaks were determined and 54 showed statistically difference (P < 0.05). The sensitivity, specificity, and PPV of the diagnostic pattern consisting of two proteins, M/Z 3204 and 3701 Da, were all 100.0%. So, by using LCM we have successfully purified the interested cells and solved the problem of heterogeneity of lung cancer tissue. SELDI protein chip coupled with SVM could effectively screen the differentially expressional protein profiles and eventually establish biomarker patterns with high sensitivity and specificity.

Study on detection of aberrant promoter hypermethylation of p16 and DAP kinase in serum DNA from patients with non-small cell lung cancer

To explore the feasibility of detection of aberrant methylaion of p16 and death-associated protein kinase gene as biological markers for the diagnosis of non-small cell lung cancer (NSCLC) patients. The methylation of p16 and death-associated protein kinase gene in serum and primary NSCLC tumors from 30 NSCLC patients was detected by using methylation-specific PCR methods. Aberrant methylation of at least one gene was detected in 18 of 30 (60.0%) in NSCLC tumor tissues. In these primary tumors with methylation, 9 of 18 (50.0%) samples also was detected abnormal methylation in the matched serum samples, but not in any paired normal lung tissue and healthy control subjects. Detection of aberrant methylation in the serum of NSCLC patients may have implications for early diagnosis of NSCLC.