Application of surface-enhanced laser desorption/ionization time-of-flight-based serum proteomic array technique for the early diagnosis of retinoblastoma / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To find new biomarkers in the sera of retinoblastoma (Rb) patients with surface enhanced laser desorption/ionization time of flight mass spectrometry (SELDI TOF MS) and protein chip technique.METHODS:SELDI TOF MS, IMAC30 and CM10 protein chips were used to analyze the protein profiles from sera of 18 patients with Rb and 17 age matched controls. The protein profiling was analyzed statistically by Ciphergen protein chip software 3.0.2. The test was applied to compare the protein peak intensity. Fisher's exact test was used to compare the predominance of differential protein peaks appeared in patients.RESULTS:With IMAC30 protein chips, there were 26 proteins which appeared different in sera of patients with Rb compared to normal children. Among them, 21 proteins, I.e. 7746, 7014, 11713, 3049, 7084, 7299, 5888, 2544, 12575, 5489, 9658, 9575, 9929, 10161, 8955, 1886, 10617, 6209, 2411, 7374, 6614m/z were up regulated and 5 proteins, I.e. 8382, 7923, 7972, 8590, 66576m/z, were down regulated(P<0.01). Using the 7014 protein peak for statistical analysis, we could differentiate the patients with Rb from the healthy children with a sensitivity of 94.4% and a specificity of 82.4%. By CM10 protein chips, 4 proteins, including 3 up regulated proteins(5888, 6097, 7798 and 1 down regulated protein (8590m/z), were detected in Rb patients (P<0.01). The sensitivity and specificity were 83.3% and 70.6% respectively when 7798m/z protein peak was selected for statistical analysis.CONCLUSION:There are a few candidates as Rb biomarkers in the sera of Rb patients. SELDI TOF MS protein chip technology could be a potential method in the clinical screening test of Rb.

Protein profile of human lung squamous carcinoma cell line NCI-H226 / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To construct a database of human lung squamous carcinoma cell line NCI-H226 and to facilitate discovery of novel subtypes markers of lung cancer.</p><p><b>METHOD</b>Proteomic technique was used to analyze human lung squamous carcinoma cell line NCI-H226. The proteins of the NCI-H226 cells were separated by two-dimensional gel electrophoresis and identified by mass spectrometry.</p><p><b>RESULTS</b>The results showed that a good reproducibility of the 2-D gel pattern was attained. The position deviation of matched spots among three 2-D gels was 1.95 +/- 0.53 mm in the isoelectric focusing direction, and 1.73 +/- 0.45 mm in the sodium dodecyl sulfate-polyacrylamide gel electrophoresis direction. One hundred and twenty-seven proteins, including enzymes, signal transduction proteins, structure proteins, transport proteins, etc. were characterized, of which, 29 identified proteins in NCI-H226 cells were reported for the first time to be involved in lung cancer carcinogenesis.</p><p><b>CONCLUSION</b>The information obtained from this study could provide some valuable clues for further study on the carcinogenetic mechanism of different types of lung cancer, and may help us to discover some potential subtype-specific biomarkers of lung cancer.</p>

Serum amyloid A protein: a potential biomarker correlated with clinical stage of lung cancer / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To identify serum diagnosis or progression biomarkers in patients with lung cancer using protein chip profiling analysis.</p><p><b>METHOD</b>Profiling analysis was performed on 450 sera collected from 213 patients with lung cancer, 19 with pneumonia, 16 with pulmonary tuberculosis, 65 with laryngeal carcinoma, 55 with laryngopharyngeal carcinoma patients, and 82 normal individuals. A new strategy was developed to identify the biomarkers on chip by trypsin pre-digestion.</p><p><b>RESULTS</b>Profiling analysis demonstrated that an 11.6 kDa protein was significantly elevated in lung cancer patients, compared with the control groups (P < 0.001). The level and percentage of 11.6 kDa protein progressively increased with the clinical stages I-IV and were also higher in patients with squamous cell carcinoma than in other subtypes. This biomarker could be decreased after operation or chemotherapy. On the other hand, 11.6 kDa protein was also increased in 50% benign diseases of lung and 13% of other cancer controls. After trypsin pre-digestion, a set of new peptide biomarkers was noticed to appear only in the samples containing a 11.6 kDa peak. Further identification showed that 2177 Da was a fragment of serum amyloid A (SAA, MW 11.6 kDa). Two of the new peaks, 1550 Da and 1611 Da, were defined from the same protein by database searching. This result was further confirmed by partial purification of 11.6 kDa protein and MS analysis.</p><p><b>CONCLUSION</b>SAA is a useful biomarker to monitor the progression of lung cancer and can directly identify some biomarkers on chip.</p>

Application of surface-enhanced laser desorption/ionization time-of-flight-based serum proteomic array technique for the early diagnosis of prostate cancer / 亚洲男科学杂志(英文版)

<p><b>AIM</b>To identify the serum biomarkers of prostate cancer (PCa) by protein chip and bioinformatics.</p><p><b>METHODS</b>Serum samples from 83 PCa patients and 95 healthy men were taken from a mass screening in Changchun, China. Protein profiling was carried out using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS). The data of spectra were analyzed using two bioinformatics tools.</p><p><b>RESULTS</b>Eighteen serum differential proteins were identified in the PCa group compared with the control group (P < 0.01). There were four proteins at the higher serum level and 14 proteins at the lower serum level in the PCa group. A decision tree classification algorithm that used an eight-protein mass pattern was developed to correctly classify the samples. A sensitivity of 92.0% and a specificity of 96.7% for the study group were obtained by comparing the PCa and control groups.</p><p><b>CONCLUSION</b>We identified new serum biomarkers of PCa. SELDI-TOF MS coupled with a decision tree classification algorithm will provide a highly accurate and innovative approach for the early diagnosis of PCa.</p>

A preliminary analysis of non-small cell lung cancer biomarkers in serum / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To identify potential serum biomarkers that could be used to discriminate lung cancers from normal.</p><p><b>METHODS</b>Proteomic spectra of twenty-eight serum samples from patients with non-small cell lung cancer and twelve from normal individuals were generated by SELDI (Surfaced Enhanced Laser Desorption/Ionization) Mass Spectrometry. Anion-exchange columns were used to fractionate the sera into 6 designated pH groups. Two different types of protein chip arrays, IMAC-Cu and WCX2, were employed. Samples were examined in PBSII Protein Chip Reader (Ciphergen Biosystem Inc) and the discriminatory profiling between cancer and normal samples was analyzed with Biomarker Pattern software.</p><p><b>RESULTS</b>Five distinct potential lung cancer biomarkers with higher sensitivity and specificity were found, with four common biomarkers in both IMAC-Cu and WCX2 chip; the remaining biomarker occurred only in WCX2 chip. Two biomarkers were up-regulated while three biomarkers were down-regulated in the serum samples from patients with non-small cell lung cancer. The sensitivities provided by the individual biomarkers were 75%-96.43% and specificities were 75%-100%.</p><p><b>CONCLUSIONS</b>The preliminary results suggest that serum is a capable resource for detecting specific non-small cell lung cancer biomarkers. SELDI mass spectrometry is a useful tool for the detection and identification of new potential biomarker of non-small cell lung cancer in serum.</p>