Surface enhanced laser desorption/ionization time-of flight mass spectrometry(SELDI-TOF-MS) is a new technique in the proteomic research. It has widely developing prospects and clinical significances. This article reviewed its technologic principium, its applications in the early diagnosis of cancer and its potential uses.

Ribosome-inactivating proteins(RIPs), a natural anti-caner protein from some advanced plants, includes single-chain robosome-inactivating proteins (type Ⅰ) and double-chain ribosome-inactivating proteins (type Ⅱ). Gelonin, relative molecular weight around 30?103, belongs to single-chain ribosome-inactivating proteins. This review is about the structure and biological activity of cell toxin Gelonin and application prospect in anti-cancer treatment.

Objective To find differential expression proteins in patients with T cell non-Hodgkin’ s lymphoma (T-NHL) by using surface-enhanced laser desorption and ionization time-of-flight mass spectrometry (SELDI-TOF-MS) technique and study their related clinical application value and prospect.Methods Serum protein of 36 T-NHL patients and 30 DLBCL patients were detected by the SELD1-TOF-MS technique and weak cation exchange (wcx-2) chip.Lactate dehydrogenase (LDH) was detected by biochemistry method.Beta2-microglobulin (β2-MG) was detected by enzyme-linked immunesorbent assay (ELISA).The significant different protein spectrometry were analyzed between DLBCL patients and T-NHL patients.The correlation analysis with protein spectrometry,disease staging,LDH and β2-MG were analyzed with Spearman.Results Nine potential candidate proteins,including the peak intensity of M/Z 1142.67,1451.43,1472.49,1512.03,3194.22,3267.41,3933.86,4593.12 and 9182.24,were identified in T-NHL patients.The 9 protein markers had no contact with disease staging of T-NHL (P ＞ 0.05).The protein markers of 4593.12 and 9182.24 were high level in T-NHL patients.LDH in these two protein markers’ positive group [(290.82±29.95) U/L,(283.94±100.94) U/L] was higher than that in negative group [(169.22±55.42) U/L,(169.50±59.25) U/L](t =-3.199,P =0.004; t =-2.378,P =0.026),and LDH was positive correlation with these two protein spectrometry (r =0.265,r =0.178,P ＜ 0.01).There was no statistically significant difference ofβ2-MG between these two protein markers’ positive group and negative group (P ＞ 0.05).The other 7 protein markers were low level in T-NHL patients,and there was no statistically significant difference of LDH and β2-MG in these 7 protein markers (P ＞ 0.05).Conclusion The protein marker of 4593.12 and 9182.24 may be the specific serological markers to identify T-NHL.The combination of these two protein markers and LDH may assess the tumor load,and provide guiding value for clinical treatment.

Objective To study the expression levels of SCCA1 and SCCA2 mRNA in tissues of cervical squamous cell carcinoma. To investigate the role of this gene in the clinical diagnosis, evaluation of treatment and observation of prognosis of cervical squamous cell carcinoma. Methods Quantitative real-time RT-PCR was used to detect the expression of SCCA1 and SCCA2 mRNA in tissues of 60 cases of cervical squamous cell carcinoma and those of 30 cases of normal cervical tissues. Results The expression level of SCCA2 mRNA in tissues of 30 cases of cervical squamous cell carcinoma was higher than in those of 15 cases of normal cervical tissues (4.405 ± 2.310, 9.088 ± 2.195) (t =-6.513, P ＜0.001), while the expression level of SCCA 1 mRNA did not significantly differ between normal and malignant tissues (P ＞0.05). The expression of SCCA2 mRNA was relevant to FIGO stages and there was a tendency for this gene to increase with the stage getting worse (F =8.313, P ＜0.05). Moreover, the overexpression of SCCA2 mRNA was significantly correlated with lymph node metastases (t =2.853, P ＜0.05). The expression of SCCA2 mRNA was not correlated with age and pathological grading (P ＞0.05). However, the expression of SCCA1 mRNA was not correlated with age,FIGO stages, lymph node metastases and histological grade (P ＞0.05). Conclusion The expression of SCCA2 mRNA may provide help for more accurate diagnosis on the clinical stages and lymph node metastases of cervical squamous cell carcinoma.

Objective To identify the biomarkers which can be used of estimating the biological behaviors of prostate cancer with osseous metastasis by SELDI-TOF-MS. Methods Screening for potential tumor biomarkers of serum samples from 19 prostate cancer patients and 35 prostate cancer patients with osseous metastasis by using the technology of SELDI-TOF-MS and CM 10 protein chips (Ciphergen Inc. USA).The PBS Ⅱ protein chip reader was used to analyze the CM10 protein chips and transform the protein information into the form of spectra. The protein contents of two groups in the same mass-to-charge ratio (M/Z value) were analyzed and preceded the analysis of variance by Biomarker Wizard software. The proteins whose contents in serum were significantly different, which was distinguished the correctly groups by Biomarker Pattern software. Results The contents of 4 proteins in the two groups were significantly different and the M/Z values of these 6 proteins were from 2000 to 20 000. The relative protein content of prostate cancer patients group was higher than that of Prostate Cancer patients with osseous metastasis group at the M/Z value of 2089,4281, 3507 and 4178 [(4.63±8.03) vs (9.88±10.77), (19.78±21.46) vs (26.73±19.41), (5.46±10.14) vs (8.10±8.74), (38.01 ±26.27) vs (45.25±20.40), (P＜0.05)]. The relative protein content of prostate cancer patients group was lower than that of prostate cancer patients with osseous metastasis group at the M/Z value of 15 900 and 16 081 [(11.52±16.80) vs (4.84±5.83), (8.55±12.64) vs (3.56±3.90), (P＜0.05)]. Conclusion The associated serum protein in prostate cancer with osseous metastasis can be quickly and exactly diagnosed by SELDI-TOF-MS with high sensitivity and specificity. This new method will be widely used in clinical application.

Objective To test serum differentially expressed proteins between early-stage (stage IB-ⅢA) and late-stage (stage Ⅳ) lung cancer patients by proteinchip technology and investigate its clinical value. Methods SELDI-TOF-MS and WCX-2 protein chip were used to detect the serum protein of 30 cases of early stage lung cancer patients and 30 cases of late stage lung cancer patients. The data were analyzed by using Biomarker Wizard software. Results There are ten different proteins in the serum between the two groups of lung cancer patients. Four protein markers 7978, 8139, 15 951 and 16 133 are over expressed and seven protein markers 2867, 6885, 8701, 8840, 13 781 and 13 955 are low expressed in the late group. Conclusion SELDI-TOF-MS proteinchip technology is a convenient, sensitive and high-throughput analysis method which can screen several relatively specific protein markers for late stage lung cancer from the serum samples. This selected protein markers can predict metastasis of lung cancer patients.

Objective To analyze the effect of miR-497 high expression on the gene expression profile of colon cancer cell line HCT116. Methods MiR-497 high expressing colon cancer cell model HCT116-497 and negative control HCT116-CON were established by lentiviral transduction. The human (V2) gene expression microarray was used to identify genes that were differentially expressed between colon cancer cells overexpressing miR-497 and the controls. The candidates were subjected to the gene ontology (GO) and KEGG pathway enrichment analysis by Molecule Annotation System 3.0 (MAS3.0). The differential expression of representative genes relative to inflammation were confirmed by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). Results Of all the differently expressed genes, 582 genes were down-regulated by at least 3-folds, which were enriched in inflammation-related signaling pathways in colon cancer cells overexpressing miR-497. The decrease in 15 representative genes was validated by qPCR. Compared with those in HCT116-CON cells, expressions of 10 genes in HCT116-497 cells, including CACNB1, FOS, IL-29, RPS6KA2, TNFSF15, IL-11, INHBC, CSF1R, JAK3 and IL-2Rβ, were decreased significantly, and there were statistical differences (all P< 0.05) Conclusion MiR-497 inhibits the mRNA expression of inflammation-related genes in colon cancer cell line HCT116.

Obiective To screen serum biomarkers in patients with hepatocellular carcinoma (HCC)by SELDI-TOF-MS technique.Methods SELDI-TOF-MS technique and CM10 Protein Chip were used to detect serum protein patterns of 46 patients with primary hepatic carcinoma and 64 healthy persons.The different proteins were obtained by the Biomarker Wizard software between the patients and healthy persons.The best biomarker of primary hepatic carcinoma was selected by evaluating the sensitivity and specificity of the protein.Results 16 protein peaks were obviously different between the patients and the healthy persons (P ＜0.05).The protein peaks of m/z 6845.70 had the highest diagnosis value with a sensitivity of 89.1％ (41/46)and specificity of 87.5 ％ (56/64).This protein was likely to be a part of the immunoglobulin heavy chain variable region.Conclusion The protein of m/z 6845.70 is potential biomarkers for the diagnosis of HCC.SELDI-TOF-MS technique is a quick,simple,convenient and high through-put technology for diagnosis of hepatocellular carcinoma.

Objective To analyze the alterations of serum protein in ESCC,compare alterations of serum protein with and without LM. Methods Serum samples were collected from 64 ESCC patients before operation and 60 cases with gender and age-matched healthy controls,special serum protein or peptide spectra was determined by SELDI-TOF-MS measurement after treating the sample onto weak cation exchange (WCX2) protein chip for each case. The serum protein profiles were compared by Biomarker Wizard Software between the ESCC patients and healthy controls, and among ESCC patients stratified according to gender, age, location of tumor, size of tumor, infiltration and with or without LM. Results (1)120 protein peaks were detected at the molecular range of 0 to 50000 in comparing of ESCC patients and healthy controls. 31 significantly different peaks were found between ESCC patients and healthy controls (P <0.05), 10 peaks were selected(P<0.01). (2) One significantly different protein peak (m/z 4174) was detected between T1 and T3, T4 (P<0.05). (3) There were three significantly different protein peaks (m/z 3970,4174 and 4277) between with LM and without LM (P<0.05).The peak (m/z 4174) was shared by two groups above. (4) No significant different protein was found when patients stratified according to gender, age, location of tumor and size of tumor. Conclusion Significant difference exists in serum proteins between ESCC patients and healthy controls. There are statistical difference exists in serum proteins between T1 and T3, T4, with LM and without LM. This difference is less than between ESCC patients and healthy controls. Some commonness is existed in serum protein fingerprint for patients with serious infiltration and with LM.

The differences in intracellular and extracellular protein expressions between human prostate cancer lines LNCap and DU145 were examined. The proteins of the two cell lines were extracted and condensed by using protein extraction kits. And the intracellular and extracellular proteins were quantitatively detected on a micro-plate reader by using bicinchoninic acid (BCA) method. The proteins in cell culture fluid were qualitatively assayed by SELDI-TOF-MS. The results showed that the intracellular protein contents of LNCap cells were extremely higher than those of DU145 cells. After serum-free culture, both intracellular and extracellular protein contents of LNCap and DU145 were decreased to some extent. And the intracellular proteins were decreased by 5% in LNCap and by 36% in DU145 respectively, while the extracellular proteins were decreased by 89% in LNCap and 96% in DU145 respectively. SELDI assay revealed that there were 5 marker proteins in LNCap and 6 in DU145. Although both LNCap and DU145 cell lines originated from human prostate cancer, they had some differences in protein expression.
Cell Line, Tumor ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Mass Spectrometry/*methods ; Prostatic Neoplasms/*metabolism ; Proteins/chemistry ; Proteins/metabolism ; Proteomics/*methods ; Tumor Markers, Biological