[Clinicopathologic and prognostic significance of serum levels of cytokines in patients with advanced serous ovarian cancer prior to surgery].

OBJECTIVE: To study the clinicopathologic and prognostic significance of serum levels of six cytokines (IFN-γ, TNF-α, IL-10, IL-5, IL-4, IL-2) in patients with advanced serous ovarian cancer prior to surgery. METHODS: The serum levels of six cytokines were detected in 51 patients with advanced serous ovarian cancer and 46 healthy controls, using cytometric bead arrays. RESULTS: The serum levels of IFN-γ (20.68±11.45), IL-2 (4.54±1.18), IL-4 (5.66±2.25), IL-5 (2.72±0.86) µg/L and IL-10 (5.93±7.92) µg/L were higher (P<0.01, P<0.05) and the serum level of TNF-α (7.53±8.47) was lower (P<0.01) in patients with advanced serous ovarian cancer than those in the healthy controls. The IFN-γ/IL-4 ratio (3.93±2.34) of the patients was lower than that of the controls (P<0.01). Kaplan-Meier analysis revealed that patient's age (P=0.016), menopausal status (P=0.001) and serum IL-10 level (P=0.010) correlated significantly with patient's survival. Cox regression analysis showed that serum IL-2 (P=0.045) and IL-10 levels (P=0.007) were the independent prognostic factors. CONCLUSIONS: Patients with advanced serous ovarian cancer have Th1/Th2 imbalance and immune function disturbance. The age of patients and menopausal status are important prognostic factors. IL-2 and IL-10 level are also independent predictors of survival.

[Study of the metastasis-associated genes and its copy numbers variation in highly metastatic epithelial ovarian cancer].

OBJECTIVE: To investigate the relationship of the metastasis-associated genes and its copy numbers variation in the highly metastatic human epithelial ovarian cancer cell line HO-8910PM. METHODS: The differentially expressed genes and its copy number variation between HO-8910PM cell line and normal ovarian tissues was detected by human genome U133A 2.0 gene chip and human mapping 10K array 2.0 gene chip, and the data was analyzed by bioinformatics. Some of metastasis-associated genes were validated the results of single nucleotide polymorphism (SNP) and cDNA chips by fluorescence in situ hybridization (FISH) and real-time quantitative PCR. RESULTS: Integrate analysis of two gene chips data showed that there were 385 differentially expressed genes in the same and 379 SNP positional point (6 of them, included 2 genes) between HO-8910PM cell line and normal ovarian tissues, these copy number amplification of 379 SNP positional point of chromosome were > or = 3, which had 240, deletion < or = 1 had 139. Chromosome location analysis showed that there were 385 differentially expressed genes located at all chromosomes, and 261 of them (67.8%, 261/385) located at 10 chromosomes, included that 34 (8.8%), 33 (8.6%), 28 (7.3%), 27 (7.0%), 25 (6.5%), 24 (6.2%) of them located at chromosome 3, 2, 9, 10, 1 and 11 respectively, and 23 (6.0%) of them at chromosome 6 and 12 each, 22 (5.7%) of them at chromosome 4 and 5 each. For the function of differentially expressed genes, the results showed that 99 (25.7%) genes belonged to the family of enzymes and their regulators, 54 (14.0%) genes associated with signal transduction, 50 (13.0%) genes associated with nucleic acid binding, and 36 (9.4%) genes associated with protein binding. CONCLUSION: We have demonstrated that there are 4 kinds of differentially expressed genes related to metastasis of ovarian cancer, which belonged to the families enzyme and its regulator, nucleic acid binding, signal transduction and protein binding, and located at chromosome 1, 2, 3, 4, 5, 6, 9, 10, 11 and 12.

Identification of differentially expressed genes in the high and low metastatic human ovarian cancer cell lines and analyses of their chromosomal localizations and functions.

Oligonucleotide microarrays were used to study the differences of gene expressions in high (H) and low (L) metastatic ovarian cancer cell lines and in normal ovarian tissues (C). Bioinformatics was used to identify novel genes and their functions as well as chromosomal localizations. A total of 409 genes were differentially expressed between the high and low metastatic ovarian cancer cell lines. Of them, 271 genes were up regulated (Signal Log Ratio[SLR] > or = 1), and 138 genes were down regulated (SLR < or = -1). Except one gene whose location was unknown, all these genes were localized randomly on all the chromosomes, with a majority of them localized to Chromosomes 1, 6, 2, 17, 3, 5 and 11. Chromosome 1 contained, 43 of them (10.7%), the most for a single chromosome. A total of 264 genes (64.7%) were localized on the short arm of the chromosome (q). Functional classification showed that the 104 (25.4%) genes coding for enzymes and enzyme regulators made up the largest functional group, followed by signal transduction activity genes (43, 10.5%), nucleic acid binding activity genes (42, 10.3%), and proteins binding activity genes (34, 8.3%). These four groups accounted for 54.5% of all the differentially expressed genes. In addition, the functions of 76 genes (18.6%) were unknown. Tumor metastasis is the result of a number of genes acting in concert. The four functional groups of genes classified among these genes and their abnormalities would be the focus of further studies on ovarian cancer metastasis.

Function and chromosome location of differentially expressed genes in gastric cancer.

Using Affymetrix U133A oligonucleotide microarrays, screening was done for genes that were differentially expressed in gastric cancer (T) and normal gastric mucosa (C), and their chromosome location was characterized by bioinformatics. A total of 270 genes were found to have a difference in expression levels of more than eight times. Of them 157 were up-regulated (Signal Log Ratio [SLR] > or = 3), and 113 were down-regulated (SLR< or = -3). Except for, four genes with unknown localization, a vast majority of the genes were sporadically distributed over every chromosome. However, chromosome 1 contained the most differentially expressed genes (26 genes, or 9.8%), followed by chromosomes 11 and 19 (both 24 genes, or 9.1%). These genes were also more likely to be on the short-arm of the chromosome (q), which had 173 (65%). When these genes were classified according to their functions, it was found that most (67 genes, 24.8%) belonged to the enzymes and their regulators groups. The next group was the signal transduction genes group (43 genes, 15.9%). The rest of the top three groups were nucleic acid binding genes (17, 6.3%), transporter genes (15, 5.5%), and protein binding genes (12, 4.4%). These made up 56.9% of all the differentially expressed genes. There were also 50 genes of unknown function (18.5%). Therefore it was concluded that differentially expressed genes in gastric cancer seemed to be sporadically distributed across the genome, but most were found on chromosomes 1, 11 and 19. The five groups associated genes abnormality were important genes for further study on gastric cancer.

Gene expression profile differences in gastric cancer, pericancerous epithelium and normal gastric mucosa by gene chip.

AIM: To study the difference of gene expression in gastric cancer (T), pericancerous epithelium (P) and normal tissue of gastric mucosa (C), and to screen an associated novel gene in early gastric carcinogenesis by oligonucleotide microarray. METHODS: U133A (Affymetrix, Santa Clara, CA) gene chip was used to detect the gene expression profile difference in T, P and C, respectively. Bioinformatics was used to analyze the detected results. RESULTS: When gastric cancer was compared with normal gastric mucosa, 766 genes were found, with a difference of more than four times in expression levels. Of the 766 genes, 530 were up-regulated (Signal Log Ratio (SLR) >2), and 236 were down-regulated (SLR<-2). When pericancerous epithelium was compared with normal gastric mucosa, 64 genes were found, with a difference of more than four times in expression levels. Of the 64 genes, 50 were up-regulated (SLR>2), and 14 were down-regulated (SLR<-2). Compared with normal gastric mucosa, a total of 143 genes with a difference in expression levels (more than four times, either in cancer or in pericancerous epithelium) were found in gastric cancer (T) and pericancerous epithelium (P). Of the 143 genes, 108 were up-regulated (SLR>2), and 35 were down-regulated (SLR<-2). CONCLUSION: To apply a gene chip could find 143 genes associated with the genes of gastric cancer in pericancerous epithelium, although there were no pathological changes in the tissue slices. More interesting, six genes of pericancerous epithelium were up-regulated in comparison with genes of gastric cancer and three genes were down-regulated in comparison with genes of gastric cancer. It is suggested that these genes may be related to the carcinogenesis and development of early gastric cancer.

[Comparative proteomics analysis of differentially expressed metastasis-associated proteins in human ovarian cancer cell lines].

OBJECTIVE: To find the key proteins associated with metastasis of ovarian cancer, and find potential diagnostic markers and therapeutic targets of this malignancy. METHODS: A comparative proteomic strategy, in a combination of two-dimensional electrophoresis separation and mass spectrometry identification, was adopted to search for proteome alternations in an ovarian cancer mother cell line HO-8910 and its highly metastatic cell subline HO-8910PM. RESULTS: Twenty-one significantly different spots (two-fold increase or decrease) were detected between the two cell lines, of which 17 candidate proteins were successfully identified and characterized. Compared with those in HO-8910 mother cell line, 16 proteins were significantly up-regulated, while 5 proteins down-regulated in the highly metastatic cell subline HO-8910PM. The seventeen identified proteins could be functionally classified into 7 groups as following: zinc finger protein, calcium-binding protein, DNA repair and synthesis protein, cell regulatory protein, metabolism-related protein, cell surface antigen, cell signals and transducing protein. CONCLUSIONS: The results suggest that an obviously differential proteomic expression exists between the human ovarian cancer mother cell line HO-8910 and highly metastatic cell subline HO-8910PM. It provides a clue for further identification of metastasis-related proteins, novel diagnostic markers as well as therapeutic targets of this malignancy.

Difference of gene expression profiles between esophageal carcinoma and its pericancerous epithelium by gene chip.

AIM: To study the difference of gene expression between esophageal carcinoma and its pericancerous epithelium and to screen novel associated genes in the early stage of esophageal carcinogenesis by cDNA microarray. METHODS: Total RNA was extracted with the original single step way from esophageal carcinoma, its pericancerous epithelial tissue and normal esophageal epithelium far from the tumor. The cDNA retro-transcribed from equal quantity of mRNA was labeled with Cy5 and Cy3 fluorescence functioning as probes. The mixed probes were hybridized with two pieces of BioDoor 4 096 double dot human whole gene chip. Fluorescence signals were scanned by ScanArray 3 000 laser scanner and farther analyzed by ImaGene 3.0 software with the digital computer. RESULTS: (1) A total of 135 genes were screened out, in which 85 and 50 genes whose the gene expression levels (fluorescence intensity) in esophageal carcinoma were more than 2 times and less than 0.5 times respectively compared with the normal esophageal epithelium. (2) There were also total 31 genes, among then 27 and 4 whose expressions in pericancerous tissue were 2-fold up-regulated and 0.5-fold down-regulated respectively compared with normal esophageal epithelium. (3) There were 13 genes appeared simultaneously in both pericancerous epithelium and esophageal carcinoma, while another 18 genes existed in pericancerous epithelium only. CONCLUSION: With the parallel comparison among these three gene profiles, it was shown that (1). A total of 135 genes, Whose expression difference manifested as fluorescence intensity were more than 2 times between esophageal carcinoma and normal esophageal epithelium, were probably related to the occurrence and development of the esophageal carcinoma. (2). The 31 genes showing expression difference more than 2 times between pericancerous and normal esophageal epithelium might be relate to the promotion of esophageal pericancerosis and its progress. The present study illustrated that by using the gene chip to detect the difference of gene expression profiles might be of benefit to the gene diagnosis, treatment and prevention of esophageal carcinoma.

[Expression of a protein peak(3144 m/z) in serum and its clinical significance in patients with gastric cancer].

OBJECTIVE: To investigate the expression of protein peak (3144 m/z) in serum and of its association with clinical pathological characteristics and prognosis in patients with gastric cancer. METHODS: Three hundred and twenty seven pathologically confirmed gastric cancer patients were recruited from February 2006 to October 2008 in the Zhejiang Cancer Hospital. SELDI-TOF-MS was employed to detect the expression of protein peak (3144 m/z) in preoperative serum. RESULTS: The positive rate of 3144 m/z protein peak was 33.9% (111/327), significantly higher than that of CEA (21.1%,69/327), and the difference was statistically significant (P<0.01). The positive rate of combined detection of protein peak (3144 m/z)and CEA was 45.6% (149/327). The expression of protein peak (3144 m/z) was associated with clinical staging (P<0.01), nervous invasion (P<0.01), tumor size (P<0.01), vascular invasion (P<0.05), lymph node metastasis (P<0.05), expression of CEA (P<0.05), and depth of infiltration (P<0.05). Significant difference was observed in 3-year survival rate between the patients with protein peak and patients without protein peak (44.7% vs. 64.4%, P<0.01). However, 3144 m/z protein peak was not an independent prognostic factor on multivariate Cox regression analysis (P=0.057). CONCLUSION: Protein peak (3144 m/z) may be used as a diagnostic or prognostic marker of gastric cancer.

[Use of serum protein profiling for early diagnosis of gastric cancer].

OBJECTIVES: To identify serum biomarkers associated with early gastric cancer. METHODS: Serum proteins or peptides were purified with weak cation exchange magnetic beads in 433 patients with gastric cancer and 120 healthy subjects. Distinct peaks were selected using Biomarker Wizard software. The area under receiver operating characteristic curve(AUC) was generated to analyze discrimination capability of peaks between gastric cancers and health people. RESULTS: Thirteen distinct peaks were identified between 42 gastric cancer and 42 health people matched by age and gender(P<0.001). There were 5 peaks (2745, 2768, 6629, 3402, and 6436 m/z) with AUC greater than 0.8. Peak of 6629 m/z was identified to be transthyretin. The sensitivity and specificity of 6629 m/z were 65.5% and 92.0%. The sensitivity of 6629 m/z was 59.4% in I(A gastric cancer. CONCLUSION: Transthyretin precursor may be of value in the early diagnosis of gastric cancer.

[Application of SELDI-TOF serum proteome profiling in cervical squamous cell carcinoma].

BACKGROUND & OBJECTIVE: Up to now, there is no valid biomarker in early diagnosis of cervical cancer. Surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) is a new technique used to identify biomarkers for cancers. This study was to screen new biomarkers and build diagnostic models for early diagnosis of cervical cancer by SELDI-TOF-MS. METHODS: SELDI-TOF-MS was used to detect the serum proteomic patterns of 91 patients with early stage cervical squamous cell carcinoma, 15 patients with cervical intraepithelial neoplasia III (CIN III), and 55 healthy women (control). The serum proteomic spectra were generated on weak cation exchange (WCX2) chips. Differences in protein peaks were analyzed using Biomarker Wizard software. The diagnostic model was built by Biomarker Patterns software and further valuated by a large-scale blind test. RESULTS: A total of 122 protein peaks were detected at the molecular range of 1.5 to 20 ku, among which 19 ones were significantly different between invasive cervical squamous cell carcinomas and controls (P<0.001). A diagnostic model consisting of 2 protein peaks at 3,977 m/z and 5,807 m/z was established. Its specificity was 83.78% (31/37) and its sensitivity was 97.29% (36/37). A sensitivity of 94.44% (51/54) and a specificity of 94.44% (17/18) in a large-scale blind test were obtained. CONCLUSION: The diagnostic model consisting of 2 protein peaks at 3,977 m/z and 5,807 m/z can discriminate cervical cancer patients from healthy women.

[Screening of carcinogenesis associated genes in gastric carcinoma by gene chip].

OBJECTIVE: To screen the carcinogenesis associated genes in gastric carcinoma by gene chip. METHODS: U133A (Affymetrix Santa Clara, CA) gene chip was used to detect differentially expressed genes in tumor tissues, paratumor mucosa and normal mucosa. Bioinformatics was used to analyze the screened results. RESULTS: A total of 150 genes were detected with a difference of expression levels more than 3 times in paratumor mucosa compared with normal gastric mucosa, 130 of which were up-regulated and 20 down-regulated. According to the function classifications of the differentially expressed genes, the most common ones were enzyme and enzyme regulon activity associated genes(28, 18.7% ). The frequencies of nuclei acid binding activity associated genes,signal transduction associated genes and protein binding associated genes were 11.3%, 10%, and 8.7% respectively. Seventy-one differentially expressed genes were detected both in tumor tissues and paratumor mucosa compared with normal mucosa, 61 of which were up-regulated and 10 down-regulated. Among these 71 genes,e leven genes were localized on chromosome 19, 6 on chromosome 1, 2, 16, 17 respectively. No abnormal differentially expressed gene were detected on chromosome 5, 14, 22 and Y. CONCLUSIONS: These 71 genes differentially expressed both in tumor tissues and paratumor mucosa may be associated with carcinogenesis of gastric carcinoma. The four kinds of genes associated with enzyme and enzyme regulon activity, nuclei acid binding activity, signal transduction, and protein binding should be the main genes for the study of carcinogenesis in gastric carcinoma.

[Study on gene expression profile difference in gastric cancer, pericancerous mucosa and normal gastric mucosa from the distant cutting margin by oligonucleotide microarray].

OBJECTIVE: To study the difference of gene expression profiles in gastric cancer (T), pericancerous mucosa (P) and the gastric mucosa from distant cutting margin (C), and to screen an associated novel gene in early gastric carcinogenesis by oligonucleotide microarray. METHODS: U133A (Affymetrix, Santa Clara, CA) gene chip was used to detect the gene expression profile difference in T, P and C, respectively. Bioinformatics was used to analyze the detected results. RESULTS: When gastric cancer was compared with normal gastric mucosa, 766 genes were found,with a difference of more than four times in expression levels, including 530 up-regulated [Signal Log Ratio (SLR) > 2], and 236 down-regulated (SLR< -2). When P was compared with C, 64 genes were found, with a difference of more than four times in expression levels, including 50 up-regulated (SLR > 2), and 14 down-regulated (SLR< -2). Compared with C, a total of 143 genes with a difference of more than four times in expression levels both in T and P tissues. Of the 143 genes, 108 were up-regulated (SLR > 2), and 35 were down-regulated (SLR< -2). CONCLUSIONS: Gene chip can reveal 143 same genes both in pericancerous mucosa and gastric mucosa. These genes may be related to the carcinogenesis and development of early gastric cancer.