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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 UI33A 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 ≥3, which had 240, deletion ≤ 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.
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Objective To detect the level of Tnl and T.2 type cytokines in the patients with lym-phoma in order to find out the laboratory evidence of tumor immunotherapy. Methods The levels of serum IFN-γ, TNF-α, IL-2, IL-4, IL-5 and IL-10 were measured by cytometric bead array (CBA) in 92 patients with lymphoma, and 70 normal sera as control. Results The levels of TH1 type cytokincs in 92 patients with lymphoma were: IFN-γ (34.26 ± 33.48) pg/ml, TNF-α (8.17 ± 10. 09) pg/ml, IL-2 (3.74 ±1.72) pg/ml; and the levels of TH2 type cytokines were: IL-10 (6. 28±8.56) pg/ml, IL-5 (3.53 ±3.20) pg/ml, IL-4 (6.22±7.13) pg/ml. The levels of TH1 and TH2 cytokines in lymphoma patients were significantly higher than those in controls(P ＜ 0.01) except TNF-α. And the rate of IL-2/IL-4 was signifi-cantly decreased in lymphoma patients(P ＜0.01). The level of IL-10 in Ⅲ/Ⅳ stage lymphoma patients was much higher than that in Ⅰ/Ⅱ stage patients(P ＜0.01). The level of IFN-γ/was significantly decreased in aged patients with lymphoma (P ＜0. 05). Conclusion TH 1/TH2 is imbalance in lymphoma, which may provide clinical index for the evaluation of progrossien and prognosis. In the lymphoma patients TH1/TH2 shifts to TH2, which may be the mechanism of tumor arising and transferring by immune escape from immu-nosurveillance.
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Objective To analyze the serum proteomic pattern of the cervical cancer patients,to develope diagnostic model and to evaluate its clinical significance.Methods WCX magnetic bead and MALDI-TOF were used to detect the serum proteomic pattern of 77 patients with cervical squanmous cell carcinomas,13 patients with CINⅢ and 52 healthy women.Biomarker Wizard software was used to detect protein peaks and potential difference between cervical cancer and controls.The model was developed by Biomarker Patterns software.Results A diagnostic pattern consisting of three differential protein peaks was established with 100%(32/32)sensitivity and 93.8%(30/32)specificity.A sensitivity of 77.8%(35/45)and a specificity of 75%(15/20)in blind test were obtained.The diagnostic model also could discriminate CINⅢ and SCC-Ag negative patients from controls.Conclusion The diagnostic pattern combining 3974,3398,13732m/z protein peaks can discriminate not only cervical squamous cell cancer but also CINⅢ and SCC-Ag negative patients from controls.
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<p><b>OBJECTIVES</b>To study the difference between gene expressions of high (H0-8910PM) and low (HO-8910) metastatic human ovarian carcinoma cell lines and screen novel associated genes by cDNA microarray.</p><p><b>METHODS</b>cDNA retro-transcribed from equal quantities of mRNA derived from high and low metastatic tumor cells or normal ovarian tissues were labeled with Cy5 and Cy3 fluorescein as probes. The mixed probe was hybridized with two pieces of BioDoor 4096 double dot human whole gene chip and scanned with a ScanArray 3000 laser scanner. The acquired image was analyzed by ImaGene 3.0 software.</p><p><b>RESULTS</b>A total of 355 genes with expression levels more than 3 times larger were found by comparing the HO-8910 cell with normal ovarian epithelial cells. A total of 323 genes with expression levels more than 3 times larger in HO-8910PM cells compared to normal ovarian epithelium cells were also detected. A total of 165 genes whose expression levels were more than two times those of HO-8910PM cells compared to their mother cell line (HO-8910) were detected. Twenty-one genes with expression levels > 3 times were found from comparison of these two tumor cell lines.</p><p><b>CONCLUSIONS</b>cDNA microarray techniques are effective in screening differential gene expression between two human ovarian cancer cell lines (H0-8910PM; HO-8910) and normal ovarian epithelial cells. These genes may be related to the genesis and development of ovarian carcinoma. Analysis of the human ovarian cancer gene expression profile with cDNA microarray may help in gene diagnosis, treatment and prevention.</p>
Subject(s)
Female , Humans , Gene Expression Profiling , Neoplasm Metastasis , Oligonucleotide Array Sequence Analysis , Ovarian Neoplasms , Genetics , Pathology , Tumor Cells, CulturedABSTRACT
three years survival group,and those who died within 3 yrs had P-gp positive expression(24 cases,53.3%).Comparing of the two groups showed statistical significance(P3 year survival stage group,and the patients who died within three years group,the P27 expression was positive in 26 cases(56.5%)(P
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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.
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Objective: To study the propagation and phenotypes changes of killer cell (CD3AK cell activated by CD3 mAb in vitro. Methods: Lymph nodes taken from lung cancer patient is dissociated into single cell suspension by mechanical method and cultured in culture medium added CD3 mAb and a little dose IL-2. We analyze cell immunophenotype by flow cytometry and proliferation by trypan blue exclusion test per 2 days. Results: Immunophenotypic analysis showed that CD3AK expressing CD3, CD8, CD56, CD25 increased, and reached a peak value which is 2.33 times than before culturing in the 8 th day. Conclusion: CD3 mAb added to the culture medium can obviously activate CD3AK cell and stimulate proliferation and keep its killer activity.