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Objective@#Vascular endothelial growth factor A (VEGFA) was investigated as the key protein which might promote the specific metastasis progress of nasopharyngeal carcinoma.@*Methods@#Sixteen specimens of pulmonary metastasis carcinoma and counterparts in primary nasopharyngeal carcinoma tissue were collected from patients. The expression of VEGFA through immunohistochemistry was investigated.VEGFA was knocked down by siRNA in two cell lines of nasopharyngeal carcinoma (CNE-1 and 5-8F), MTT and Transwell test were used to explore the role of VEGFA in praxiology. Then shRNA was used to cultivate the stable CNE-1 cell line with down-regulated-expression of VEGFA. The nude mice models were built through tail vein injection of specific nasopharyngeal carcinoma cells, and lungs were collected to perform further metastasis analysis.@*Results@#Previous genetic studies showed that VEGFA had higher expression in metastasis tissue, and the result was validated in the present study using immunohistochemistry. The percentage of positive cells was 84.8% in pulmonary metastasis group, 51.5% in primary tissue group (t=8.639, P<0.05), average optical density was 0.154 in pulmonary metastasis group, 0.061 in primary tissue group (t=18.791, P<0.05). Low expression of VEGFA inhibited cell viability of optical density value of CNE-1 in siRNA gourp was 0.715, 0.902 in control group (t=7.274, P<0.05); 5-8F in siRNA group was 0.715, 0.935 in control group (t=7.751, P<0.05). Number counting suppressed migration of CNE-1 in siRNA group was 52 per high-power lens, 124 per high-power lens in control group (t=29.380, P<0.05), 5-8F in siRNA group was 65 per high-power lens, 155 per high-power lens in control group (t=18.181, P<0.05). Number counting invasion of CNE-1 in siRNA gourp was 38 per high-power lens, 86 per high-power lens in control group (t=27.665, P<0.05), 5-8F in siRNA group was 52 per high-power lens, 116 per high-power lens in control group (t=40.972, P<0.05) in vitro. Furthermore, knock-down of VEGFA in nasopharyngeal carcinoma reduced the pulmonary metastasis in vivo. Number counting of tumor volumes in shRNA group was 2.4, and 11.0 in control group (t=6.143, P<0.05); average optical density of immunohistochemistry in shRNA group was 0.033, and 0.176 in control group (t=15.734, P<0.05).@*Conclusions@#Results above reveal the overexpression of VEGFA in nasopharyngeal carcinoma can facilitate the pulmonary metastasis. Targeting VEGFA may provide a new biomarker of clinical study.
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In recent years,with the completion of the Human Genome Project and the development of mapping the Human Ge?nome Methylation Variable Site Map Plan,research on epigenetics and the generation and deveopment of cancer,epigenetic treatment drugs,especially the successful clinical application of the DNA methyltransferase and histone deacetylation inhibitors in the treatment of cancer patients,epigenetic has becoming a hot spot. This article reviews the recent progress in pharmacological action of epigenetics-based anticancer drugs,it may provide some new ideas to the therapy and fundamental research of cancer.
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Polyunsaturated fatty acids (PUFAs) are important structural fatty acids of biological membrane. They play important roles in regulation of lipid metabolism,stimulation of growth and development,protection a gainst cancer,retardation of aging,immuno-regulation, cardiovascular health,and bodymass loss. In recent years, researchers have found t hat ω-6 PUFAs can promote tumor angiogenesis,while ω-3 PUFAs have the properties of inhibiting tumor angiogenesis. This review focuses on the effects of these two types of fatty acids on tumors,especially on the regulation of tumor angiogenesis.
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Objective To analyze the differential gene expression profiling of liver in rats subjected to hemorrhagic shock(HS) and sham hemorrhage shock(SHAM) by gene chip technology, thus to evaluate the possible molecular pathogenesis of HS. Method 20 male Wistar rats were randomly divided into a SHAM group and a HS group, with 10 rats in each group. Hepatic gene expression profiles were detected by oligonucleotide microarrays of 5705 mouse genes in two groups for three times. Genes with ratio(R) > 2 were identified as up-regulated and R < 0.5 were identified as down-regulated. Biological function of differentially expressed genes was analyzed and 9 genes were selected to undergo semi-quantitative RT-PCR. Results Among the total 5705 probes detected,86 genes showed differential expression in HS group comparison with SHAM group. The expression levels of 72 genes were up-regulated while those of 14 genes were down-regulated significantly. Differentially expressed genes were classified according to their biological function: transport genes, transcription regulator genes, signaling genes, response to stress genes, metabolic genes, development genes and cell adhesion genes. Conclusions cDNA microarray is an efficient and high-throughout method to survey gene expression profiles in HS.The variation of those gene expressions might be a potential pathogenic mechanism for HS that may offer a novel target for further study of therapeutic strategies of HS.
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Objective Human immunoglobulin combinatorial library was generated by using phage surface display expression system, and phage antibodies (Fabs) to platelet were screened.Methods:PBMC were separated from a patient who Were transfusion refractory to platelet. mRNA was isolated and cDNA was synthesized by reverse transcriptase.The immunoglobulin heavy chain Fd and the light chain ? genes were amplified by half nested PCR and the PCR products were cloned into the expression vector pCome3 respectively.The immunoglobulin combinatorial library was constructed and screened by 3 rounds of affinity selection.Results Human Immunoglobulin Combinatorial Library was successfully constructed.The specific phage antibodies were highly enriched after 3 rounds of biopanning selection against platelet membrane proteins.Conclusion The antibody library and human monoclonal antibodies to platelet may be useful as molecular tools to study the anti platelet drugs, platelet related diseases, epitope of human platelet antigens.
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Objective To isolate and purify the membrane protein of SeAx cells, which was derived from cutaneous T-cell lymphoma cell line (CTCL), and to prepare polyclonal antibodies against membrane proteins. CTCL antigens were screened with SEREX method from the cDNA expression library of SeAx cells. Methods SeAx cells were cultured to 2?1010 and membrane proteins were purified by means of differential centrifugation after homogenate was made, and organelle protein specific antibodies were used to perform Western blot. Rabbit was immunized 4 times by the membrane proteins, and the serum antibody titer was measured by ELISA after each immunization. The specific interactions between antibodies in serum and SeAx proteins were detected by Western blotting. E. coli were initially transfected by recombinant phages, and the recombinant proteins expressed were transferred onto the nitrocellulose membranes, which were then again reacted with diluted serum from the immunized rabbit. Positive clones were subcloned and the nucleotide sequence of the cDNA inserts was determined. Finally, cDNA sequence analysis via database searching was performed to define the gene. Results Membrane proteins of SeAx cells were purified by differential centrifugation and a 1?10-5 titer of antibody was obtained after 4 times of immunization. 1?106 phage clones were screened, 25 of which were positive. Nucleotide sequencing and database searching showed that 4 cDNA inserts were cell membrane proteins, respectively as integrin alpha 4,ligatin, matrix metalloproteinase 24 and MHC-I molecule HLA-A. Conclusion Membrane proteins of CTCL and SeAx cells, are purified and polyclonal antibodies against SeAx cell membrane proteins are successfully prepared. The 4 genes of membrane proteins are identified by means of SEREX from SeAx cDNA library, expressed by phages. These genic candidates are able to induce systemic humoral immune reactions and might therefore be promising targets for immunotherapeutic interventions of CTCL.