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Objective To identify the changes of DNA methylation profile in the process of malignant transformation of BEP2D cell induced by α particles.Methods The genomic DNAs were isolated from the malignant transformation BERP35T4 cells and immortalized human bronchial epithelial cell line BEP2D.Genomic DNAs were digested by MseI and ligated of PCR linkers.Methylated DNAs were digested by BstUI and amplified by PCR.The methylated DNA probes were prepared by labeling with Cy3 and Cy5 fluorescence dyes individually and hybridized to the methylation CpG-Island microarray.The hybridization results were scanned and analyzed.Intensity values were quality controlled and normalized.The normalized data were used to identify the differentially expressed genes based on a 1.5 fold difference of the expression level.Results There were 16 genes which showed changes of methylation level in malignant transformation BERP35T4 cells, 9 of them were hypermethylation and 7 were hypomethylation.These genes were including the SKIP gene, PPP3CC gene, MAP2K6 gene, KIR2DL1 gene, KIR2DL4 gene, KIR3DP1 gene, ZNF493 gene, ZNF100 gene, NKX2-5 gene, TFAP2D gene, DR1 gene, KCNJ16 gene, CCDC18 gene, FNBP1L gene, IRX4 gene, EPB41L3 gene, TCP10 gene and so on.Conclusions The DNA methylation might have effects on ionizing radiation drived tumorigenesis.
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<p><b>BACKGROUND</b>To screen and identify differentially expressed genes among lung cancer tissues, paracancerous pulmonary tissues and some other kinds of tumor tissues using suppression subtractive hybridization (SSH) and cDNA Microarray.</p><p><b>METHODS</b>One cDNA chip was made by gathering clones of three differentially expressed cDNA libraries which came from BEP2D cell lines during three different malignant transformed phases. Then the clones were hybridizated with cDNA probes which extracted from 15 cases of lung cancer tissues, 5 cases of paracancerous pulmonary tissues and 24 cases of other 8 kinds of tumor tissues respectively.</p><p><b>RESULTS</b>Twenty-six cDNAs were obtained which expressed higher in lung cancer tissues than that in paracancerous pulmonary tissues. Thirty-one cDNAs expressed remarkably higher in paracancerous tissues than those in cancer tissues. Compared with other 8 kinds of tumors, paracancerous tissues had 63 overexpressed cDNAs and lung cancer tissues had 87 overexpressed cDNAs.</p><p><b>CONCLUSIONS</b>The combination of SSH and cDNA microarray is rapid and effective for screening and identification of differentially expressed genes in different samples. It may be potentially useful for diagnosis of lung cancer to further study the differentially expressed genes among lung cancer tissues, paracancerous pulmonary tissues and other tumor tissues.</p>
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<p><b>BACKGROUND</b>To profile the expression patterns of 60 lung cancer related genes in human bronchial epithelial cell (BEP2D) and alpha-particle induced malignantly transformed cell (R15Hp35T-2).</p><p><b>METHODS</b>Sixty lung cancer related cDNAs were micro-arrayed onto the microscope slides using Cartesian PixSys5500 cDNA Microarray machine. Total RNA from BEP2D cell and R15Hp35T-2 cell was extracted and labeled by fluorescent dye. The labeled probe was then hybridized with the cDNA.</p><p><b>RESULTS</b>Compared with the BEP2D cell, 27 genes up-regulated and 7 down-regulated in the R15Hp35T-2 cell. The expression abundance of most tumor suppressor genes were similar in the two kinds of cells, however, most oncogenes and growth factor genes were overexpressed in R15Hp35T-2 cell.</p><p><b>CONCLUSIONS</b>In malignantly transformed human bronchial epithelial cell model induced by alpha-particle, some oncogenes and growth factor genes may promote the malignant transformation together.</p>
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Objective:To isolate genes related to benflumetol resistance in the rodent malaria P.berghei K173 strain.Methods:The mRNA differential display technique was used to identify differentially expressed genes between drug-sensitive P.berghei K173 strain and benflumetol-resistant strain selected under benflumetol pressure.Northern blot was performed in order to determine the genes actually derived from maternal strains.Results:PCR reactions were prepared respectively with twenty-four pairs of primers in each of the strains.Thirty-three differentially displayed genes fragments were cloned and eight of them were sequenced. The eight sequences were novel ESTs and were submitted to GenBank. Homology searching revealed that clone CB52 was slightly homologous to P.falciparum cyclophilin gene. Hybridization confirmed that the gene fragment CB52 was upregulated in the benflumetol-resistant parasite strain, while downregulated in the chloroquine-resistant strain.Conclusions:CB52 fragment was involved in the benflumetol resistance in P.berghei and the genetic mechanisms of drug-resistance to benflumetol and chloroquine were different.
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Objective To study protein variation between PtenL/LMEFs and Pten△/△MEFs cells.Methods Two-dimensional electrophoresis(2-DE) was employed to compare the differential expression proteins between PtenL/LMEFs and Pten△/△MEFs cells.Six differential expression proteins were digested in gel by enzyme and the mass of generated peptides was measured by matrix assisted laser desorption ionization time of flight mass spectrometry(MALDI-TOF-MS).The data obtained from peptide mass fingerprinting(PMF) were searched using the Internet available database and five proteins were identified.Results Compared with that of PtenL/LMEFs,expression level of proteins including phosphoglycerate mutase 1(PGAM1) and peptidyl-prolyl cis-trans isomerase C(PPIC) was up-regulated,whereas expression level of Transgelin 2 was down-regulated in Pten△/△MEFs cells.B and F proteins were both identified to be peroxiredoxin-6.They had similar molecular weight but different PI which might be caused by post-translation modification.B protein was only expressed in Pten△/△MEFs cells.Conclusion The protein profile of Pten△/△MEFs cells displayed obvious difference compared to that of PtenL/LMEFs cells.The results implied that various distinct different proteins might lead to cancer.
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The bioavailability of zinc in normal and zinc deficient LACA mice was investigated using isotope zinc-65. The intestinal absorption of Zn in the zinc-deficient mice (71.913.6%) was significantly higher than that in the normal mice (38.8?6.8%). No difference on Zn distribution in the body was observed. Specific activity of zinc-65 in liver, spleen, kindey and muscle in the zinc-deficient mice was significantly higher than that in the normal mice. The retention curve of zinc-65 in mice was properly fitted into two exponential functions. The biological half-times of zinc in blood, spleen, lung, femur and muscle attached to femur in the zinc-deficient mice was slower and ration ratio in the slower clearance component in the zinc-deficient mice, was larger than that in the normal mice.