Screening Differential Expression of Taxol-resistance Related Gene of Ovarian Carcinnoma by cDNA Microarray
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Objective To study the gene differential expression of taxol resistance cell line OC3/Tax300 of ovarian carcinnoma compared with other parent cell OC3,screen drug-resistance related gene by cDNA microarray, and discuss the reiationship between gene expression difference and drug-resistance.Methods The cDNA retro-transcribed from equal quantity mRNA derived from OC3/Tax300 and OC3 which were labeled with Cy5 and Cy3 fluorescence as probe respectively. The mixed probe were hybridized with BiostarH140S gene expression chip. The acquired image was analyzed by software.Results 134 signitificantly differently expressed genes were screened out, of which up-and down regulated genes were 17 and 117 respectively. They were related to 14 kinds of genes. The down-regulated genes were EBNA-3,COP9, StIP1,and so on. The up-regulated genes were JAK2,HSPS,NADH, and so on.Conclusion These genes related to the mechnisms of drug-resistance of ovarian cancer.Keywords:
Differential display
Gene chip analysis
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Objective To study the gene expression profiles of ovarian cancer by cDNA microarray,and preliminarily analyze the function of part of differential expression genes.Methods BiostarH-40s gene microarray containing 4 097 genes was used to analyze gene expression patterns in tissue samples from 5 cases of human ovarian serous cystadenocarinoma(cy5-dUTP present ) and 5 cases of normal ovarian tissues(cy3-dUTP present).Results 163 genes of differential expression were found in more than 4 cases of ovarian cancer,the expression of 66 genes increased(up-regulated),the expression of 97 genes decreased(down-regulated). 37 differential genes with difinite gene function classification were found including three protocogene and tumor suppressor genes,one cyclin protein gene,three cytoskeletal and movement protein genes,one DNA binding,transcription and transcription factors gene,two cell receptor genes,five immune-related protein genes,seven metabolism genes,two protein translation and synthesis genes,three growth-related genes and seven other types of genes.Conclusion Gene expression profiles of ovarian cancer can be detected by cDNA microarray,and differential expression genes and their gene function classification of ovarian cancer are found.
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Objective To screen lung cancer me ta stasis-associated genes by analysis of differentially expressed genes between human lung giant cell carcinoma cell strains of high metastatic 95D and low meta static 95C using cDNA microarray.Methods The mRNAs were extracted from human lung giant cell car cinoma cell strains of 95D and 95C,and reversely transcribed to the cDNAs,w hich were then labeled by Cy5-dUTP and Cy3-dUTP to prepare the hybridization p robes.The mixed probes were hybridized with the cDNA microarray.The figure was obtained by scanning the cDNA microarray chip with Scan Array 3000 scanner and the information was managed with ImaGene3.0 software.Thus the genes chip of dif ferent expression between high metastatic 95D and low metastatic 95C was screen ed.A part of genes whose expression was changed in cDNA microarray analysis wer e further identified by RT-PCR.Results The cDNA microarray analysis showed that the expression of 466 genes was changed.Among them,108 pairs of double gene had the same Gen bank ID,including 47 pairs of up-regulation genes and 61 pairs of down-regula tion genes.Fifty-eight genes were not elsewhere classified.Some of the genes,including KIAA1108,PGR1,JWA,S182,Jab1 etc,were further confirmed by RT-PC R.The results of RT-PCR coincided well with the cDNA microarray results.Conclusion Many different genes are involved in the metastasis of lung cancer.cDNA microarray technique might be effective in screening lung canc er metastasis-associated genes.
Gene chip analysis
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Objective To screen the human pancreatic cancer-associated genes by cDNA microarray.Methods The PCR products of 14 000 genes were spotted onto a chemical-material-coated-glass plates in array.DNAs were fixed onto the glass plate after series of treatments.The total RNAs were isolated from 4 specimens of the pancreatic cancer and the normal tumor-surrounding pancreatic tissues.Both equal quantity RNAs were reversely transcribed to cDNAs and labeled with the fluorescent Cy5-dCTP and Cy3-dCTP to prepare the hybridization probes.The mixed probes were hybridized to the cDNA microarray.After high-stringent washing,the fluorescent signals were scanned by ScanArray 4000 scanner.The values of Cy5-dCTP and Cy3-dCTP on each spot were analyzed and calculated by GenePix Pro 3.0 software.Results By applying this cDNA microarray,189 differentially expressed genes in pancreatic cancer,whose ratios of Cy5/Cy3 were higher than 2.0 or lower than 0.5,were screened out among the 14 000 target genes,comprising 101 known genes and 88 novel gene.Among the known genes,upregulated and downregulated genes were 50 and 51 respectively,including oncogenes and tumor suppression genes,cell cycle related genes,cell skeleton and cinetc-protein related genes,cell apoptosis related genes,DNA synthesis related genes,DNA damage and repair-related genes,transcription factors,receptor related genes,immune related genes,signal transduction related genes,protein translation and synthesis related genes,metabolism related genes.Conclusion The analysis of gene expression profile of tumor based on cDNA microarray can realize high-throughput screening of the genes associated with the pancreatic cancer,and it can help to rapidly explore the genes function.Further analysis of the obtained genes will help to understand the molecular mechanism of the pancreatic cancer.
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Objective:To screen differential expression of Gene in ovarian cancer by eDNA microarray,and preliminarily analyze the function of these genes.Methods:We used eDNAmicroarray which contain 4097 genes to analyze gene expression pattern in tissue samples from 4 cases human ovarian cancer and normal ovarian tissue to screen genes of differential expression.Results:163 Genes of differential expression were detected in ovarian cancer,expression of 66 genes increased(up-regulated),the function of 31 genes have been acknowledged in 66 genes(up-regulated expression),expression of 97 genes decreased(down-regulated).The function of 51 genes have been acknowledged in 97genes(down-regulated expression).Conclusion:Genes of abnormal expression in ovary cancer can be detected by eDNA microarray,it afford reference to study these genes deeply.
Gene chip analysis
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Objective To screen for the differentially expressed genes in breast cancer cell line MCF 7 and tamoxifen resistant breast cancer cell line LCC2 by using cDNA microarray Methods The PCR products of 8000 genes were spotted on chemical material coated glass plates in array The DNAs were then fixed on the glass plate by a serial of treatments The total RNAs were isolated from cells cultured in the flash, and then were purified to mRNA by Oligotex Both the mRNAs from different breast cancer cell lines were reversely transcribed to cDNAs with the incorporations of fluorescent dUTP, for preparing the hybridization probes The mixed probes were then hybridized to the cDNA microarray After high stringent washing, the cDNA, microarray was scanned for the fluorescent signals and showed the differences between the cell lines Results Among the 8?000 target genes, there were 1?892 (23 65%) genes expressing differently between MCF 7 and LCC2 cell lines Bioinformational analysis on those genes was performed Conclusion DNA microarray technology is an effective technique in screening for differentially expressed genes between different tissues and cell lines
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Objective To study function and screen differentially expressed genes in development of human glioblastoma by using cDNA microarray.Methods BioStarH140S microarray (including 8 347 old genes and 5 592 novel genes) was adopted and hybridized with probes which were prepared for total RNA. These probes were isolated from 2 cases of normal adult brain tissues and 18 cases of glioma tissues. Differentially expressed genes between normal tissues and glioma tissues were analyzed after scanning microarray with ScanArray4 000. These genes were studied with Hierarchical method and Northern hybridization was used for the function of genes. Results Among the 13 939 target genes, there were multiple differentially expressed genes and 5 genes were significantly up-regulated in glioblastoma. Two of them were testified by Northern-blot.Conclusion cDNA microarray technology is a powerful technique in screening differentially expressed genes; 5 up-regulated genes would be helpful to understand the molecular mechanism of glioblastoma. These genes were tightly related to invasion of glioblastoma and might become molecular index of prognosis of glioblastoma.
Gene chip analysis
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Objective:To investigate the differentially expressed genes in hepatic cellular carcinoma(HCC)using cDNA microarray and screen for HCC-related genes.Methods:Total RNA was extracted from normal human liver tissues and hepatic cellular carcinoma tissues.The mRNA was used to prepare probes. The mixed probes were hybridized to the cDNA microarray. After washing, the DNA chips were scanned by laser scanner, The acquired was analyzed.Results:Among the 9182 target genes, 102 genes were identified in HCC tissues that were either over or under-expressed as compared to controls.Forty two genes were up-regulated and 60 down-regulated.Twelve new genes were found in the study. Conclusion: The cDNA microarray for analysis of gene expression profile is a powerful method for screening hepatic cellular carcinoma-related genes.
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Objective: To understand the molecular pathophysiology of hepatocellular carcinoma and pancreatic cancer. Methods: We studied novel gene expression by cDNA microarray method. The PCR products of 4 096 genes and 12 800 gene were spotted onto a kind of chemical-material-coated-glass slide in array. Both the mRNAs from 5 cases of hepatocellular carcinoma and 3 cases of pancreatic cancer were reversely transcribed to cDNAs with the incorporation of fluorescent-labeled dUTP to prepare the hybridization probes. After hybridization, BioDoor4096 and BioDoorl2800 cDNA microarray were scanned for the fluorescent intensity. Tumor invasion-related gene expression was screened through the analysis of difference in gene expression profile. Results: Among 4096 and 12 800 target genes, there were 15 genes whose expression level differed from normal and carcinoma tissues. Therefore, they might be associated with metastasis. Conclusion: Further analysis of these differentially expressed metastasis-associated genes will be helpful for understanding the molecular mechanism of malignant carcinoma.
Gene chip analysis
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Objective: To study the gene expression of high metastatic human ovarian carcinoma cell line (HO-8910PM) and to screen for novel metastasis- associated genes by cDNA microarray. Methods: The cDNA was retro-transcribed from equal quantity mRNA derived from tissues of highly metastatic ovarian carcinoma cell line and normal ovarian, and was labeled with Cy5 and Cy3 fluorescence as probes. The mixed probes were hybridized with BioDoor 4096 double dot human whole gene chip. The chip was scanned by scanArray 3000 laser scanner. The acquired image was analyzed by ImaGene 3.0 software. Results: By applying the cDNA microarray we found: A total of 323 genes whose expression level were 3 times higher or lower in HO-8910PM cell than normal ovarian epithelium cell were screened out, with 71 higher and 252 lower respectively. Among these 10 were new genes. 67 genes showed expression difference bigger than 6 times between HO-8910PM cell and normal ovarian epithelium cell, among these genes 12 were higher, 55 lower, and two new genes were found. Conclusion: cDNA microarray technique is effective in screening the differentially expressed genes between human ovarian cancer cell line (HO-8910PM) and normal ovarian epithelium cell. Using the cDNA microarray to analyze of human ovarian cancer cell line gene expression profile difference will help the gene diagnosis, treatment and protection.
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