Objoctive To identify differential genes between normal ovarian epithelium tissue and ovarian epithelial cancer using representational difference analysis of cDNA （cDNA-RDA）. Methods cDNA-RDA was performed to ident...Objoctive To identify differential genes between normal ovarian epithelium tissue and ovarian epithelial cancer using representational difference analysis of cDNA （cDNA-RDA）. Methods cDNA-RDA was performed to identify the differentially expressed sequences between cDNAs from cancer tissue and cDNAs from normal ovarian tissue in the same patient who was in the early stage of ovarian serous cystadenocarcinoma. These differentially expressed fragments were cloned and analyzed, then sequenced and compared with known genes. Results Three differentially cxpressed cDNA fragments were isolated using cDNA from normal ovarian tissue as tester and cDNA from cancer tissue as driver amplicon by cDNA-RDA. DP Ⅲ- 1 and DP Ⅲ-2 cDNA clone showed significant homology to the cDNA of alpha actin gene; DPⅢ-3 cDNA clone showed significant homology to the cDNA oftransgelin gene. Conclusion cDNA-RDA can bc used to sensitively identify the differentially expressed genes in ovarian serous cystadenocarcinoma. Ovarian serous cystadenocarcinoma involves alteration of multiple genes.展开更多
Objective： To identify aberrant gene expression in primary human T-cell acute lymphoblastic leukemia （T-ALL） and to evaluate the differently expressed level of Lyll and LMO2 genes in human LMO2 positive/TALl negati...Objective： To identify aberrant gene expression in primary human T-cell acute lymphoblastic leukemia （T-ALL） and to evaluate the differently expressed level of Lyll and LMO2 genes in human LMO2 positive/TALl negative T-ALL tumors. Methods： Three methods, representational difference analysis （RDA） of cDNA, cDNA microarrays and RT-PCR were used to detect if Lyll and LMO2 genes were differently expressed in human LMO2 positive/tall negative T-ALL tumors. Results： The results of cDNA RDA and cDNA array shown that Lyll and LMO2 genes are differently expressed in human T-cell tumors. The result of RT-PCR also shown Lyll and LMO2 are high expressed in human T-cell tumors and very low level or no expressed in normal group. Conclusion： We have found that cDNA RDA and cDNA microarray can be successfully used to identify aberrant gene expression in T-ALL cells. In the study described in this manuscript we found that Lyll and LMO2 are aberrantly expressed in human T-ALL LMO2 positive/TALl negative T-ALL tumors.展开更多
Objective To search differentially expressed sequences correlated with pathogenesis of human nasopharyngeal carcinoma (NPC), including the candidates of tumor suppressor genes Methods Representational difference a...Objective To search differentially expressed sequences correlated with pathogenesis of human nasopharyngeal carcinoma (NPC), including the candidates of tumor suppressor genes Methods Representational difference analysis (RDA) was performed to isolate differentially expressed sequences between cDNA from normal human primary cultures of nasopharyngeal epithelial cells and cDNA from NPC cell line HNE1 The source of differentially expressed products were proved by Southern blot, Northern blot and in situ hybridization The fragments were cloned with pGEM T easy kit and sequenced by the chain termination reaction Results Four differentially expressed cDNA fragments were isolated in the fourth subtractive hybridization using cDNA from normal human nasopharyngeal epithelial cells as tester amplicon and cDNA from NPC cell line HNE1 as driver amplicon by cDNA RDA These differential cDNA fragments revealed that they really came from the tester amplicon and were not expressed or down regulated in the NPC HNE1 cells Some of the genes were expressed only in human nasopharyngeal epithelial cells but deleted or down regulated in the biopsies of NPC Of these obtained clones, some were the sequences of the human known genes including house keeping genes, the others represented novel gene sequences Conclusion The differentially expressed products including the candidates of tumor suppressor genes may be associated with the initiation of the NPC展开更多
文摘Objoctive To identify differential genes between normal ovarian epithelium tissue and ovarian epithelial cancer using representational difference analysis of cDNA （cDNA-RDA）. Methods cDNA-RDA was performed to identify the differentially expressed sequences between cDNAs from cancer tissue and cDNAs from normal ovarian tissue in the same patient who was in the early stage of ovarian serous cystadenocarcinoma. These differentially expressed fragments were cloned and analyzed, then sequenced and compared with known genes. Results Three differentially cxpressed cDNA fragments were isolated using cDNA from normal ovarian tissue as tester and cDNA from cancer tissue as driver amplicon by cDNA-RDA. DP Ⅲ- 1 and DP Ⅲ-2 cDNA clone showed significant homology to the cDNA of alpha actin gene; DPⅢ-3 cDNA clone showed significant homology to the cDNA oftransgelin gene. Conclusion cDNA-RDA can bc used to sensitively identify the differentially expressed genes in ovarian serous cystadenocarcinoma. Ovarian serous cystadenocarcinoma involves alteration of multiple genes.
文摘Objective： To identify aberrant gene expression in primary human T-cell acute lymphoblastic leukemia （T-ALL） and to evaluate the differently expressed level of Lyll and LMO2 genes in human LMO2 positive/TALl negative T-ALL tumors. Methods： Three methods, representational difference analysis （RDA） of cDNA, cDNA microarrays and RT-PCR were used to detect if Lyll and LMO2 genes were differently expressed in human LMO2 positive/tall negative T-ALL tumors. Results： The results of cDNA RDA and cDNA array shown that Lyll and LMO2 genes are differently expressed in human T-cell tumors. The result of RT-PCR also shown Lyll and LMO2 are high expressed in human T-cell tumors and very low level or no expressed in normal group. Conclusion： We have found that cDNA RDA and cDNA microarray can be successfully used to identify aberrant gene expression in T-ALL cells. In the study described in this manuscript we found that Lyll and LMO2 are aberrantly expressed in human T-ALL LMO2 positive/TALl negative T-ALL tumors.
文摘Objective To search differentially expressed sequences correlated with pathogenesis of human nasopharyngeal carcinoma (NPC), including the candidates of tumor suppressor genes Methods Representational difference analysis (RDA) was performed to isolate differentially expressed sequences between cDNA from normal human primary cultures of nasopharyngeal epithelial cells and cDNA from NPC cell line HNE1 The source of differentially expressed products were proved by Southern blot, Northern blot and in situ hybridization The fragments were cloned with pGEM T easy kit and sequenced by the chain termination reaction Results Four differentially expressed cDNA fragments were isolated in the fourth subtractive hybridization using cDNA from normal human nasopharyngeal epithelial cells as tester amplicon and cDNA from NPC cell line HNE1 as driver amplicon by cDNA RDA These differential cDNA fragments revealed that they really came from the tester amplicon and were not expressed or down regulated in the NPC HNE1 cells Some of the genes were expressed only in human nasopharyngeal epithelial cells but deleted or down regulated in the biopsies of NPC Of these obtained clones, some were the sequences of the human known genes including house keeping genes, the others represented novel gene sequences Conclusion The differentially expressed products including the candidates of tumor suppressor genes may be associated with the initiation of the NPC
