Effects of mutation in Cosmc non-coding region on the transcription level of Cosmc mRNA in Tn antigen positive tumor cells / 中华微生物学和免疫学杂志

Objective To explore the mutation in corel β3-galactosyl-transferase specific molecular chaperone(Cosmc、no-coding region and it's effects on the transcription level of Cosmc in Tn antigen positive tumor cells.Methods The Tn antigen positive(Tn+)and negative(Tn-)cells were separated from tumor tissues by immune magnetic bead,then the genomic DNA(gDNA),total RNA were prepared by Qiagen AllPrep DNA/RNA mini kit. In these cells.the transcription levels of T-synthase and Cosmc mRNA were tested by RT-PCR.the DNA of Cosmc non-coding region was amplified by PCR,the mutation in Cosmc non-coding region were further detected by sequencing.Results There are no mutation appearing in Tn-cells,one or more mosaic sequence allele appearing in portion of patient's Tn-cells.Almost of the Tn+cells which separated from tumor tissues and Jurkat T cell exists mutation.but the mutation style and mutation point were not saine in different tumor.Thtee patient's Tn+cells have loss of hetemzygosity(LOH),four patient's Tn+cells and Jurkat T cell have point mutation.Although no difference of transcription level of T-synthase mRNA in Tn+ and Tn-cells.but the transcription level of Cosmc mRNA in Tn+ cell was much lower than that in Tn-cell.The ratio of T-synthase/Cosmc mRNA in Tn+ tumor cells was hiigher than that in Tn-cell.Conclusion The tumor Tn antigen arise from mutation in Cosmc non-coding region maybe result from transcription level decreased of Cosmc mRNA.

Loss of heterozygosity of microsatellite DNA on 6q in bladder tumor / 中国康复理论与实践

@#ObjectiveTo investigate the loss of heterozygosity (LOH) on 6q in bladder tumor.MethodsD6S404 and D6434 microsatellite markers near 6q21 were tested by PCR-SSLP-stain method on tumor DNA from 31 cases of bladder tumor.ResultsAmong these 31 cases of bladder tumor,LOH was detected in tumor tissues on site for D6S404 (35.5%) and D6S434(22.6%).ConclusionOne or more tumor suppressor gene near 6q21 maybe relevant for the development of bladder tumor.

Loss of Heterozygosity in Endometriosis / 대한산부인과학회잡지

OBJECTIVE: Endometriosis is a very common gynecological condition in which tissue similar to endometrium proliferates at sites outside the uterine cavity. Although it generally remain a benign condition, malignant transformation has been documented, and it is commonly found in association with endometrioid subtype ovarian carcinoma. In order to identify the genomic change in those areas possibly involved in the pathogenesis of endometriosis, we performed LOH analysis. METHODS: Twenty seven cases of endometriosis were analyzed for the detection of LOH using 5 microsatellite markers. LOH analysis was performed by PCR, capillary electrophoresis and gene scan analysis using DNA from sections of tumor and normal tissue pairs. RESULTS: Twenty two of 27 (81.5%) cases demonstrated LOH at one or more loci. The frequency of LOH was 37.0% (D18S69), 25.9% (D22S274), 14.8% (D22S283), 7.4% (D6S286), 7.4% (D13S160). CONCLUSION: The frequencies of LOH was increased in higher stage of endometriosis. Most notable findings were found at chromosome 18 and 22 loci (D18S69, D22S274). These region might involve the some candidate genes closely related with the pathogenesis of endometriosis.

Loss of Heterozygosity on Chromosome 10, 13, 17 and p53 Gene Mutations in Human Brain Gliomas

Gliomas, the most common primary tumors of the human central nervous system, are usually malignant and virtually incurable. They can be classified according to their cellular differentiation:astrocytoma, oligodendroglioma, and ependymoma. The majority of these brain tumors are astrocytomas, which typically progress through three histopathologically defined stages with the passage of time:one premalignant stage, low-grade astrocytoma, and two malignant stages, anaplastic astrocytoma and glioblastoma multiforme. Recent studies on the molecular mechanisms of carcinogensis have demonstrated a possible role for two classes of genes in neoplastic transformation:tumor suppressor genes and oncogenes. Tumor suppressor genes are wild-type alleles of genes that are believed to function normally in the cell to suppress cellular proliferation. Inactivation of both copies of suppressor gene may contribute to neoplastic transformation by removing a normal constraint to cell growth. The well characterised suppressor genes are RB gene and p53 gene. Gliomas, like most other cancers, are associated with several genetic changes, including oncogenes and suppressor genes. In an attempt to further our knowledge of tumor suppressor genes contributing glioma development and progression, restriction fragment length polymorphism(RFLP) analysis was done to determine loss of heterozygosity(LOH) on chromosome 10. 13q(RBI), 17p, and 22q containing putative tumor suppressor genes in 36 cases of human gliomas with various malignancy grades. And to detect p53 gene mutations at exon 5, 6, and 7 in 23 cases of malignant gliomas, polymerase chain reaction-single strand conformation polymorphisms(PCR-SSCP) analysis was performed. Loss of heterozygosity for loci on chromosome 10 were found in four of 5(60%) informative cases of glioblastoma multiforme and one of 2(50%) cases of anaplastic astrocytomas. Loss of heterozygosity on chromosome 17p was found in eight of 17(47%) informative cases of malignant gliomas, including 2 cases of anaplastic oligodendroglioma. There was no allelic loss of chromosome 10 and 17 in benign gliomas. Deletions on RBI locus were seen in six of 10(60%) informative cases of glioblastoma multiforme and two of 5(40%) informative cases of low-grade astrocytomas, suggesting that RBI gene may have a role associated with the early events in tumorigenesis. In PCR-SSCP analysis, six of 23(26%) cases of malignant gliomas, including one case of anaplastic oligodendroglioma, showed mobility shifts on exon 5 or 7 of p53 gene which suggest point mutations of this gene. There was no LOH at IGLC2 locus on chromosome 22. On the basis of the data presented here, it is possible to associate certain molecular abnormalities with gliomas of increasing grades of malignancy, deletion of RB gene, loss of heterozygosity on chromosome 17p, p53 gene mutation, and loss of allele on chromosome 10.

Loss of Heterozygosity on Chromosome 10, 13, 17 and p53 Gene Mutations in Human Brain Gliomas

Gliomas, the most common primary tumors of the human central nervous system, are usually malignant and virtually incurable. They can be classified according to their cellular differentiation:astrocytoma, oligodendroglioma, and ependymoma. The majority of these brain tumors are astrocytomas, which typically progress through three histopathologically defined stages with the passage of time:one premalignant stage, low-grade astrocytoma, and two malignant stages, anaplastic astrocytoma and glioblastoma multiforme. Recent studies on the molecular mechanisms of carcinogensis have demonstrated a possible role for two classes of genes in neoplastic transformation:tumor suppressor genes and oncogenes. Tumor suppressor genes are wild-type alleles of genes that are believed to function normally in the cell to suppress cellular proliferation. Inactivation of both copies of suppressor gene may contribute to neoplastic transformation by removing a normal constraint to cell growth. The well characterised suppressor genes are RB gene and p53 gene. Gliomas, like most other cancers, are associated with several genetic changes, including oncogenes and suppressor genes. In an attempt to further our knowledge of tumor suppressor genes contributing glioma development and progression, restriction fragment length polymorphism(RFLP) analysis was done to determine loss of heterozygosity(LOH) on chromosome 10. 13q(RBI), 17p, and 22q containing putative tumor suppressor genes in 36 cases of human gliomas with various malignancy grades. And to detect p53 gene mutations at exon 5, 6, and 7 in 23 cases of malignant gliomas, polymerase chain reaction-single strand conformation polymorphisms(PCR-SSCP) analysis was performed. Loss of heterozygosity for loci on chromosome 10 were found in four of 5(60%) informative cases of glioblastoma multiforme and one of 2(50%) cases of anaplastic astrocytomas. Loss of heterozygosity on chromosome 17p was found in eight of 17(47%) informative cases of malignant gliomas, including 2 cases of anaplastic oligodendroglioma. There was no allelic loss of chromosome 10 and 17 in benign gliomas. Deletions on RBI locus were seen in six of 10(60%) informative cases of glioblastoma multiforme and two of 5(40%) informative cases of low-grade astrocytomas, suggesting that RBI gene may have a role associated with the early events in tumorigenesis. In PCR-SSCP analysis, six of 23(26%) cases of malignant gliomas, including one case of anaplastic oligodendroglioma, showed mobility shifts on exon 5 or 7 of p53 gene which suggest point mutations of this gene. There was no LOH at IGLC2 locus on chromosome 22. On the basis of the data presented here, it is possible to associate certain molecular abnormalities with gliomas of increasing grades of malignancy, deletion of RB gene, loss of heterozygosity on chromosome 17p, p53 gene mutation, and loss of allele on chromosome 10.