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Detection of telomerase activity in human cells with telomeric repeat amplification protocol by silver staining 被引量:1

Detection of telomerase activity in human cells with telomeric repeat amplification protocol by silver staining
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摘要 Telomerase is a ribonucleoprotein enzyme, which synthesizes telomeric repeats (TTAGGG) n.While germline cells and most malignant tumor cells express telomerase activity, normal somatic cells aregenerally deficient in telomerase activity. Our objective was to detect telomerase activity of human cells bysilver staining with telorneric repeat amplification protocol (TRAP) which is easy and quick. Comparing withradioisotopic TRAP, we examined the telomerase activity in telomerase-positive 293-cell and RNase-pretreated and heat-pretreated negative controls by silver staining TRAP. We detected telomerase activity in 2 strainsof human liver tumor cells (QGY7701 and SMMC7721 ). The 293 cells (only 10 cells) and the 2 strains ofhuman liver tumor cells were all positive. while telomerase activity was not detected in the negative controls.These data suggest that non-radioisotopic silver staining TRAP is a specific, sensitive and fast assay fortelomerase activity. It was verified that the 2 strains of human liver tumor cells express telomerase activity. Telomerase is a ribonucleoprotein enzyme, which synthesizes telomeric repeats (TTAGGG) n.While germline cells and most malignant tumor cells express telomerase activity, normal somatic cells aregenerally deficient in telomerase activity. Our objective was to detect telomerase activity of human cells bysilver staining with telorneric repeat amplification protocol (TRAP) which is easy and quick. Comparing withradioisotopic TRAP, we examined the telomerase activity in telomerase-positive 293-cell and RNase-pretreated and heat-pretreated negative controls by silver staining TRAP. We detected telomerase activity in 2 strainsof human liver tumor cells (QGY7701 and SMMC7721 ). The 293 cells (only 10 cells) and the 2 strains ofhuman liver tumor cells were all positive. while telomerase activity was not detected in the negative controls.These data suggest that non-radioisotopic silver staining TRAP is a specific, sensitive and fast assay fortelomerase activity. It was verified that the 2 strains of human liver tumor cells express telomerase activity.
出处 《Journal of Medical Colleges of PLA(China)》 CAS 1998年第2期140-142,共3页 中国人民解放军军医大学学报(英文版)
关键词 TELOMERE TELOMERASE SILVER STAINING telomere telomerase silver staining
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  • 1Mieczyslaw A. Piatyszek,Nam W. Kim,Scott L. Weinrich,Keiko Hiyama,Eiso Hiyama,Woodring E. Wright,Jerry W. Shay PhD.Detection of telomerase activity in human cells and tumors by a telomeric repeat amplification protocol (TRAP)[J]. Methods in Cell Science . 1995 (1)
  • 2Iakian VA.Telomerase: Beginning to understand the end. Science . 1995
  • 3Hiyama E,Yokoyama T,Tatsumoto N et al.Telomerase activityin gastric cancer.Cant. er Res . 1995
  • 4Morin GB.The human telomere terminal transferase enzyme is aribonucleoprotein that synthesizes TTAGGG repeats. Cell . 1989
  • 5Kim NW,Piatyszek MA,Prowse KR at al.Specific associationof human telomerase activity with immortal cells and cancer. Science . 1994
  • 6Chadeneau C,Hay K,Hirte HW et al.Telomerase activity associated with acquisition of malignancy in human colorectal cancer. Cancer Research . 1995
  • 7Tahara H,Nakanishi T,Kitamoto M et al.Telomerase activityin human liver tissues: Comparison between chronic liver diseaseand hepatocellular carcinomas. Cancer Research . 1995
  • 8Hiyama K,Hiyama E,Yokoyama T et al.Correlating telomeraseactivity levels with human neuroblastoma outcomes. Nature Medicine . 1995

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