[Qualitative and quantitative analyses of telomerase activity in cultured cells].
In this study, we established a qualitative PCR-TRAP assay by using SYBR Green stain instead of EB stain; this modification based on Kim's method raised the sensitivity of telomerase detection by 25-100 fold. Besides, a quantitative assay was established by us using 32P-ATP labeled TS primer and the internal control TSK1. We detected the telomerase activity of 12 cultured cells by means of these two assays. The results showed that the telomerase activity could be detected in all the cells, but the activity levels of the cells differed prominently (from 23 to 652 TPG units). The establishment of PCR-TRAP assay and especially the establishment of quantitative assay have enabled us to evaluate the telomerase activity of cells more accurately, and they can be used in our further studies of tumor gene therapy by reducing the telomerase activity of tumor cells.
Ouyang
X
X
Medical Molecular Biology Laboratory, School of Basic Medical Sciences, Chengdu.
Qu
Y
Y
Liu
S
S
Liu
B
B
D001943:Breast Neoplasms / Q000201:enzymology / Q000473:pathology; D005260:Female; D006367:HeLa Cells / Q000201:enzymology; D006801:Humans; D010051:Ovarian Neoplasms / Q000201:enzymology / Q000473:pathology; D016133:Polymerase Chain Reaction / Q000379:methods; D013274:Stomach Neoplasms / Q000201:enzymology / Q000473:pathology; D019098:Telomerase / Q000032:analysis* / Q000378:metabolism; D014407:Tumor Cells, Cultured / Q000201:enzymology*
D019098:Telomerase