Acknowledgments
The authors would like to thank everyone in Biology Department and Chemistry Department of CytRx Corporation for their strong supports during the project.
Keywords
Telomerase; hTERT; VEGF
Introduction
Human telomerase is a ribonucleoprotein complex composed of the catalytic subunit (hTERT) which adds telomeric repeats onto the ends of chromosomes and a RNA component (hTR) serving as a template for telomeric DNA synthesis [1]. Most human somatic DZNeP have undetectable telomerase activity due to transcriptional repression of the hTERT gene during early embryonic development, but telomerase is reactivated during tumorigenesis and is present in over 90% of tumors [2]. Telomerase is almost universally required for cellular immortality and is permissive for tumorigenesis [3], [4], [5] and [6].
Materials and methods
Cell lines and culture conditions. The human lung fibroblast WI-38 and HeLa cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (Hyclone, Logan, USA), 100 units/ml penicillin, 100 μg/ml streptomycin, and glutamine (Gibco, NY, USA) in 5% CO2 at 37 oC. WI-38/GFP and WI-38/hTERT stable cell lines were established by infection of WI-38 cells with retroviruses expressing EGFP or hTERT, and selected by 100 μg/ml hygromycin B (Roche, Basel, Switzerland).
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