Inhibition of Cell Growth and Telomerase Activity in Osteosarcoma Cells by DN-hTERT
- Author:
Tao XU
1
;
Yaojian RAO
;
Wentao ZHU
;
Fengjin GUO
Author Information
1. 华中科技大学同济医学院附属同济医院
- Keywords:
dominant negative human telomerase reverse transcriptase;
MG63;
telomerase activity
- From:
Journal of Huazhong University of Science and Technology (Medical Sciences)
2006;26(5):601-603
- CountryChina
- Language:Chinese
-
Abstract:
In order to study the effects of dominant negative human telomerase reverse transcriptase (DN-hTERT) on cell growth and telomerase activity in osteosarcoma cell line MG63, MG63 cells were transfected with DN-hTERT-IRES2-EGFP9 (DN) or IRES2-EGF (I, blank vector) with lipofectamine 2000. The stably transfected cells were selected with G-418. Cell growth properties were examined under a fluorescence microscope. The hTERT mRNA expression was detected by reverse transcription-polymerase chain reaction (RT-PCR). Telomerase activities were measured by TRAP-ELISE. The tumorigenicity was studied with tumor xenografts by subcutaneous injection of cancer cells into nude mice. The results showed that cell growth was suppressed in MG63 cells transfected with DN-hTERT. The hTERT mRNA was increased in N-hTERT transfected-MG63 cells (MG63/DN). The telomerase activity was 2.45±0.11 in MG63/DN cells, while 3.40±0.12 in the cells transfected with blank vector (MG63/I), (P＜0.05); DN-hTERT-expressing clones did not form tumors in 2 weeks, but the ratio of tumorigenesis was 30 % in nude mice bearing MG63/I (P＜0.01). It was concluded that DN-hTERT could specifically inhibit the cell growth and telomerase activity in MG63 cells.
