Inhibition of telomerase activity on breast cancer cells by dominant negative human telomerase reverase transcriptase gene / 医学研究生学报

ABSTRACT

Objective:To explore inhibition of telomerase activity and malignant phenotype on breast cancer cells MCF7 by Dominant Negative human telomerase reverase transcriptase gene(DN-hTERT).Methods:DN-hTERT eukaryotic expression vector DN-hTERT-IRES2-EGFP and empty vector(I) IRES2-EGFP were transfected into MCF7 by lipofectamine2000,after being selected by G418,positive clones were obtained;Tthe transfected cells growth was observed with inverted fluorescence microscope.The expression levels of hTERT mRNA of transfected cells were determined by reverse transcriptase polymerase chain reaction(RT-PCR).Telomerase activity of transfected cells was measured by TRAP-ELISE.Results:The proliferation of MCF7/DN cells were inhibited by DN-hTERT transtection;The expression levels of hTERT mRNA increased in MCF7/DN.Telomeric length was shorter in MCF7/DN than that in MCF7/I.The telomerase activity measured by TRAP-ELISE was 2.36?0.12 in MCF7/DN and was 3.32?0.14 in vacant vector MCF7/I.The telomerase activity in MCF7/DN was significantly lower than in vacant vector transfected MCF7/I(P