EFFECT OF TELOMERASE ANTISENSE hTERT ON TELOMERASE ACTIVITY AND PROLIFERATION OF K562 CELLS

Objective:To construct a plasmid,antisense hTERT plasmid then transferred it into K562 cells,and to see if it can inhibit the activity of telomerase and the proliferation of K562 cells.Methods:Plasmid construction and transfection.An antisense plasmid was constructed by reverse design of PCR primers was cloned to the plasmid pLNCX neo,G418 was added into the medium after the plamid was successfully introduced into K562 cells by using lipofectactin mediated DNA transfection.Analysis the effects of antisense hTERT on K562 cells.The effects of telomerase inhibition on the proliferation of K562 cells were analyzed by means of MTT and morphology.The telomerase activity of K562 cells was studied by TRAP PCR ELISA methods.Results:The growth rate of antisense hTERT gene transfected K562 cells (K562 as cell) was significantly slower than those of the controls,and apoptosis appear in part of K562 as cells the telomerase activity of antisense hTERT gene transfected cells was significantly inhibited than those of controls.Conclusion:The expression of a partial compliment antisense sequence to the mRNA sequence of the protein subunit of telomerase can inhibit the activity of telomerase,slow the growth of K562 cells.