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AIM:To study the effects of doxorubicin on telomeraseactivity and telomere length in hepatocellular carcinoma.METHODS:Telomerase activity was assayed with a non-radioisotopic quantitative telomerase repeat amplificationprotocal-based method.The effect of doxorubicin(DOX)onthe growth of BEL-7404 human hepatoma cells wasdetermined by microculture tetrazolium assay.Meantelomere length(terminal restriction fragment)was detectedby Southern blot method.The expression of telomerasesubunits genes was investigated by RT-PCR.Cell apoptosisand cell cycle distribution were evaluated by flow cytometry.RESULTS:Telomerase activity was inhibited in a dose andtime-dependent manner in BEL-7404 human hepatoma cellstreated with DOX for 24,48 or 72 h in concentrations from0.156 to 2.5 μM which was crrelated with the inhibition ofcell growth.No changes were found in the mRNA expressionof three telomerase subunits(hTERT,hTR and TP1)afterdrug exposure for 72 h with indicated concentrations.Thecells treated with DOX showed shortened mean telomerelength and accumulated at the G_2/M phase.However,therewas almost no effects on cell apoptosis by DOX.CONCLUSION:The telomerase inhibition and the telomereshortening by DOX may contribute to its efficiency in thetreatment in hepatocellular carcinoma.
The effect of doxorubicin on the growth of BEL-7404 human Hepatoma cells wasdetermined by microculture tetrazolium assay.Meantelomere length (terminal restriction fragment) was detected by Southern blot method.The expression of telomerasesubunits genes was investigated by RT-PCR.Cell apoptosis and cell cycle distribution was evaluated by flow cytometry.RESULTS:Telomerase activity was inhibited In a dose andtime-dependent manner in BEL-7404 human hepatoma cells treated with DOX for 24,48 or 72 h in concentrations from0.156 to 2.5 μM which was cr associated with the inhibition ofcell growth.No changes were found in the mRNA expressionof three telomerase Subunits(hTERT,hTR and TP1)afterdrug exposure for 72 h with indicated concentrations.The cellss treated w Ith DOX shorten shortened mean telomerelength and accumulated at the G_2/M phase.However, therewas almost no effects on cell apoptosis by DOX.CONCLUSION:The telomerase inhibition and the telomereshortening by DOX may contribute to its efficiency in thetreatment in hepatocellular carcinoma.