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AIM:To investigate the inhibitory effect of phosphorothioateanti-sense oligodeoxynucleotides(PASODN)on colorectalcancer LS-174T ceils in vitro and the mechanism of inhibitionof telomerase activity in these cells.METHODS:PASODN were used to infect LS-174T cellsand block human telomerase RNA(hTR)through anti-sensetechnology.The inhibitory effect of PASODN was evaluatedby colony-forming inhibition assay and growth curve.Changesof telomerase activity in LS-174T cells were detected bypolymerase chain reaction-enzyme-linked immunosorbentassay(PCR-ELISA),and the level of apoptosis was analyzedby flow cytometry(FCM)assay.RESULTS:PASODN showed a dose and time-dependentinhibition of cell proliferation.The optimal dosage of PASODNwas 10 μmol/L.The colony-forming efficiency was 10.3% inPASODN group after 10 d,whereas that in phosphorothioatemis-sense oligodeoxynudeotides(PMSODN)group with thesame concentration and in PBS group(blank control)was49.1% and 50.7%,respectively,PCR-ELISA results indicatedthat telomerase activity in the PASODN group was obviouslyinhibited in comparison with in the control groups(P<0.02,t=3.317 and 3.242,t_(0.01)(20)=2.845).Meanwhile,beforethe number of cells was decreased,the morphologicalchanges were observed in the cells of PASODN group.Thecells in PASODN group showed the apoptotic peak at 72 hafter infection,whereas the control group did not show.CONCLUSION:Specific sequence oligonucleotides caninhibit telomerase activity and lead to cell apoptosis,suggesting a novel treatment strategy for malignant tumorsinduced by telomerase.
AIM: To investigate the inhibitory effect of phosphorothioate-sense oligodeoxynucleotides (PASODN) on colorectal cancer LS-174T ceils in vitro and the mechanism of inhibition of telomerase activity in these cells. METHODS: PASODN were used to infect LS- 174T cells and block human telomerase RNA hTR) through anti-sense technology. The inhibitory effect of PASODN was evaluated by colony-forming inhibition assay and growth curve. Changes in telomerase activity in LS-174T cells were detected by polymerase chain reaction-enzyme-linked immunosorbent assay (PCR-ELISA) of PASODN was assayed by flow cytometry (FCM) assay .RESULTS: PASODN showed a dose and time-dependent inhibition of cell proliferation. The optimal dosage of PASODNwas 10 μmol / L.The colony-forming efficiency was 10.3% inPASODN group after 10 d, that in phosphorothioatemis-sense oligodeoxynucleotides (PMSODN) group with the concentration and in PBS group (blank control) was 49.1% and 50.7% respectively, PCR-ELISA results indica tedthat telomerase activity in the PASODN group was obviously inhibited in comparison with the control groups (P <0.02, t = 3.317 and 3.242, t 0.01 (20) = 2.845), while the number of cells was decreased, the morphological changes were observed in the cells of PASODN group. The cells in PASODN group. The cells in PASODN group showed the apoptotic peak at 72 hafter infection, whereas the control group did not show. CONCLUSION: Specific sequence oligonucleotides canhibit telomerase activity and lead to cell apoptosis, suggesting a novel treatment strategy for malignant tumorsinduced by telomerase.