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AIM:To evaluate the inhibitory effect of antisensephosphorothioate oligonucleotide(asON)complementary tothe initiator of human telomerase catalytic subunit(hTERT)on the growth of hepatoma cells.METHODS:The as-hTERT was synthesized by using aDNA synthesizer.HepG2.2.15 cells were treated with as-hTERT at the concentration of 10μmol/L.After 72h,thesecells were obtained for detecting growth inhibition,telomerase activity using the methods of MTF,TRAP-PCR-ELISA,respectively.BALB/c(nu/nu)mice were injectedHepG2.2.15 cells and a human-nude mice model wasobtained.There were three groups for anti-tumor activitystudy.Once tumors were established,these animals inthe first group were administered as-hTERT and saline.Apoptosis of tumor cells was detected by FCM.In the2nd group,the animals were injected HepG2.2.15 cellstogether with as-hTERT.In the third group,the animalswere given as-hTERT 24 hours postinjection of HepG2.2.15cells.The anti-HBV effects were assayed with ELISA in vitroand in vivo.RESULTS:Growth inhibition was observed in cells treatedwith as-hTERT in vitro.A significant different in the valueof A_(570)-A_(630) was found between cells treated with as-hTERTand control(P<0.01)by MTT method.The telomeraseactivity of tumor cells treated with as-hTERT was reduced,the value of A_(450) nm was 0.42 compared to control(1.49)with TRAP-PCR-ELISA.The peak of apoptosis in tumor cellsgiven as-hTERT was 21.12%,but not seen in saline-treatedcontrol.A prolonged period of carcinogenesis was observedin the second and third group animals.There was inhibitoryeffect on the expression of HBsAg and HBeAg in vivo andin vitro.CONCLUSION:As-hTERT has an anti-tumor activity,whichmay be useful for gene therapy of tumors.
AIM: To evaluate the inhibitory effect of antisense phosphorothioate oligonucleotide (asON) complementary tothe initiator of human telomerase catalytic subunit (hTERT) on the growth of hepatoma cells. METHODS: The as-hTERT was synthesized by using a DNA synthesizer. HepG2.2.15 cells were treated with as-hTERT at the concentration of 10 μmol / L.After 72 h, these cells were obtained for detecting growth inhibition, telomerase activity using the methods of MTF, TRAP-PCR-ELISA, respectively.BALB / c (nu / nu) mice were injectedHepG2 .2.15 cells and a human-nude mice model wasobtained.There were three groups for anti-tumor activity study. Oncions were established, these animals inthe first group were administered as-hTERT and saline. Apoptosis of tumor cells was detected by FCM. the2nd group, the animals were injected HepG2.2.15cellstogetherwithas-hTERT.In the third group, the animalswere given as-hTERT 24 hours postinjection of HepG2.2.15cells. The anti-HBV effects were assayed with ELISA in vitro and in vivo. RE SULTS: Growth inhibition was observed in cells treated with as-hTERT in vitro. A significant different in the value of A_ (570) -A_ (630) was found between cells treated with as-hTERTand control (P <0.01) telomerase activity of tumor cells treated with as-hTERT was reduced, the value of A_ (450) nm was 0.42 compared to control (1.49) with TRAP-PCR-ELISA.The peak of apoptosis in tumor cells as live-hTERT was 21.12% not seen in saline-treatedcontrol. A prolonged period of carcinogenesis was observed in the second and third group animals. Here was inhibitory effect on the expression of HBsAg and HBeAg in vivo and in vitro. CONCLUSION: As-hTERT has an anti-tumor activity, whichmay be useful for gene therapy of tumors.