AIM:To investigate the effect of transfected survivin antisense oligonucleotide (ASODN) on proliferation and apoptosis of gastric cancer ceils. METHODS:The authors designed ASODNs targeting different regions of survMn...AIM:To investigate the effect of transfected survivin antisense oligonucleotide (ASODN) on proliferation and apoptosis of gastric cancer ceils. METHODS:The authors designed ASODNs targeting different regions of survMn mRNA,including surviving ASODN1,ASODN2 and ASODN3.ASODNs were transfected into gastric cancer cell line SGC 7901,cell growth was detected by MTT assay.Cells exposed to the potent oligonucleotide were also examined for apoptosis induction by FCM and fluorescence microscopy.Semiquantitive RT-PCR and Western blot examinations were carried for expression of survivin mRNA and protein. RESULTS:ASODN3 caused a statistically significant reduction of cell viability to 60.6% (±2.9%) (P<0.01),while ASODN1 and ASODN2 had no such changes (P>0.05).The cell growth was also significantly inhibited by ASODN3, compared with reversal and scrambled sequence.A significant loss of survivin mRNA was presented in ASODN3 treated ceils and this was not seen in treatment with sense ODN or scramble ODN.Protein level was significantly decreased 48 h after survivin ASODN trasfected by approximately 2-fold decrease comparedwith untreated controls.However. ASODN3 did not induce significant apoptosis response until 48 h after transfection (P>0.05). CONCLUSION:ASODN3,which targets translation initiation part,can be identified as a most potent antisense compound. Srvivin ASODN3 may provide a novel approach to therapy of gastric cancer.展开更多
AIM:To evaluate the inhibitory effect of antisense phosphorothioate oligonucleotide (asON) complementary to the initiator of human telomerase catalytic subunit (hTERT) on the growth of hepatoma cells.METHODS:The as-hT...AIM:To evaluate the inhibitory effect of antisense phosphorothioate oligonucleotide (asON) complementary to the 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 ashTERT at the concentration of 10μmol/L. After 72h, these cells were obtained for detecting growth inhibition,telomerase activity using the methods of MTT,TRAP-PCR-ELISA, respectively. BALB/c(nu/nu) mice were injected HepG2.2.15 cells and a human-nude mice model was obtained. There were three groups for anti-tumor activity study. Once tumors were established, these animals in the first group were administered as-hTERT and saline.Apoptosis of tumor cells was detected by FCM. In the 2nd group, the animals were injected HepG2.2.15 cells together with as-hTERT. In the third group, the animals were given as-hTERT 24 hours postinjection of HepG2.2.15 cells. The anti-HBV effects were assayed with ELISA in vitro and in vivo.RESULTS: Growth inhibition was observed in cells treated with as-hTERT in vitro. A significant different in the value of A570-A630 was found between cells treated with as-hTERT and control (P<0.01) by MTT method. The telomerase activity of tumor cells treated with as-hTERT was reduced,the value of A4so nm was 0.42 compared to control (1.49) with TRAP-PCR-ELISA. The peak of apoptosis in tumor cells given as-hTERT was 21.12%, but not seen in saline-treated control. A prolonged period of carcinogenesis was observed in the second and third group animals. There was inhibitory effect on the expression of HBsAg and HBeAg in vivo and in vitro.CONCLUSION: As-hTERT has an anti-tumor activity, which may be useful for gene therapy of tumors.展开更多
基金Supported by Shanghai Key Research Project Grant in Medical Science Development (99ZD003)
文摘AIM:To investigate the effect of transfected survivin antisense oligonucleotide (ASODN) on proliferation and apoptosis of gastric cancer ceils. METHODS:The authors designed ASODNs targeting different regions of survMn mRNA,including surviving ASODN1,ASODN2 and ASODN3.ASODNs were transfected into gastric cancer cell line SGC 7901,cell growth was detected by MTT assay.Cells exposed to the potent oligonucleotide were also examined for apoptosis induction by FCM and fluorescence microscopy.Semiquantitive RT-PCR and Western blot examinations were carried for expression of survivin mRNA and protein. RESULTS:ASODN3 caused a statistically significant reduction of cell viability to 60.6% (±2.9%) (P<0.01),while ASODN1 and ASODN2 had no such changes (P>0.05).The cell growth was also significantly inhibited by ASODN3, compared with reversal and scrambled sequence.A significant loss of survivin mRNA was presented in ASODN3 treated ceils and this was not seen in treatment with sense ODN or scramble ODN.Protein level was significantly decreased 48 h after survivin ASODN trasfected by approximately 2-fold decrease comparedwith untreated controls.However. ASODN3 did not induce significant apoptosis response until 48 h after transfection (P>0.05). CONCLUSION:ASODN3,which targets translation initiation part,can be identified as a most potent antisense compound. Srvivin ASODN3 may provide a novel approach to therapy of gastric cancer.
基金Supported by the National Natural Science Foundation of China,No.30070341
文摘AIM:To evaluate the inhibitory effect of antisense phosphorothioate oligonucleotide (asON) complementary to the 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 ashTERT at the concentration of 10μmol/L. After 72h, these cells were obtained for detecting growth inhibition,telomerase activity using the methods of MTT,TRAP-PCR-ELISA, respectively. BALB/c(nu/nu) mice were injected HepG2.2.15 cells and a human-nude mice model was obtained. There were three groups for anti-tumor activity study. Once tumors were established, these animals in the first group were administered as-hTERT and saline.Apoptosis of tumor cells was detected by FCM. In the 2nd group, the animals were injected HepG2.2.15 cells together with as-hTERT. In the third group, the animals were given as-hTERT 24 hours postinjection of HepG2.2.15 cells. The anti-HBV effects were assayed with ELISA in vitro and in vivo.RESULTS: Growth inhibition was observed in cells treated with as-hTERT in vitro. A significant different in the value of A570-A630 was found between cells treated with as-hTERT and control (P<0.01) by MTT method. The telomerase activity of tumor cells treated with as-hTERT was reduced,the value of A4so nm was 0.42 compared to control (1.49) with TRAP-PCR-ELISA. The peak of apoptosis in tumor cells given as-hTERT was 21.12%, but not seen in saline-treated control. A prolonged period of carcinogenesis was observed in the second and third group animals. There was inhibitory effect on the expression of HBsAg and HBeAg in vivo and in vitro.CONCLUSION: As-hTERT has an anti-tumor activity, which may be useful for gene therapy of tumors.
