AIM: To evaluate antihepatoma effect of antisense phosphorothioate oligodeoxyribonucleotides (S-ODNs) targeted to alpha-fetoprotein (AFP) genes in vitro and in nude mice. METHODS: AFP gene expression was examined by i...AIM: To evaluate antihepatoma effect of antisense phosphorothioate oligodeoxyribonucleotides (S-ODNs) targeted to alpha-fetoprotein (AFP) genes in vitro and in nude mice. METHODS: AFP gene expression was examined by immunocytochemical method or enzyme-linked immunosorbent assay. Effect of S-ODNs on SMMC-7721 human hepatoma cell growth in vitro was determined using microculture tetrazolium assay. In vitro antitumor activities of S-ODNs were monitored by measuring tumor weight differences in treated and control mice bearing SMMC-7721 xenografts. Induction of cell apoptosis was evaluated by fluorescence-activated cell sorter (FACS) analysis. RESULTS: Antisense S-ODN treatment led to reduced AFP gene expression. Specific antisense S-ODNs, but not control S-ODNs, inhibited the growth of hepatoma cells in vitro. In vitro, only antisense S-ODNs exhibited obvious antitumor activities. FACS analysis revealed that the growth inhibition by antisense S-ODNs was associated with their cell apoptosis induction. CONCLUSION: Antisense S-ODNs targeted to AFP genes inhibit the growth of human hepatoma cells and solid hepatoma, which is related to their cell apoptosis induction.展开更多
A major problem which is poorly understood in the management of bladder cancer is low sensitivity to chemotherapy and high recurrence after transurethral resection. Insulin-like growth factor 1 receptor (IGF-1R) signa...A major problem which is poorly understood in the management of bladder cancer is low sensitivity to chemotherapy and high recurrence after transurethral resection. Insulin-like growth factor 1 receptor (IGF-1R) signaling plays a very important role in progression, invasion and metastasis of bladder cancer cells. In this study, we investigated whether IGF-1R was involved in the growth stimulating activity and drug resistance of bladder cancer cells. The results showed: The mRNAs of IGF-1, IGF-2 and IGF-1R were strongly expressed in serum-free cultured T24 cell line, whereas normal urothelial cells did not express these factors/receptors or only in trace levels; T24 cell responded far better to growth stimulation by IGF-1 than did normal urothelial cells; blockage of IGF1R by antisense oligodeoxynucleotide (ODN) significantly inhibited the growth of T24 cell and enhanced sensitivity and apoptosis of T24 cells to mitomycin (MMC). These results suggested that blockage of IGF-IR signaling might potentially contribute to the treatment of bladder cancer cells which are insensitive to chemotherapy.展开更多
The chelates of metal ions with EDTA covalently linked to the 5′end of oligodeoxyribonuclotides(ODN),i.e,ODN5′EDTA·M(Ⅱ),are prepared,in which M(Ⅱ) is Fe(Ⅱ),Co(Ⅱ) or Cu(Ⅱ).The optimum pH value for forming t...The chelates of metal ions with EDTA covalently linked to the 5′end of oligodeoxyribonuclotides(ODN),i.e,ODN5′EDTA·M(Ⅱ),are prepared,in which M(Ⅱ) is Fe(Ⅱ),Co(Ⅱ) or Cu(Ⅱ).The optimum pH value for forming these three chelates is calculated.For ODN5′EDTA Fe(Ⅱ) pH value is 5.8 to 8.6,pH 4.6～8.1 for ODN5′EDTA Co(Ⅱ),and pH 3.4～5.7 for ODN5′EDTA Cu(Ⅱ).Under such pH value conditions neither can Mg(Ⅱ) ion,necessary for cleavage reaction,be competitive with Fe(Ⅱ),Co(Ⅱ) or Cu(Ⅱ) to form EDTA chelate,nor can it be precipitated.The cleavage mechanism of ODN5′EDTA Fe(Ⅱ) for DNA duplex is discussed.Modified ODN binds with DNA duplex in the major groove via hydrogen bond to form triple helix.In the presense of oxygen and reducing agent dithiothreitol,hydroxyl radicals species are generated as intermediates by catalysis of metal ions,and then oxidize the ribo ring and cut the doublestranded DNA at the sites close to the EDTA· Fe(Ⅱ).展开更多
基金Supported by the National Postdoctoral Science Foundation of China,No.199711.
文摘AIM: To evaluate antihepatoma effect of antisense phosphorothioate oligodeoxyribonucleotides (S-ODNs) targeted to alpha-fetoprotein (AFP) genes in vitro and in nude mice. METHODS: AFP gene expression was examined by immunocytochemical method or enzyme-linked immunosorbent assay. Effect of S-ODNs on SMMC-7721 human hepatoma cell growth in vitro was determined using microculture tetrazolium assay. In vitro antitumor activities of S-ODNs were monitored by measuring tumor weight differences in treated and control mice bearing SMMC-7721 xenografts. Induction of cell apoptosis was evaluated by fluorescence-activated cell sorter (FACS) analysis. RESULTS: Antisense S-ODN treatment led to reduced AFP gene expression. Specific antisense S-ODNs, but not control S-ODNs, inhibited the growth of hepatoma cells in vitro. In vitro, only antisense S-ODNs exhibited obvious antitumor activities. FACS analysis revealed that the growth inhibition by antisense S-ODNs was associated with their cell apoptosis induction. CONCLUSION: Antisense S-ODNs targeted to AFP genes inhibit the growth of human hepatoma cells and solid hepatoma, which is related to their cell apoptosis induction.
文摘A major problem which is poorly understood in the management of bladder cancer is low sensitivity to chemotherapy and high recurrence after transurethral resection. Insulin-like growth factor 1 receptor (IGF-1R) signaling plays a very important role in progression, invasion and metastasis of bladder cancer cells. In this study, we investigated whether IGF-1R was involved in the growth stimulating activity and drug resistance of bladder cancer cells. The results showed: The mRNAs of IGF-1, IGF-2 and IGF-1R were strongly expressed in serum-free cultured T24 cell line, whereas normal urothelial cells did not express these factors/receptors or only in trace levels; T24 cell responded far better to growth stimulation by IGF-1 than did normal urothelial cells; blockage of IGF1R by antisense oligodeoxynucleotide (ODN) significantly inhibited the growth of T24 cell and enhanced sensitivity and apoptosis of T24 cells to mitomycin (MMC). These results suggested that blockage of IGF-IR signaling might potentially contribute to the treatment of bladder cancer cells which are insensitive to chemotherapy.
文摘The chelates of metal ions with EDTA covalently linked to the 5′end of oligodeoxyribonuclotides(ODN),i.e,ODN5′EDTA·M(Ⅱ),are prepared,in which M(Ⅱ) is Fe(Ⅱ),Co(Ⅱ) or Cu(Ⅱ).The optimum pH value for forming these three chelates is calculated.For ODN5′EDTA Fe(Ⅱ) pH value is 5.8 to 8.6,pH 4.6～8.1 for ODN5′EDTA Co(Ⅱ),and pH 3.4～5.7 for ODN5′EDTA Cu(Ⅱ).Under such pH value conditions neither can Mg(Ⅱ) ion,necessary for cleavage reaction,be competitive with Fe(Ⅱ),Co(Ⅱ) or Cu(Ⅱ) to form EDTA chelate,nor can it be precipitated.The cleavage mechanism of ODN5′EDTA Fe(Ⅱ) for DNA duplex is discussed.Modified ODN binds with DNA duplex in the major groove via hydrogen bond to form triple helix.In the presense of oxygen and reducing agent dithiothreitol,hydroxyl radicals species are generated as intermediates by catalysis of metal ions,and then oxidize the ribo ring and cut the doublestranded DNA at the sites close to the EDTA· Fe(Ⅱ).
