Objective： To explore and investigate the selection of effective antisense oligodeoxynuleotides with the help of computer and RNAstructure folding software. Methods： Bcl-2 gene was used as the target gene and five a...Objective： To explore and investigate the selection of effective antisense oligodeoxynuleotides with the help of computer and RNAstructure folding software. Methods： Bcl-2 gene was used as the target gene and five antisense oligodeoxynuleotides were designed to be bound to Bcl-2 mRNA optimal secondary structure regions that were predicted free from intramolecular fold or instability of free energy. The five antisense oligodeoxynucleotides were studied with experimental assay of leukemia cells, including cell grow assay with tropan blue exclusion, expression of Bcl-2 protein detected with immunochemistry and flowcytometry, Bcl-2 mRNA content detected with RT-PCR technique, as well as apoptosis observed and determined with morphonological method, electrophoresis and flowcytometry. Results： The results showed that two of the five antisense oligodeoxynucleotides were effective antisense oligodeoxynucleotides, which were able to inhibit cell growth in leukemia, to decrease the level of Bcl-2 mRNA and protein, to induce apoptosis of leukemia cells significantly. Conclusion： The computational prediction of antisense efficacy is faster than other methods and more efficient, which can potentially speed the development of sequences for both research and clinical applications.展开更多
Objective To explore the effects of liposome C erbB 2 antisense phosphorothioate oligodeoxynucleotides (S ODNs) on C erbB 2 proto oncogene expression and cell proliferation in human ovarian cancer cells Metho...Objective To explore the effects of liposome C erbB 2 antisense phosphorothioate oligodeoxynucleotides (S ODNs) on C erbB 2 proto oncogene expression and cell proliferation in human ovarian cancer cells Methods The effects of liposome C erbB 2 S ODNs on C erbB 2 protein expression, cell cycle and cell proliferation in human ovarian cancer cells were studied by means of flow cytometry and 3H thymidine incorporation Results Liposome C erbB 2 S ODNs can specifically reduce C erbB 2 protein expression in human ovarian cancer cells, accompanied by a 30% inhibition of cell proliferation The effectiveness of liposome C erbB 2 S ODNs on the expression of C erbB 2 was about 40 times higher than that of C erbB 2 S ODNs Conclusions The data suggest that antisense therapy might be a useful method of gene therapy in ovarian cancer The effectiveness of C erbB 2 S ODNs could be greatly increased by adsorption of S ODNs by liposomes展开更多
Objective: To explore the effect of homeobox B2 (HOXB2) antisense oligodeoxynucleotides (asodn) on the proliferation and expression of primary human umbilical vein endothelial cells (HUVECs). Methods: Various concentr...Objective: To explore the effect of homeobox B2 (HOXB2) antisense oligodeoxynucleotides (asodn) on the proliferation and expression of primary human umbilical vein endothelial cells (HUVECs). Methods: Various concentrations of HOXB2 asodn modified by thiophosphate transfected the induction of liposome into HUVECs. MTT and RT-PCR methods were employed to determine the effect of different concentrations of asodn on the endothelial proliferation and the expression level of HOXB2 mRNA. Results: After the transfection of HOXB2 asodn, the endothelial proliferation was inhibited in a dose-dependent fashion. Simultaneously, the expression of HOXB2 mRNA decreased significantly. Conclusions: HOXB2 plays an important role in the proliferation of endothelia.展开更多
文摘Objective： To explore and investigate the selection of effective antisense oligodeoxynuleotides with the help of computer and RNAstructure folding software. Methods： Bcl-2 gene was used as the target gene and five antisense oligodeoxynuleotides were designed to be bound to Bcl-2 mRNA optimal secondary structure regions that were predicted free from intramolecular fold or instability of free energy. The five antisense oligodeoxynucleotides were studied with experimental assay of leukemia cells, including cell grow assay with tropan blue exclusion, expression of Bcl-2 protein detected with immunochemistry and flowcytometry, Bcl-2 mRNA content detected with RT-PCR technique, as well as apoptosis observed and determined with morphonological method, electrophoresis and flowcytometry. Results： The results showed that two of the five antisense oligodeoxynucleotides were effective antisense oligodeoxynucleotides, which were able to inhibit cell growth in leukemia, to decrease the level of Bcl-2 mRNA and protein, to induce apoptosis of leukemia cells significantly. Conclusion： The computational prediction of antisense efficacy is faster than other methods and more efficient, which can potentially speed the development of sequences for both research and clinical applications.
基金ThisworkwassupportedbytheNationalNaturalScienceFoundationof China (No 39470 72 4)
文摘Objective To explore the effects of liposome C erbB 2 antisense phosphorothioate oligodeoxynucleotides (S ODNs) on C erbB 2 proto oncogene expression and cell proliferation in human ovarian cancer cells Methods The effects of liposome C erbB 2 S ODNs on C erbB 2 protein expression, cell cycle and cell proliferation in human ovarian cancer cells were studied by means of flow cytometry and 3H thymidine incorporation Results Liposome C erbB 2 S ODNs can specifically reduce C erbB 2 protein expression in human ovarian cancer cells, accompanied by a 30% inhibition of cell proliferation The effectiveness of liposome C erbB 2 S ODNs on the expression of C erbB 2 was about 40 times higher than that of C erbB 2 S ODNs Conclusions The data suggest that antisense therapy might be a useful method of gene therapy in ovarian cancer The effectiveness of C erbB 2 S ODNs could be greatly increased by adsorption of S ODNs by liposomes
基金theNationalKeyBasicResearchProjectofChina (No .G19990 5 42 0 5 )
文摘Objective: To explore the effect of homeobox B2 (HOXB2) antisense oligodeoxynucleotides (asodn) on the proliferation and expression of primary human umbilical vein endothelial cells (HUVECs). Methods: Various concentrations of HOXB2 asodn modified by thiophosphate transfected the induction of liposome into HUVECs. MTT and RT-PCR methods were employed to determine the effect of different concentrations of asodn on the endothelial proliferation and the expression level of HOXB2 mRNA. Results: After the transfection of HOXB2 asodn, the endothelial proliferation was inhibited in a dose-dependent fashion. Simultaneously, the expression of HOXB2 mRNA decreased significantly. Conclusions: HOXB2 plays an important role in the proliferation of endothelia.
