ABSTRACT
Objective To investigate whether specific cellular uptake of 99Tcm-survivin-ASODN in nude mice bearing human HCC is influenced by its cytosine content.Methods Three kinds(A1,A2,A3) of synthesized survivin ASODN with three cytosine contents(10％,20％,30％),20 bases per single-strand were prepared.They were labeled with 99Tcm by conjugating with a bifunctional chelator HYNIC,purified through Cellufine GH-25 and then encapsulated with liposome.Antisense gene imaging and the biodistribution of 99Tcm-HYNIC-survivin ASODN in nude mice bearing HCC were performed.The data were analysed by Kruskal-Wallis H test.Results At 4 h post injection,all the 3 labelled compounds showed increased uptake by tumor,liver and kidney.With increase in cytosine content,the uptake increased in kidney (％ID/g:1.50±0.06,2.80±0.09 and 3.96±0.03),and decreased in tumor (％ID/g:2.08±0.08,1.69±0.01 and 1.20±0.09).T/NT in imaging (4.49-4.93,4.12-4.21,3.35-3.85;H=12.50,P＜0.05) and in biodistribution (4.08-4.94,4.02-4.18,3.66-3.85;H=10.82,P＜0.05) were all significantly different.Conclusion ASODN with lower cytosine content shows higher uptake by HCC tumor cells and less stasis in kidneys,thus providing better quality in antisense gene imaging.
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Objective: To investigate the effect of antisense oligodeoxynucleotide (ASODN) of Livin mRNA on the proliferation and apoptosis of human lung adenocarcinorna A549 cells. Methods: Livin ASODN was transfected into A549 cells mediated by cationic liposome. The proliferation rates of A549 cells were assessed by MTT method. The transcription of Livin mRNA was detected by RT-PCR. The Livin protein expression was determined by immunohistochemistry and confocal laser scanning microscopy before and after transfection. The apoptotic ratios of A549 cells were examined by acridine orange/ethidium bromide(AO/EB) fluorescent staining. Results: It was observed that A549 cells expressed both Livin α and Livin β simultaneously, and the distribution of Livin protein could be observed in both cytoplasm and nuclei. The proliferation of A549 cells was inhibited significantly by Livin ASODN in a dose-dependent manner (P<0.01). After transfection of Lip-ASODN at a final concentration of 400 nmol/L, the expressions of Uvin were inhibited at mRNA and protein levels and the apoptotic ratio reached (31.25 ± 5.75)%. The difference was significant compared with control group [ (3.23% ± 1.98)%, P<0.01]. Conclusion: Livin ASODN significantly down-regulates the expression of Livin mRNA and effectively inhibites the proliferation and induces the apoptosis of A549 cells. Therefore Livin gene may become a new target for gene therapy for lung cancer.
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Objective To investigate the effect of suppression of ischemia induced retinal neovascularization by VEGF antisense oligodeoxyribonucleotides. [WT5”HZ]Methods [WT5”BZ]Mouse models of hyperoxia induced ischemic retinopathy were established. Retrobulbar injections were performed with VEGF antisense oligodeoxyribonucleotides or NS in 4 groups:normal control and various doses respectively. The nuclei of new vessel buds extending from the retina into the vitreous in different groups were counted and compared under the light microscope. Results There were plenty of new vessel buds in the eyes of mice in hyperoxic condition., while the number of the nuclei of new vessel buds is less in the murine eyes with retrobulbar injection of VEGF antisense oligodeoxyribonucleotides,especially the nuclei were redused with 59.3% in eyes with large dose. Conclusion The proliferation of retinal new vessel may be suppressed by using the retrobulbar injection of VEGF antisense oligodeoxyribonucleotides.