OBJECTIVE To determine whether Bc1-2 protein down-regulation can render NCI-460 cells more susceptible to gamma radiation-induced apoptosis by treatment with antisense oligonucleotide （ASODN） against the coding regi...OBJECTIVE To determine whether Bc1-2 protein down-regulation can render NCI-460 cells more susceptible to gamma radiation-induced apoptosis by treatment with antisense oligonucleotide （ASODN） against the coding region of Bcl-2 mRNA. METHODS Cell survival was determined using the trypan blue dye exclusion. Expression of the Bcl-2 protein was assayed using immunofluo- rescence labeling with fluoresce isothiocyanate. Apoptosis was determined by Giemsa staining and flow cytomertry. RESULTS It was found that Bcl-2 ASODN combined with radiation sig- nificantly reduced the number of viable cells （P〈0.05）. There was no difference in cell survival between a nonsense oligodeoxynucleotide/radiation combination and cells treated with radiation alone. Bcl-2 ASODN combined with radiation significantly inhibited expression of the Bcl-2 protein in the NCI-H460 cells （P〈0.05）. Using Giemsa staining, cells treated with Bcl-2 ASODN combined with radiation at 72 h displayed classic apoptotic changes. Apoptotic rates of the NCI-H460 cells treated with Bcl-2 ASODN combined with radiation significantly increased （P〈 0.05）, compared with either a nonsense oligodeoxynucleotide/radiation combination or radiation-treatment cells alone. CONCLUSION ASODN against the coding region of Bcl-2 mRNA increases radiation-induced apoptosis in NCI-H460 cells.展开更多
基金This work was supported by the Natural Science Program Foundation of the Guangdong Province (021195)Natural Science Program Foundation of the Guangdong Province (04010446).
文摘OBJECTIVE To determine whether Bc1-2 protein down-regulation can render NCI-460 cells more susceptible to gamma radiation-induced apoptosis by treatment with antisense oligonucleotide （ASODN） against the coding region of Bcl-2 mRNA. METHODS Cell survival was determined using the trypan blue dye exclusion. Expression of the Bcl-2 protein was assayed using immunofluo- rescence labeling with fluoresce isothiocyanate. Apoptosis was determined by Giemsa staining and flow cytomertry. RESULTS It was found that Bcl-2 ASODN combined with radiation sig- nificantly reduced the number of viable cells （P〈0.05）. There was no difference in cell survival between a nonsense oligodeoxynucleotide/radiation combination and cells treated with radiation alone. Bcl-2 ASODN combined with radiation significantly inhibited expression of the Bcl-2 protein in the NCI-H460 cells （P〈0.05）. Using Giemsa staining, cells treated with Bcl-2 ASODN combined with radiation at 72 h displayed classic apoptotic changes. Apoptotic rates of the NCI-H460 cells treated with Bcl-2 ASODN combined with radiation significantly increased （P〈 0.05）, compared with either a nonsense oligodeoxynucleotide/radiation combination or radiation-treatment cells alone. CONCLUSION ASODN against the coding region of Bcl-2 mRNA increases radiation-induced apoptosis in NCI-H460 cells.
