Suppression of tumor growth by intratumoral injection of short hairpin RNA-expressing plasmid DNA targeting beta-catenin or hypoxia-inducible factor 1alpha.
To inhibit the growth of murine melanoma B16 cells in mice, we downregulated the gene expression of beta-catenin and hypoxia-inducible factor 1alpha (HIF1alpha) in the tumor cells by delivering short hairpin RNA (shRNA)-expressing plasmid DNA (pDNA) targeting one of these genes. Transfection of any of the shRNA-expressing pDNAs to B16 cells resulted in the reduction of the corresponding mRNA, which was associated with a reduced number of viable cells. A flow cytometric analysis of annexin V labeling assay was also performed to count the number of apoptotic cells. A flow cytometric analysis showed that the suppression of the expression of beta-catenin or HIF1alpha in B16 cells increased the number of apoptotic cells. An intratumoral injection of pshbeta-catenin (shRNA-expressing pDNA targeting beta-catenin) or pshHIF1alpha (shRNA-expressing pDNA targeting HIF1alpha) followed by electroporation greatly suppressed the expression of the corresponding target mRNA in the intradermal tumor tissue. The growth of the intradermal tumor was significantly (P<0.05) suppressed by the treatment. In conclusion, tumor growth was successfully inhibited by the intratumoral delivery of pshbeta-catenin or pshHIF1alpha.
Takahashi
Yuki
Y
Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto, Japan.
Nishikawa
Makiya
M
Takakura
Yoshinobu
Y
D000818:Animals; D017304:Annexin A5 / Q000235:genetics; D017209:Apoptosis / Q000187:drug effects; D004247:DNA / Q000235:genetics*; D018274:Electroporation; D005434:Flow Cytometry; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000096:biosynthesis* / Q000235:genetics*; D008297:Male; D008546:Melanoma, Experimental / Q000235:genetics / Q000628:therapy; D051379:Mice; D008810:Mice, Inbred C57BL; D009368:Neoplasm Transplantation; D009369:Neoplasms / Q000473:pathology / Q000628:therapy*; D009690:Nucleic Acid Conformation; D010957:Plasmids / Q000235:genetics*; D034622:RNA Interference / Q000187:drug effects; D016337:RNA, Catalytic / Q000235:genetics*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection; D051176:beta Catenin / Q000096:biosynthesis* / Q000235:genetics*
D017304:Annexin A5; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D016337:RNA, Catalytic; D012333:RNA, Messenger; D051176:beta Catenin; C418481:hairpin ribozyme; D004247:DNA