Rad51 siRNA delivered by HVJ envelope vector enhances the anti‐cancer effect of cisplatin
Identifieur interne : 000A79 ( Istex/Checkpoint ); précédent : 000A78; suivant : 000A80Rad51 siRNA delivered by HVJ envelope vector enhances the anti‐cancer effect of cisplatin
Auteurs : Makoto Ito [Japon] ; Seiji Yamamoto [Japon] ; Keisuke Nimura [Japon] ; Kazuya Hiraoka [Japon] ; Katsuto Tamai [Japon] ; Yasufumi Kaneda [Japon]Source :
- The Journal of Gene Medicine [ 1099-498X ] ; 2005-08.
English descriptors
- Teeft :
- Antisense, Antisense oligonucleotides, Apoptosis, Biochem biophys, Cancer cell lines, Cancer cells, Cancer therapy, Cddp, Cddp conc, Cddp sensitivity, Cddp treatment, Cell lines, Cell number, Cisplatin, Copyright, Envelope vector, Hela, Hela cells, Hemagglutinating virus, Human cancer cells, Intraperitoneal injection, John wiley sons, Kaneda, Nhdf, Northern blot analysis, Oligonucleotides, Osaka university, Particle counter, Protein kinase, Protein level, Protein levels, Recombination, Relative cell count, Santa cruz, Scid mice, Similar results, Sirna, Sirna cddp, Sirna medium, Sirna transfer, Standard deviation, Synthetic sirna, Target protein, Transfection, Triplicate samples, Tumor growth, Tumor mass, Various concentrations, Western blot analysis.
Abstract
Background: Every cancer therapy appears to be transiently effective for cancer regression, but cancers gradually transform to be resistant to the therapy. Cancers also develop machineries to resist chemotherapy. Short interfering RNA (siRNA) has been evaluated as an attractive and effective tool for suppressing a target protein by specifically digesting its mRNA. Suppression of the machineries using siRNA may enhance the sensitivity to chemotherapy in cancers when combined with an effective delivery system. Methods: To enhance the anti‐cancer effect of chemotherapy, we transferred siRNA against Rad51 into various human cancer cells using the HVJ (hemagglutinating virus of Japan, Sendai virus) envelope vector in the presence or absence of cis‐diamminedichloroplatinum(II) (CDDP, cisplatin). The inhibition of cell growth was assessed by a modified MTT assay, counting cell number, or fluorescence‐activated cell sorting (FACS) analysis after Annexin V labeling. The synthetic Rad51 siRNA was also introduced into subcutaneous tumor masses of HeLa cells in SCID mice with or without intraperitoneal injection of CDDP, and tumor growth was monitored. Results: When synthetic Rad51 siRNA was delivered into HeLa cells using the HVJ envelope vector, no Rad51 transcripts were detected on day 2, and Rad51 protein completely disappeared for 4 days after siRNA transfer. When HeLa cells were incubated with 0.02 µg/ml CDDP for 3 h after siRNA transfer, the number of colonies decreased to approximately 10% of that with scrambled siRNA. The sensitivity to CDDP was enhanced in various human cancer cells, but not in normal human fibroblasts. When Rad51 siRNA was delivered into tumors using the HVJ envelope vector, the Rad51 transcript level was reduced to approximately 25%. Rad51 siRNA combined with CDDP significantly inhibited tumor growth when compared to siRNA or CDDP alone. Conclusions: Rad51 siRNA could enhance the sensitivity to CDDP in cancer cells both in vitro and in vivo. Our results suggest that the combination of CDDP and Rad51 siRNA will be an effective anti‐cancer protocol. Copyright © 2005 John Wiley & Sons, Ltd.
Url:
DOI: 10.1002/jgm.753
Affiliations:
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‐0871</wicri:noRegion></affiliation></author><author><name sortKey="Kaneda, Yasufumi" sort="Kaneda, Yasufumi" uniqKey="Kaneda Y" first="Yasufumi" last="Kaneda">Yasufumi Kaneda</name><affiliation wicri:level="1"><country xml:lang="fr">Japon</country><wicri:regionArea>Division of Gene Therapy Science, Graduate School of Medicine, Osaka University, 2‐2 Yamada‐oka, Suita, Osaka 565 ‐0871</wicri:regionArea><wicri:noRegion>Osaka 565 ‐0871</wicri:noRegion></affiliation><affiliation wicri:level="1"><country wicri:rule="url">Japon</country></affiliation><affiliation wicri:level="1"><country xml:lang="fr">Japon</country><wicri:regionArea>Correspondence address: Division of Gene Therapy Science, Graduate School of Medicine, Osaka University, 2‐2 Yamada‐oka, Suita, Osaka 565‐0871</wicri:regionArea><wicri:noRegion>Osaka 565‐0871</wicri:noRegion></affiliation></author></analytic><monogr></monogr><series><title level="j" type="main">The Journal of Gene Medicine</title><title level="j" type="alt">JOURNAL OF GENE MEDICINE, THE</title><idno type="ISSN">1099-498X</idno><idno type="eISSN">1521-2254</idno><imprint><biblScope unit="vol">7</biblScope><biblScope unit="issue">8</biblScope><biblScope unit="page" from="1044">1044</biblScope><biblScope unit="page" to="1052">1052</biblScope><biblScope unit="page-count">9</biblScope><publisher>John Wiley & Sons, Ltd.</publisher><pubPlace>Chichester, UK</pubPlace><date type="published" when="2005-08">2005-08</date></imprint><idno type="ISSN">1099-498X</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">1099-498X</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Antisense</term><term>Antisense oligonucleotides</term><term>Apoptosis</term><term>Biochem biophys</term><term>Cancer cell lines</term><term>Cancer cells</term><term>Cancer therapy</term><term>Cddp</term><term>Cddp conc</term><term>Cddp sensitivity</term><term>Cddp treatment</term><term>Cell lines</term><term>Cell number</term><term>Cisplatin</term><term>Copyright</term><term>Envelope vector</term><term>Hela</term><term>Hela cells</term><term>Hemagglutinating virus</term><term>Human cancer cells</term><term>Intraperitoneal injection</term><term>John wiley sons</term><term>Kaneda</term><term>Nhdf</term><term>Northern blot analysis</term><term>Oligonucleotides</term><term>Osaka university</term><term>Particle counter</term><term>Protein kinase</term><term>Protein level</term><term>Protein levels</term><term>Recombination</term><term>Relative cell count</term><term>Santa cruz</term><term>Scid mice</term><term>Similar results</term><term>Sirna</term><term>Sirna cddp</term><term>Sirna medium</term><term>Sirna transfer</term><term>Standard deviation</term><term>Synthetic sirna</term><term>Target protein</term><term>Transfection</term><term>Triplicate samples</term><term>Tumor growth</term><term>Tumor mass</term><term>Various concentrations</term><term>Western blot analysis</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Background: Every cancer therapy appears to be transiently effective for cancer regression, but cancers gradually transform to be resistant to the therapy. Cancers also develop machineries to resist chemotherapy. Short interfering RNA (siRNA) has been evaluated as an attractive and effective tool for suppressing a target protein by specifically digesting its mRNA. Suppression of the machineries using siRNA may enhance the sensitivity to chemotherapy in cancers when combined with an effective delivery system. Methods: To enhance the anti‐cancer effect of chemotherapy, we transferred siRNA against Rad51 into various human cancer cells using the HVJ (hemagglutinating virus of Japan, Sendai virus) envelope vector in the presence or absence of cis‐diamminedichloroplatinum(II) (CDDP, cisplatin). The inhibition of cell growth was assessed by a modified MTT assay, counting cell number, or fluorescence‐activated cell sorting (FACS) analysis after Annexin V labeling. The synthetic Rad51 siRNA was also introduced into subcutaneous tumor masses of HeLa cells in SCID mice with or without intraperitoneal injection of CDDP, and tumor growth was monitored. Results: When synthetic Rad51 siRNA was delivered into HeLa cells using the HVJ envelope vector, no Rad51 transcripts were detected on day 2, and Rad51 protein completely disappeared for 4 days after siRNA transfer. When HeLa cells were incubated with 0.02 µg/ml CDDP for 3 h after siRNA transfer, the number of colonies decreased to approximately 10% of that with scrambled siRNA. The sensitivity to CDDP was enhanced in various human cancer cells, but not in normal human fibroblasts. When Rad51 siRNA was delivered into tumors using the HVJ envelope vector, the Rad51 transcript level was reduced to approximately 25%. Rad51 siRNA combined with CDDP significantly inhibited tumor growth when compared to siRNA or CDDP alone. Conclusions: Rad51 siRNA could enhance the sensitivity to CDDP in cancer cells both in vitro and in vivo. Our results suggest that the combination of CDDP and Rad51 siRNA will be an effective anti‐cancer protocol. Copyright © 2005 John Wiley & Sons, Ltd.</div></front></TEI><affiliations><list><country><li>Japon</li></country></list><tree><country name="Japon"><noRegion><name sortKey="Ito, Makoto" sort="Ito, Makoto" uniqKey="Ito M" first="Makoto" last="Ito">Makoto Ito</name></noRegion><name sortKey="Hiraoka, Kazuya" sort="Hiraoka, Kazuya" uniqKey="Hiraoka K" first="Kazuya" last="Hiraoka">Kazuya Hiraoka</name><name sortKey="Kaneda, Yasufumi" sort="Kaneda, Yasufumi" uniqKey="Kaneda Y" first="Yasufumi" last="Kaneda">Yasufumi Kaneda</name><name sortKey="Kaneda, Yasufumi" sort="Kaneda, Yasufumi" uniqKey="Kaneda Y" first="Yasufumi" last="Kaneda">Yasufumi Kaneda</name><name sortKey="Kaneda, Yasufumi" sort="Kaneda, Yasufumi" uniqKey="Kaneda Y" first="Yasufumi" last="Kaneda">Yasufumi Kaneda</name><name sortKey="Nimura, Keisuke" sort="Nimura, Keisuke" uniqKey="Nimura K" first="Keisuke" last="Nimura">Keisuke Nimura</name><name sortKey="Tamai, Katsuto" sort="Tamai, Katsuto" uniqKey="Tamai K" first="Katsuto" last="Tamai">Katsuto Tamai</name><name sortKey="Yamamoto, Seiji" sort="Yamamoto, Seiji" uniqKey="Yamamoto S" first="Seiji" last="Yamamoto">Seiji Yamamoto</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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