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Article: Retrovirus-mediated delivery of HPV16 E7 antisense RNA inhibited tumorigenicity of CaSki cells

TitleRetrovirus-mediated delivery of HPV16 E7 antisense RNA inhibited tumorigenicity of CaSki cells
Authors
KeywordsAntisense
Gene therapy
HPV16
Retrovirus-mediated gene delivery
Issue Date2000
PublisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/locate/ygyno
Citation
Gynecologic Oncology, 2000, v. 78 n. 3 I, p. 293-301 How to Cite?
AbstractObjective. In cervical cancer, high-risk human papillomavirus (HPV) genes are expressed solely in cancerous cells and have been proposed to be the most important etiological factors for cervical cancer, thus making them suitable targets for gene therapy. In this study, we aim to inactivate the HPV16 E7 in CaSki cells and test the possibility of reducing the tumorigenicity of these cells. Methods. The full-length HPV16 E7 cDNA was cloned in the pBabe-puro or pWZL-Hygro retrovirus vector in reverse orientation and was stably transfected into CaSki cells by replication-defective retrovirus infection giving rise to CaSki-E7AS and CaSki-E7AS2X cells. Immunoprecipitation/Western analysis and real-time RT-PCR were performed to document the levels of HPV16 E7 gene product. Flow cytometry was performed to study changes in the cell cycle in response to reduced E7 protein. The expression of bcl-2, RB, and E2F-1 was studied using Western blot analysis. Tumorigenicity of CaSki, CaSki-E7AS, and CaSki-E7AS2X cells was assayed with subepidermal tumor growth in nude mice. Results. We have documented that the delivery of the antisense gene construct resulted in the reduction of HPV16 E7 protein expression and cell proliferation in CaSki cells. Furthermore, we demonstrated that these changes were accompanied by cell cycle arrest, up-regulation of RB, and down-regulation of E2F-1 and bc1-2 proteins. More importantly, dose-dependent transduction of the antisense HPV16E7 construct was able to inhibit and/or retard the tumorigenicity of CaSki cells in vivo. Conclusions. Down-regulation of HPV16 E7 with antisense RNA is beneficial in reducing the tumorigenicity of CaSki cells and can potentially be useful for HPV-associated malignancy gene therapy, (C) 2000 Academic Press.
Persistent Identifierhttp://hdl.handle.net/10722/77844
ISSN
2015 Impact Factor: 4.198
2015 SCImago Journal Rankings: 2.284
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorChoo, CKen_HK
dc.contributor.authorLing, MTen_HK
dc.contributor.authorSuen, CKMen_HK
dc.contributor.authorChan, KWen_HK
dc.contributor.authorKwong, YLen_HK
dc.date.accessioned2010-09-06T07:36:21Z-
dc.date.available2010-09-06T07:36:21Z-
dc.date.issued2000en_HK
dc.identifier.citationGynecologic Oncology, 2000, v. 78 n. 3 I, p. 293-301en_HK
dc.identifier.issn0090-8258en_HK
dc.identifier.urihttp://hdl.handle.net/10722/77844-
dc.description.abstractObjective. In cervical cancer, high-risk human papillomavirus (HPV) genes are expressed solely in cancerous cells and have been proposed to be the most important etiological factors for cervical cancer, thus making them suitable targets for gene therapy. In this study, we aim to inactivate the HPV16 E7 in CaSki cells and test the possibility of reducing the tumorigenicity of these cells. Methods. The full-length HPV16 E7 cDNA was cloned in the pBabe-puro or pWZL-Hygro retrovirus vector in reverse orientation and was stably transfected into CaSki cells by replication-defective retrovirus infection giving rise to CaSki-E7AS and CaSki-E7AS2X cells. Immunoprecipitation/Western analysis and real-time RT-PCR were performed to document the levels of HPV16 E7 gene product. Flow cytometry was performed to study changes in the cell cycle in response to reduced E7 protein. The expression of bcl-2, RB, and E2F-1 was studied using Western blot analysis. Tumorigenicity of CaSki, CaSki-E7AS, and CaSki-E7AS2X cells was assayed with subepidermal tumor growth in nude mice. Results. We have documented that the delivery of the antisense gene construct resulted in the reduction of HPV16 E7 protein expression and cell proliferation in CaSki cells. Furthermore, we demonstrated that these changes were accompanied by cell cycle arrest, up-regulation of RB, and down-regulation of E2F-1 and bc1-2 proteins. More importantly, dose-dependent transduction of the antisense HPV16E7 construct was able to inhibit and/or retard the tumorigenicity of CaSki cells in vivo. Conclusions. Down-regulation of HPV16 E7 with antisense RNA is beneficial in reducing the tumorigenicity of CaSki cells and can potentially be useful for HPV-associated malignancy gene therapy, (C) 2000 Academic Press.en_HK
dc.languageengen_HK
dc.publisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/locate/ygynoen_HK
dc.relation.ispartofGynecologic Oncologyen_HK
dc.subjectAntisenseen_HK
dc.subjectGene therapyen_HK
dc.subjectHPV16en_HK
dc.subjectRetrovirus-mediated gene deliveryen_HK
dc.subject.meshAnimalsen_HK
dc.subject.meshCarrier Proteinsen_HK
dc.subject.meshCell Cycle Proteinsen_HK
dc.subject.meshCell Differentiationen_HK
dc.subject.meshCell Divisionen_HK
dc.subject.meshDNA-Binding Proteinsen_HK
dc.subject.meshDown-Regulationen_HK
dc.subject.meshE2F Transcription Factorsen_HK
dc.subject.meshE2F1 Transcription Factoren_HK
dc.subject.meshFemaleen_HK
dc.subject.meshFlow Cytometryen_HK
dc.subject.meshGene Expression Regulation, Neoplasticen_HK
dc.subject.meshGene Expression Regulation, Viralen_HK
dc.subject.meshGene Silencingen_HK
dc.subject.meshHumansen_HK
dc.subject.meshMiceen_HK
dc.subject.meshMice, Inbred BALB Cen_HK
dc.subject.meshMice, Nudeen_HK
dc.subject.meshNeoplasm Transplantationen_HK
dc.subject.meshOncogene Proteins, Viral - antagonists & inhibitors - biosynthesis - geneticsen_HK
dc.subject.meshPapillomaviridae - geneticsen_HK
dc.subject.meshPapillomavirus E7 Proteinsen_HK
dc.subject.meshProto-Oncogene Proteins c-bcl-2 - biosynthesis - metabolismen_HK
dc.subject.meshRNA, Antisense - administration & dosage - geneticsen_HK
dc.subject.meshRetinoblastoma Protein - biosynthesis - metabolismen_HK
dc.subject.meshRetinoblastoma-Binding Protein 1en_HK
dc.subject.meshRetroviridae - geneticsen_HK
dc.subject.meshReverse Transcriptase Polymerase Chain Reactionen_HK
dc.subject.meshTranscription Factor DP1en_HK
dc.subject.meshTranscription Factors - biosynthesis - metabolismen_HK
dc.subject.meshTransfectionen_HK
dc.subject.meshTumor Cells, Cultureden_HK
dc.subject.meshUterine Cervical Neoplasms - pathology - therapy - virologyen_HK
dc.titleRetrovirus-mediated delivery of HPV16 E7 antisense RNA inhibited tumorigenicity of CaSki cellsen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0090-8258&volume=78&spage=293&epage=301&date=2000&atitle=Retrovirus-mediated+delivery+of+HPV16+E7+antisense+RNA+inhibited+tumorigenicity+of+CaSki+cellsen_HK
dc.identifier.emailLing, MT:patling@hkucc.hku.hken_HK
dc.identifier.emailChan, KW:hrmtckw@hku.hken_HK
dc.identifier.emailKwong, YL:ylkwong@hku.hken_HK
dc.identifier.authorityLing, MT=rp00449en_HK
dc.identifier.authorityChan, KW=rp00330en_HK
dc.identifier.authorityKwong, YL=rp00358en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1006/gyno.2000.5916en_HK
dc.identifier.pmid10985883-
dc.identifier.scopuseid_2-s2.0-0033812598en_HK
dc.identifier.hkuros56478en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0033812598&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume78en_HK
dc.identifier.issue3 Ien_HK
dc.identifier.spage293en_HK
dc.identifier.epage301en_HK
dc.identifier.isiWOS:000089317000005-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridChoo, CK=35866260100en_HK
dc.identifier.scopusauthoridLing, MT=7102229780en_HK
dc.identifier.scopusauthoridSuen, CKM=36766577900en_HK
dc.identifier.scopusauthoridChan, KW=16444133100en_HK
dc.identifier.scopusauthoridKwong, YL=7102818954en_HK

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