Anti-cell death engineering of CHO cells: Co-overexpression of Bcl-2 for apoptosis inhibition, Beclin-1 for autophagy induction

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dc.contributor.authorLee, Jae-Seongko
dc.contributor.authorHa, Tae-Kwangko
dc.contributor.authorPark, Jin-Hyoungko
dc.contributor.authorLee, Gyun-Minko
dc.date.accessioned2013-08-14T01:26:35Z-
dc.date.available2013-08-14T01:26:35Z-
dc.date.created2013-07-18-
dc.date.created2013-07-18-
dc.date.issued2013-08-
dc.identifier.citationBIOTECHNOLOGY AND BIOENGINEERING, v.110, no.8, pp.2195 - 2207-
dc.identifier.issn0006-3592-
dc.identifier.urihttp://hdl.handle.net/10203/175061-
dc.description.abstractGenetic engineering approaches to inhibit cell death in Chinese hamster ovary (CHO) cell cultures have been limited primarily to anti-apoptosis engineering. Recently, autophagy has received attention as a new anti-cell death engineering target in addition to apoptosis. In order to achieve a more efficient protection of cells from the stressful culture conditions, the simultaneous targeting of anti-apoptosis and pro-autophagy in CHO cells (DG44) was attempted by co-overexpressing an anti-apoptotic protein, Bcl-2, and a key regulator of autophagy pathway, Beclin-1, respectively. Co-overexpression of Bcl-2 and Beclin-1 exhibited a longer culture period as well as higher viability during serum-free suspension culture, compared with the control (without co-overexpression of Bcl-2 and Beclin-1) and Bcl-2 overexpression only. In addition to the efficient inhibition of apoptosis by Bcl-2 overexpression, Beclin-1 overexpression successfully induced the increase in the autophagic marker protein, LC3-II, and autophagosome formation with the decrease in mTOR activity. Co-immunoprecipitation and qRT-PCR experiments revealed that the enforced expression of Beclin-1 increased Ulk1 expression and level of free-Beclin-1 that did not bind to the Bcl-2 despite the Bcl-2 overexpression. Under other stressful culture conditions such as treatment with sodium butyrate and hyperosmolality, co-overexpression of Bcl-2 and Beclin-1 also protected the cells from cell death more efficiently than Bcl-2 overexpression only, implying the potential of autophagy induction. Taken together, the data obtained here provide the evidence that pro-autophagy engineering together with anti-apoptosis engineering yields a synergistic effect and successfully enhances the anti-cell death engineering of CHO cells. Biotechnol. Bioeng. 2013; 110: 2195-2207. (c) 2013 Wiley Periodicals, Inc.-
dc.languageEnglish-
dc.publisherWILEY-BLACKWELL-
dc.subjectHAMSTER OVARY CELLS-
dc.subjectREGULATES AUTOPHAGY-
dc.subjectSODIUM-BUTYRATE-
dc.subjectCULTURE-
dc.subjectPATHWAY-
dc.subjectGENES-
dc.subjectBATCH-
dc.subjectMACROAUTOPHAGY-
dc.subjectMETABOLISM-
dc.subjectEXPRESSION-
dc.titleAnti-cell death engineering of CHO cells: Co-overexpression of Bcl-2 for apoptosis inhibition, Beclin-1 for autophagy induction-
dc.typeArticle-
dc.identifier.wosid000320930800014-
dc.identifier.scopusid2-s2.0-84879605428-
dc.type.rimsART-
dc.citation.volume110-
dc.citation.issue8-
dc.citation.beginningpage2195-
dc.citation.endingpage2207-
dc.citation.publicationnameBIOTECHNOLOGY AND BIOENGINEERING-
dc.identifier.doi10.1002/bit.24879-
dc.contributor.localauthorLee, Gyun-Min-
dc.type.journalArticleArticle-
dc.subject.keywordAuthorBcl-2-
dc.subject.keywordAuthorBeclin-1-
dc.subject.keywordAuthorCHO-DG44-
dc.subject.keywordAuthorapoptosis-
dc.subject.keywordAuthorautophagy-
dc.subject.keywordPlusHAMSTER OVARY CELLS-
dc.subject.keywordPlusREGULATES AUTOPHAGY-
dc.subject.keywordPlusSODIUM-BUTYRATE-
dc.subject.keywordPlusCULTURE-
dc.subject.keywordPlusPATHWAY-
dc.subject.keywordPlusGENES-
dc.subject.keywordPlusBATCH-
dc.subject.keywordPlusMACROAUTOPHAGY-
dc.subject.keywordPlusMETABOLISM-
dc.subject.keywordPlusEXPRESSION-
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