DNA damage sensible engineered promoter for cellular biosensing of cytotoxicity
We have established a cytotoxic sensor cell line by transfecting HepG2 cells with a luciferase protein plasmid derived from the heat shock protein 70B' (HSP70B') promoter, which is induced by cytotoxic reagents. HSP70B genes are up-regulated by a wide-range of cytotoxic stimulators, in par...
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| Veröffentlicht in: | Biotechnology and bioengineering 2009-04, Vol.102 (5), p.1460-1465 |
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| creator | Wada, Ken-Ichi Hamaguchi, Yu Furukawa, Kiyoshi Taniguchi, Akiyoshi |
| description | We have established a cytotoxic sensor cell line by transfecting HepG2 cells with a luciferase protein plasmid derived from the heat shock protein 70B' (HSP70B') promoter, which is induced by cytotoxic reagents. HSP70B genes are up-regulated by a wide-range of cytotoxic stimulators, in particular, those that denature proteins. However, the HSP70B genes do not respond to DNA damage. We used a PCR array to detect marker genes of DNA damage-related cytotoxic stimulation and found the BTG2 gene to be one such gene. Analysis of the BTG2 gene functional promoter region by transfection of various deletion constructs into HepG2 cells indicated that the p53 and NFY biding sites on BTG2 are important for the response to DNA damage. We then constructed HepG2 sensor cells using the functional BTG2 promoter, and found that these sensor cells can specifically detect the cytotoxicity accompanied by DNA strand breaks with high sensitivity. Biotechnol. Bioeng. 2009;102: 1460-1465. |
| doi_str_mv | 10.1002/bit.22180 |
| format | Article |
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HSP70B genes are up-regulated by a wide-range of cytotoxic stimulators, in particular, those that denature proteins. However, the HSP70B genes do not respond to DNA damage. We used a PCR array to detect marker genes of DNA damage-related cytotoxic stimulation and found the BTG2 gene to be one such gene. Analysis of the BTG2 gene functional promoter region by transfection of various deletion constructs into HepG2 cells indicated that the p53 and NFY biding sites on BTG2 are important for the response to DNA damage. We then constructed HepG2 sensor cells using the functional BTG2 promoter, and found that these sensor cells can specifically detect the cytotoxicity accompanied by DNA strand breaks with high sensitivity. Biotechnol. Bioeng. 2009;102: 1460-1465.</description><identifier>ISSN: 0006-3592</identifier><identifier>EISSN: 1097-0290</identifier><identifier>DOI: 10.1002/bit.22180</identifier><identifier>PMID: 19031424</identifier><identifier>CODEN: BIBIAU</identifier><language>eng</language><publisher>Hoboken: Wiley Subscription Services, Inc., A Wiley Company</publisher><subject>Binding sites ; Biological and medical sciences ; Biosensing Techniques - methods ; biosensor ; Biosensors ; Biotechnology ; BTG2 ; Cell Line ; Cells ; Cells - drug effects ; Cytotoxins - toxicity ; DNA Damage ; Fundamental and applied biological sciences. Psychology ; Genes ; Genes, Reporter ; Genes, Tumor Suppressor ; Humans ; Immediate-Early Proteins - genetics ; Luciferases - genetics ; Luciferases - metabolism ; Methods. Procedures. Technologies ; Oligonucleotide Array Sequence Analysis ; p53 ; promoter ; Promoter Regions, Genetic ; Proteins ; Sensitivity and Specificity ; Toxicity ; Tumor Suppressor Proteins ; Various methods and equipments</subject><ispartof>Biotechnology and bioengineering, 2009-04, Vol.102 (5), p.1460-1465</ispartof><rights>Copyright © 2008 Wiley Periodicals, Inc.</rights><rights>2009 INIST-CNRS</rights><rights>Copyright John Wiley and Sons, Limited Apr 1, 2009</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c5700-f6ec6dacd40cf94956ea7e2ec886cc1361168bfadc56fdc492425807445ffcbb3</citedby><cites>FETCH-LOGICAL-c5700-f6ec6dacd40cf94956ea7e2ec886cc1361168bfadc56fdc492425807445ffcbb3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Fbit.22180$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fbit.22180$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,780,784,1417,27923,27924,45573,45574</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=21228079$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/19031424$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Wada, Ken-Ichi</creatorcontrib><creatorcontrib>Hamaguchi, Yu</creatorcontrib><creatorcontrib>Furukawa, Kiyoshi</creatorcontrib><creatorcontrib>Taniguchi, Akiyoshi</creatorcontrib><title>DNA damage sensible engineered promoter for cellular biosensing of cytotoxicity</title><title>Biotechnology and bioengineering</title><addtitle>Biotechnol. Bioeng</addtitle><description>We have established a cytotoxic sensor cell line by transfecting HepG2 cells with a luciferase protein plasmid derived from the heat shock protein 70B' (HSP70B') promoter, which is induced by cytotoxic reagents. HSP70B genes are up-regulated by a wide-range of cytotoxic stimulators, in particular, those that denature proteins. However, the HSP70B genes do not respond to DNA damage. We used a PCR array to detect marker genes of DNA damage-related cytotoxic stimulation and found the BTG2 gene to be one such gene. Analysis of the BTG2 gene functional promoter region by transfection of various deletion constructs into HepG2 cells indicated that the p53 and NFY biding sites on BTG2 are important for the response to DNA damage. We then constructed HepG2 sensor cells using the functional BTG2 promoter, and found that these sensor cells can specifically detect the cytotoxicity accompanied by DNA strand breaks with high sensitivity. Biotechnol. Bioeng. 2009;102: 1460-1465.</description><subject>Binding sites</subject><subject>Biological and medical sciences</subject><subject>Biosensing Techniques - methods</subject><subject>biosensor</subject><subject>Biosensors</subject><subject>Biotechnology</subject><subject>BTG2</subject><subject>Cell Line</subject><subject>Cells</subject><subject>Cells - drug effects</subject><subject>Cytotoxins - toxicity</subject><subject>DNA Damage</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genes</subject><subject>Genes, Reporter</subject><subject>Genes, Tumor Suppressor</subject><subject>Humans</subject><subject>Immediate-Early Proteins - genetics</subject><subject>Luciferases - genetics</subject><subject>Luciferases - metabolism</subject><subject>Methods. Procedures. Technologies</subject><subject>Oligonucleotide Array Sequence Analysis</subject><subject>p53</subject><subject>promoter</subject><subject>Promoter Regions, Genetic</subject><subject>Proteins</subject><subject>Sensitivity and Specificity</subject><subject>Toxicity</subject><subject>Tumor Suppressor Proteins</subject><subject>Various methods and equipments</subject><issn>0006-3592</issn><issn>1097-0290</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2009</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqF0U1rFDEYAOAgil2rB_-ADgUFD9PmeybH2motrNuDrQUvIZN5s6TOTGoyg91_b7azVhDEUwg87zdCLwk-JBjTo8aPh5SSGj9CC4JVVWKq8GO0wBjLkglF99CzlG7yt6qlfIr2iMKMcMoX6OJ0dVy0pjdrKBIMyTcdFDCs_QAQoS1uY-jDCLFwIRYWum7qTCwaH-7xsC6CK-xmDGO489aPm-foiTNdghe7dx9dffxwefKpXF6cnZ8cL0srKoxLJ8HK1tiWY-sUV0KCqYCCrWtpLWGSEFk3zrRWSNdariinosYV58I52zRsH72d8-YGf0yQRt37tO3PDBCmpBkjgtKa_xdSTKgSRGR48Be8CVMc8hCaElZJKvg227sZ2RhSiuD0bfS9iRtNsN7eQudb6PtbZPtql3Bqemj_yN3yM3izAyZZ07loBuvTg6MkD4Arld3R7H76Djb_rqjfn1_-Ll3OET6NcPcQYeJ3LStWCX29OtPX35Z0xU6_6s_Zv569M0GbdcxdXH3Jm2GYCCUrKdgvV5-4FQ</recordid><startdate>20090401</startdate><enddate>20090401</enddate><creator>Wada, Ken-Ichi</creator><creator>Hamaguchi, Yu</creator><creator>Furukawa, Kiyoshi</creator><creator>Taniguchi, Akiyoshi</creator><general>Wiley Subscription Services, Inc., A Wiley Company</general><general>Wiley</general><general>Wiley Subscription Services, Inc</general><scope>FBQ</scope><scope>BSCLL</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QF</scope><scope>7QO</scope><scope>7QQ</scope><scope>7SC</scope><scope>7SE</scope><scope>7SP</scope><scope>7SR</scope><scope>7T7</scope><scope>7TA</scope><scope>7TB</scope><scope>7U5</scope><scope>8BQ</scope><scope>8FD</scope><scope>C1K</scope><scope>F28</scope><scope>FR3</scope><scope>H8D</scope><scope>H8G</scope><scope>JG9</scope><scope>JQ2</scope><scope>KR7</scope><scope>L7M</scope><scope>L~C</scope><scope>L~D</scope><scope>P64</scope><scope>7TM</scope></search><sort><creationdate>20090401</creationdate><title>DNA damage sensible engineered promoter for cellular biosensing of cytotoxicity</title><author>Wada, Ken-Ichi ; Hamaguchi, Yu ; Furukawa, Kiyoshi ; Taniguchi, Akiyoshi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5700-f6ec6dacd40cf94956ea7e2ec886cc1361168bfadc56fdc492425807445ffcbb3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2009</creationdate><topic>Binding sites</topic><topic>Biological and medical sciences</topic><topic>Biosensing Techniques - methods</topic><topic>biosensor</topic><topic>Biosensors</topic><topic>Biotechnology</topic><topic>BTG2</topic><topic>Cell Line</topic><topic>Cells</topic><topic>Cells - drug effects</topic><topic>Cytotoxins - toxicity</topic><topic>DNA Damage</topic><topic>Fundamental and applied biological sciences. 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| subjects | Binding sites Biological and medical sciences Biosensing Techniques - methods biosensor Biosensors Biotechnology BTG2 Cell Line Cells Cells - drug effects Cytotoxins - toxicity DNA Damage Fundamental and applied biological sciences. Psychology Genes Genes, Reporter Genes, Tumor Suppressor Humans Immediate-Early Proteins - genetics Luciferases - genetics Luciferases - metabolism Methods. Procedures. Technologies Oligonucleotide Array Sequence Analysis p53 promoter Promoter Regions, Genetic Proteins Sensitivity and Specificity Toxicity Tumor Suppressor Proteins Various methods and equipments |
| title | DNA damage sensible engineered promoter for cellular biosensing of cytotoxicity |
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