Developing multidrug-resistant cells and exploring correlation between BCRP/ABCG2 over-expression and DNA methyltransferase
Expression of breast cancer resistance protein/ATP-binding cassette sub-family G member 2 （BCRP/ABCG2） is the major cause of chemotherapy failure. It is important to establish and characterize the multidrug resistance cells and to investigate the mechanism of multidrug resistance. Multidrug-resistan...
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| Veröffentlicht in: | Acta biochimica et biophysica Sinica 2010-12, Vol.42 (12), p.854-862 |
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| creator | Ji, Nana Yuan, Jianhui Liu, Jianjun Tian, Shengli |
| description | Expression of breast cancer resistance protein/ATP-binding cassette sub-family G member 2 （BCRP/ABCG2） is the major cause of chemotherapy failure. It is important to establish and characterize the multidrug resistance cells and to investigate the mechanism of multidrug resistance. Multidrug-resistant cells expressing BCRP/ABCG2 based on human breast cancer MCF-7/wt cells were developed by gradually increasing application of low concentration of mitoxantrone. Real-time quantitative PCR, western blot, and immunofluorescence assay were employed to analyze BCRP mRNA and protein expression. Drug accumulation in the cells was measured by flow cytometry and DNA methyltransferases were analyzed by western blot. The results indicated that the inhibitory ratio of cell proliferative growth exhibited an exponential relation with the concentration of mitoxantrone. The ICs0 of MCF-7/wt cells to mitoxantrone was found to be 0.42 μM. 3-（4,5-Dimethylthlthiazol-2-YI）- 2,5-Diphenyltetrazolium Bromide assay indicated that the mitoxantrone-resistant cells at different stages exhibited cross-resistance to adriamycin and taxol. BCRP/ABCG2 mRNA and protein levels in the mitoxantrone-resistant cells at different stages increased with increasing concentration of mitoxantrone. Intracellular accumulation of mitoxantrone in the cells decreased with the increase of the BCRP/ABCG2 expression levels. DNA methyltransferase 1 （DNMT1） and DNA methyltransferase 3a （DNMT3a） expressions in the cells at different stages decreased slightly, whereas DNA methyltransferase 3b （DNMT3b） expression decreases significantly. BCRP/ABCG2 overexpression and its drug-efflux function in the drug-resistant cells are the main factors to produce multidrug resistance. Our results suggest that multidrug resistance is related to overexpression of BCRP/ ABCG2 and the decrease of DNA methyltransferases, especially DNMT3b. |
| doi_str_mv | 10.1093/abbs/gmq097 |
| format | Article |
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It is important to establish and characterize the multidrug resistance cells and to investigate the mechanism of multidrug resistance. Multidrug-resistant cells expressing BCRP/ABCG2 based on human breast cancer MCF-7/wt cells were developed by gradually increasing application of low concentration of mitoxantrone. Real-time quantitative PCR, western blot, and immunofluorescence assay were employed to analyze BCRP mRNA and protein expression. Drug accumulation in the cells was measured by flow cytometry and DNA methyltransferases were analyzed by western blot. The results indicated that the inhibitory ratio of cell proliferative growth exhibited an exponential relation with the concentration of mitoxantrone. The ICs0 of MCF-7/wt cells to mitoxantrone was found to be 0.42 μM. 3-（4,5-Dimethylthlthiazol-2-YI）- 2,5-Diphenyltetrazolium Bromide assay indicated that the mitoxantrone-resistant cells at different stages exhibited cross-resistance to adriamycin and taxol. BCRP/ABCG2 mRNA and protein levels in the mitoxantrone-resistant cells at different stages increased with increasing concentration of mitoxantrone. Intracellular accumulation of mitoxantrone in the cells decreased with the increase of the BCRP/ABCG2 expression levels. DNA methyltransferase 1 （DNMT1） and DNA methyltransferase 3a （DNMT3a） expressions in the cells at different stages decreased slightly, whereas DNA methyltransferase 3b （DNMT3b） expression decreases significantly. BCRP/ABCG2 overexpression and its drug-efflux function in the drug-resistant cells are the main factors to produce multidrug resistance. Our results suggest that multidrug resistance is related to overexpression of BCRP/ ABCG2 and the decrease of DNA methyltransferases, especially DNMT3b.</description><identifier>ISSN: 1672-9145</identifier><identifier>EISSN: 1745-7270</identifier><identifier>DOI: 10.1093/abbs/gmq097</identifier><identifier>PMID: 21106767</identifier><language>eng</language><publisher>China: Oxford University Press</publisher><subject>Antineoplastic Agents - pharmacology ; ATP Binding Cassette Transporter, Sub-Family G, Member 2 ; ATP-Binding Cassette Transporters - antagonists & inhibitors ; ATP-Binding Cassette Transporters - genetics ; ATP-Binding Cassette Transporters - metabolism ; Blotting, Western ; Breast Neoplasms - metabolism ; Breast Neoplasms - pathology ; Cell Proliferation - drug effects ; DNA Modification Methylases - genetics ; DNA Modification Methylases - metabolism ; DNA甲基转移酶 ; Drug Resistance, Neoplasm ; Female ; Fluorescent Antibody Technique ; Gene Expression Regulation, Neoplastic - drug effects ; Humans ; Mycotoxins - pharmacology ; Neoplasm Proteins - genetics ; Neoplasm Proteins - metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Cells, Cultured ; 人乳腺癌细胞 ; 多药耐药 ; 实时定量PCR ; 米托蒽醌 ; 蛋白表达 ; 过度表达</subject><ispartof>Acta biochimica et biophysica Sinica, 2010-12, Vol.42 (12), p.854-862</ispartof><rights>The Author 2010. Published by ABBS Editorial Office in association with Oxford University Press on behalf of the Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences. 2010</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c388t-d78d1fd613f6d4e1713119bf1d31bb211fa34941969f3356e1d1f975dd7c950b3</citedby><cites>FETCH-LOGICAL-c388t-d78d1fd613f6d4e1713119bf1d31bb211fa34941969f3356e1d1f975dd7c950b3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://image.cqvip.com/vip1000/qk/90160X/90160X.jpg</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21106767$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ji, Nana</creatorcontrib><creatorcontrib>Yuan, Jianhui</creatorcontrib><creatorcontrib>Liu, Jianjun</creatorcontrib><creatorcontrib>Tian, Shengli</creatorcontrib><title>Developing multidrug-resistant cells and exploring correlation between BCRP/ABCG2 over-expression and DNA methyltransferase</title><title>Acta biochimica et biophysica Sinica</title><addtitle>Acta Biochimica et Biophysica Sinica</addtitle><description>Expression of breast cancer resistance protein/ATP-binding cassette sub-family G member 2 （BCRP/ABCG2） is the major cause of chemotherapy failure. It is important to establish and characterize the multidrug resistance cells and to investigate the mechanism of multidrug resistance. Multidrug-resistant cells expressing BCRP/ABCG2 based on human breast cancer MCF-7/wt cells were developed by gradually increasing application of low concentration of mitoxantrone. Real-time quantitative PCR, western blot, and immunofluorescence assay were employed to analyze BCRP mRNA and protein expression. Drug accumulation in the cells was measured by flow cytometry and DNA methyltransferases were analyzed by western blot. The results indicated that the inhibitory ratio of cell proliferative growth exhibited an exponential relation with the concentration of mitoxantrone. The ICs0 of MCF-7/wt cells to mitoxantrone was found to be 0.42 μM. 3-（4,5-Dimethylthlthiazol-2-YI）- 2,5-Diphenyltetrazolium Bromide assay indicated that the mitoxantrone-resistant cells at different stages exhibited cross-resistance to adriamycin and taxol. BCRP/ABCG2 mRNA and protein levels in the mitoxantrone-resistant cells at different stages increased with increasing concentration of mitoxantrone. Intracellular accumulation of mitoxantrone in the cells decreased with the increase of the BCRP/ABCG2 expression levels. DNA methyltransferase 1 （DNMT1） and DNA methyltransferase 3a （DNMT3a） expressions in the cells at different stages decreased slightly, whereas DNA methyltransferase 3b （DNMT3b） expression decreases significantly. BCRP/ABCG2 overexpression and its drug-efflux function in the drug-resistant cells are the main factors to produce multidrug resistance. Our results suggest that multidrug resistance is related to overexpression of BCRP/ ABCG2 and the decrease of DNA methyltransferases, especially DNMT3b.</description><subject>Antineoplastic Agents - pharmacology</subject><subject>ATP Binding Cassette Transporter, Sub-Family G, Member 2</subject><subject>ATP-Binding Cassette Transporters - antagonists & inhibitors</subject><subject>ATP-Binding Cassette Transporters - genetics</subject><subject>ATP-Binding Cassette Transporters - metabolism</subject><subject>Blotting, Western</subject><subject>Breast Neoplasms - metabolism</subject><subject>Breast Neoplasms - pathology</subject><subject>Cell Proliferation - drug effects</subject><subject>DNA Modification Methylases - genetics</subject><subject>DNA Modification Methylases - metabolism</subject><subject>DNA甲基转移酶</subject><subject>Drug Resistance, Neoplasm</subject><subject>Female</subject><subject>Fluorescent Antibody Technique</subject><subject>Gene Expression Regulation, Neoplastic - drug effects</subject><subject>Humans</subject><subject>Mycotoxins - pharmacology</subject><subject>Neoplasm Proteins - genetics</subject><subject>Neoplasm Proteins - metabolism</subject><subject>Reverse Transcriptase Polymerase Chain Reaction</subject><subject>Tumor Cells, Cultured</subject><subject>人乳腺癌细胞</subject><subject>多药耐药</subject><subject>实时定量PCR</subject><subject>米托蒽醌</subject><subject>蛋白表达</subject><subject>过度表达</subject><issn>1672-9145</issn><issn>1745-7270</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqF0E1v1DAQBmALgegHnLijiEM5oLCeOLHXx-22FKQKEIKz5cTjbcCJs7ZTqPjzONqFK6fx4fE7mpeQF0DfApVspds2rnbDnkrxiJyCqJtSVII-zm8uqlJC3ZyQsxi_U8o4B_qUnFQAlAsuTsnvK7xH56d-3BXD7FJvwrwrA8Y-Jj2mokPnYqFHU-CvyfmwuM6HgE6n3o9Fi-kn4lhcbr98Xm0utzdV4e8xlFnnkLiQ5fPVx00xYLp7cCnoMVoMOuIz8sRqF_H5cZ6Tb--uv27fl7efbj5sN7dlx9brVBqxNmANB2a5qREEMADZWjAM2jafYjWrZQ2SS8tYwxEyl6IxRnSyoS07J68PuVPw-xljUkMfl8P0iH6Oag11zaumolm-Ocgu-BgDWjWFftDhQQFVS9lqKVsdys765TF3bgc0_-zfdjO4OAA_T_9JenXce-fH3T6XrFrd_bC9Q8U4o01NBfsDb_eVrA</recordid><startdate>20101201</startdate><enddate>20101201</enddate><creator>Ji, Nana</creator><creator>Yuan, Jianhui</creator><creator>Liu, Jianjun</creator><creator>Tian, Shengli</creator><general>Oxford University Press</general><scope>2RA</scope><scope>92L</scope><scope>CQIGP</scope><scope>W94</scope><scope>WU4</scope><scope>~WA</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>7X8</scope></search><sort><creationdate>20101201</creationdate><title>Developing multidrug-resistant cells and exploring correlation between BCRP/ABCG2 over-expression and DNA methyltransferase</title><author>Ji, Nana ; Yuan, Jianhui ; Liu, Jianjun ; Tian, Shengli</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c388t-d78d1fd613f6d4e1713119bf1d31bb211fa34941969f3356e1d1f975dd7c950b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>Antineoplastic Agents - pharmacology</topic><topic>ATP Binding Cassette Transporter, Sub-Family G, Member 2</topic><topic>ATP-Binding Cassette Transporters - antagonists & inhibitors</topic><topic>ATP-Binding Cassette Transporters - genetics</topic><topic>ATP-Binding Cassette Transporters - metabolism</topic><topic>Blotting, Western</topic><topic>Breast Neoplasms - metabolism</topic><topic>Breast Neoplasms - pathology</topic><topic>Cell Proliferation - drug effects</topic><topic>DNA Modification Methylases - genetics</topic><topic>DNA Modification Methylases - metabolism</topic><topic>DNA甲基转移酶</topic><topic>Drug Resistance, Neoplasm</topic><topic>Female</topic><topic>Fluorescent Antibody Technique</topic><topic>Gene Expression Regulation, Neoplastic - drug effects</topic><topic>Humans</topic><topic>Mycotoxins - pharmacology</topic><topic>Neoplasm Proteins - genetics</topic><topic>Neoplasm Proteins - metabolism</topic><topic>Reverse Transcriptase Polymerase Chain Reaction</topic><topic>Tumor Cells, Cultured</topic><topic>人乳腺癌细胞</topic><topic>多药耐药</topic><topic>实时定量PCR</topic><topic>米托蒽醌</topic><topic>蛋白表达</topic><topic>过度表达</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ji, Nana</creatorcontrib><creatorcontrib>Yuan, Jianhui</creatorcontrib><creatorcontrib>Liu, Jianjun</creatorcontrib><creatorcontrib>Tian, Shengli</creatorcontrib><collection>中文科技期刊数据库</collection><collection>中文科技期刊数据库-CALIS站点</collection><collection>中文科技期刊数据库-7.0平台</collection><collection>中文科技期刊数据库-自然科学</collection><collection>中文科技期刊数据库-自然科学-生物科学</collection><collection>中文科技期刊数据库- 镜像站点</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Acta biochimica et biophysica Sinica</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ji, Nana</au><au>Yuan, Jianhui</au><au>Liu, Jianjun</au><au>Tian, Shengli</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Developing multidrug-resistant cells and exploring correlation between BCRP/ABCG2 over-expression and DNA methyltransferase</atitle><jtitle>Acta biochimica et biophysica Sinica</jtitle><addtitle>Acta Biochimica et Biophysica Sinica</addtitle><date>2010-12-01</date><risdate>2010</risdate><volume>42</volume><issue>12</issue><spage>854</spage><epage>862</epage><pages>854-862</pages><issn>1672-9145</issn><eissn>1745-7270</eissn><abstract>Expression of breast cancer resistance protein/ATP-binding cassette sub-family G member 2 （BCRP/ABCG2） is the major cause of chemotherapy failure. It is important to establish and characterize the multidrug resistance cells and to investigate the mechanism of multidrug resistance. Multidrug-resistant cells expressing BCRP/ABCG2 based on human breast cancer MCF-7/wt cells were developed by gradually increasing application of low concentration of mitoxantrone. Real-time quantitative PCR, western blot, and immunofluorescence assay were employed to analyze BCRP mRNA and protein expression. Drug accumulation in the cells was measured by flow cytometry and DNA methyltransferases were analyzed by western blot. The results indicated that the inhibitory ratio of cell proliferative growth exhibited an exponential relation with the concentration of mitoxantrone. The ICs0 of MCF-7/wt cells to mitoxantrone was found to be 0.42 μM. 3-（4,5-Dimethylthlthiazol-2-YI）- 2,5-Diphenyltetrazolium Bromide assay indicated that the mitoxantrone-resistant cells at different stages exhibited cross-resistance to adriamycin and taxol. BCRP/ABCG2 mRNA and protein levels in the mitoxantrone-resistant cells at different stages increased with increasing concentration of mitoxantrone. Intracellular accumulation of mitoxantrone in the cells decreased with the increase of the BCRP/ABCG2 expression levels. DNA methyltransferase 1 （DNMT1） and DNA methyltransferase 3a （DNMT3a） expressions in the cells at different stages decreased slightly, whereas DNA methyltransferase 3b （DNMT3b） expression decreases significantly. BCRP/ABCG2 overexpression and its drug-efflux function in the drug-resistant cells are the main factors to produce multidrug resistance. Our results suggest that multidrug resistance is related to overexpression of BCRP/ ABCG2 and the decrease of DNA methyltransferases, especially DNMT3b.</abstract><cop>China</cop><pub>Oxford University Press</pub><pmid>21106767</pmid><doi>10.1093/abbs/gmq097</doi><tpages>9</tpages></addata></record> |
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| subjects | Antineoplastic Agents - pharmacology ATP Binding Cassette Transporter, Sub-Family G, Member 2 ATP-Binding Cassette Transporters - antagonists & inhibitors ATP-Binding Cassette Transporters - genetics ATP-Binding Cassette Transporters - metabolism Blotting, Western Breast Neoplasms - metabolism Breast Neoplasms - pathology Cell Proliferation - drug effects DNA Modification Methylases - genetics DNA Modification Methylases - metabolism DNA甲基转移酶 Drug Resistance, Neoplasm Female Fluorescent Antibody Technique Gene Expression Regulation, Neoplastic - drug effects Humans Mycotoxins - pharmacology Neoplasm Proteins - genetics Neoplasm Proteins - metabolism Reverse Transcriptase Polymerase Chain Reaction Tumor Cells, Cultured 人乳腺癌细胞 多药耐药 实时定量PCR 米托蒽醌 蛋白表达 过度表达 |
| title | Developing multidrug-resistant cells and exploring correlation between BCRP/ABCG2 over-expression and DNA methyltransferase |
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