癌变·畸变·突变Issue(2)：108-113,6.DOI:10.3969/j.issn.1004-616x.2014.02.006
毛细管电泳法检测耐米托蒽醌乳腺癌细胞系全基因组甲基化水平
Determination of genomic DNA methylation levels in mitoxantrone-resistant breast cancer cell lines by capillary electrophoresis
摘要
Abstract
OBJECTIVE: To establish a mitoxantrone-resistant MCF-7 breast cancer cell line,and to explore the relationship between breast cancer cells developing drug resistance and their genomic methylation levels. METHODS:Using cells infected with 0,0.005,0.010 and 0.020 μmol/L mitoxantrone,different levels of drug resistance of MCF-7/Mit cell line was established. Flow cytometer was used to detect the accumulation of mitoxantrone in MCF-7 cells to confirm drug resistant cell lines. Then,we used capillary electrophoresis to evaluate genomic DNA methylation levels of wide type and cells with different degrees of resistance. RESULTS:Compared with the control group,with increasing mitoxantrone concentration,drug accumulation gradually decreased in MCF-7 cells. In the wild-type control group,0.005,0.010 and 0.020 μmol/L -treated groups the drug D(755) values were 16.1±0.04,14.3± 0.12,13.3±0.07 and 9.7±0.08,respectively. The 0.02 μmol/L exposure group was significantly reduced compared with the control group,indicating that drug resistance was significantly increased (P<0.05). In the wild-type control group,0.005,0.010 and 0.020μmol/L-treated groups,the DNA methylation levels were 42.25%±0.64%,37.97%±1.18%,34.27%±0.14% and 31.16%±0.80%, respectively. Compared with the control group, genomic DNA methylation levels in each group of treated cells gradually decreased (P<0.05),with significant differences between each two groups (P<0.05). CONCLUSION:Mit drug-resistant MCF-7 cell line was successfully set up. The separation condition of capillary electrophoresis when measuring genomic DNA methylation levels was determined.关键词
毛细管电泳/DNA甲基化/MCF-7细胞/流式细胞术Key words
capillary electrophoresis/DNA methylation/MCF-7 cell/flow cytometer分类
生物科学引用本文复制引用
邓婷婷,谢妮,黄艳华,黄海燕,李子刚,胡章立,袁建辉..毛细管电泳法检测耐米托蒽醌乳腺癌细胞系全基因组甲基化水平[J].癌变·畸变·突变,2014,(2):108-113,6.基金项目
国家自然科学基金(30500599)，广东省自然科学基金(9151503102000019)，深圳市科技计划重点项目(201201028)，深圳市科技研发资金项目(JCYJ20120615085512920；JCYJ20120613171430264；ZYC201006180477A)，深圳市孔雀计划(KQTD201103) （30500599）
