癌变·畸变·突变
癌變·畸變·突變
암변·기변·돌변
CARCINOGENSES,TERATOGENSIS AND MUTAGENESIS
2014年
2期
108-113
,共6页
邓婷婷%谢妮%黄艳华%黄海燕%李子刚%胡章立%袁建辉
鄧婷婷%謝妮%黃豔華%黃海燕%李子剛%鬍章立%袁建輝
산정정%사니%황염화%황해연%리자강%호장립%원건휘
毛细管电泳%DNA甲基化%MCF-7细胞%流式细胞术
毛細管電泳%DNA甲基化%MCF-7細胞%流式細胞術
모세관전영%DNA갑기화%MCF-7세포%류식세포술
capillary electrophoresis%DNA methylation%MCF-7 cell%flow cytometer
目的:建立对米托蒽醌(Mit)耐药的乳腺癌MCF-7细胞系,并探讨乳腺癌细胞产生耐药与其基因组甲基化水平之间的关系。方法:用0.005、0.010和0.020μmol/L浓度的米托蒽醌染毒乳腺癌MCF-7细胞,建立对米托蒽醌不同耐药程度的MCF-7耐药细胞系,采用流式细胞术检测各染毒组MCF-7细胞内药物荧光强度D(755)值,确定Mit耐药细胞系的建立;提取各组细胞的DNA,利用毛细管电泳法检测野生型对照组及各染毒组细胞全基因组DNA甲基化水平。结果:野生型对照组、0.005、0.010和0.020μmol/L米托蒽醌染毒组的(755)值依次为16.1±0.04、14.3±0.12、13.3±0.07和9.7±0.08,与野生型对照组相比,随着药物染毒浓度的增加,MCF-7细胞内药物的D(755)值逐渐减少,其中0.020μmol/L染毒组较对照组显著减少,差异有统计学意义(P<0.05),表明细胞耐药性增强;而上述各组DNA甲基化水平依次为42.25%±0.64%、37.97%±1.18%、34.27%±0.14%、31.16%±0.80%,与野生型对照组相比,各染毒组细胞基因组DNA甲基化水平逐渐降低(P<0.05),且各染毒组间两两比较差异均具有统计学意义(P均<0.05)。结论:成功建立了对Mit耐药的MCF-7细胞系;确定了毛细管电泳法检测基因组DNA甲基化水平的电泳分离条件。D
目的:建立對米託蒽醌(Mit)耐藥的乳腺癌MCF-7細胞繫,併探討乳腺癌細胞產生耐藥與其基因組甲基化水平之間的關繫。方法:用0.005、0.010和0.020μmol/L濃度的米託蒽醌染毒乳腺癌MCF-7細胞,建立對米託蒽醌不同耐藥程度的MCF-7耐藥細胞繫,採用流式細胞術檢測各染毒組MCF-7細胞內藥物熒光彊度D(755)值,確定Mit耐藥細胞繫的建立;提取各組細胞的DNA,利用毛細管電泳法檢測野生型對照組及各染毒組細胞全基因組DNA甲基化水平。結果:野生型對照組、0.005、0.010和0.020μmol/L米託蒽醌染毒組的(755)值依次為16.1±0.04、14.3±0.12、13.3±0.07和9.7±0.08,與野生型對照組相比,隨著藥物染毒濃度的增加,MCF-7細胞內藥物的D(755)值逐漸減少,其中0.020μmol/L染毒組較對照組顯著減少,差異有統計學意義(P<0.05),錶明細胞耐藥性增彊;而上述各組DNA甲基化水平依次為42.25%±0.64%、37.97%±1.18%、34.27%±0.14%、31.16%±0.80%,與野生型對照組相比,各染毒組細胞基因組DNA甲基化水平逐漸降低(P<0.05),且各染毒組間兩兩比較差異均具有統計學意義(P均<0.05)。結論:成功建立瞭對Mit耐藥的MCF-7細胞繫;確定瞭毛細管電泳法檢測基因組DNA甲基化水平的電泳分離條件。D
목적:건립대미탁은곤(Mit)내약적유선암MCF-7세포계,병탐토유선암세포산생내약여기기인조갑기화수평지간적관계。방법:용0.005、0.010화0.020μmol/L농도적미탁은곤염독유선암MCF-7세포,건립대미탁은곤불동내약정도적MCF-7내약세포계,채용류식세포술검측각염독조MCF-7세포내약물형광강도D(755)치,학정Mit내약세포계적건립;제취각조세포적DNA,이용모세관전영법검측야생형대조조급각염독조세포전기인조DNA갑기화수평。결과:야생형대조조、0.005、0.010화0.020μmol/L미탁은곤염독조적(755)치의차위16.1±0.04、14.3±0.12、13.3±0.07화9.7±0.08,여야생형대조조상비,수착약물염독농도적증가,MCF-7세포내약물적D(755)치축점감소,기중0.020μmol/L염독조교대조조현저감소,차이유통계학의의(P<0.05),표명세포내약성증강;이상술각조DNA갑기화수평의차위42.25%±0.64%、37.97%±1.18%、34.27%±0.14%、31.16%±0.80%,여야생형대조조상비,각염독조세포기인조DNA갑기화수평축점강저(P<0.05),차각염독조간량량비교차이균구유통계학의의(P균<0.05)。결론:성공건립료대Mit내약적MCF-7세포계;학정료모세관전영법검측기인조DNA갑기화수평적전영분리조건。D
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.