| 反义人DNMT1基因对乳腺癌细胞MDA-MB-435s增殖和细胞周期的影响 |
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| 引用本文： | 孙荣斌,鲁雅洁,魏钦俊.反义人DNMT1基因对乳腺癌细胞MDA-MB-435s增殖和细胞周期的影响[J].山西医科大学学报,2010,41(3):205-209,240. |
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| 作者姓名： | 孙荣斌 鲁雅洁 魏钦俊 |
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| 作者单位： | 1. 南京医科大学药学院,南京,210029
2. 南京医科大学基础医学院生物技术系 |
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| 基金项目： | 南京医科大学校基金资助项目 |
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| 摘 要： | 目的 构建表达人DNMT1基因(DNA methyltransferase 1,DNMT1)反义RNA的真核表达质粒,观察其对乳腺癌细胞MDA-MB-435s增殖的影响.方法 应用DNA重组技术,将人DNMT1基因的cDNA片段克隆至pcDNA3.1(-)载体中,构建DNMTl反义RNA真核表达质粒,测序验证,将其转染人人乳腺癌细胞系中,利用荧光定量PCR法检测转染前后细胞DN-MT1基因mRNA表达水平的变化;Western blot检测转染后DNMT1蛋白表达变化;应用MTT法检测细胞生长状况,绘制细胞生长曲线;流式细胞术分析细胞周期的变化.结果 成功构建出能够表达DNMT1反义RNA的真核表达质粒.转染该质粒后,与未转染组和转染正义质粒组相比,MDA-MB-435s细胞中DNMT1基因mRNA和DNMT1蛋白表达量均显著下降;细胞生长变慢;细胞周期分析可见S期明显减少,而G1/G0期细胞显著增加.结论 反义DNMT1能够特异性下调该基因的表达,并抑制乳腺癌细胞的增殖,为乳腺癌基因治疗提供了新的靶标.
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| 关 键 词： | DNMT1基因 反义RNA 乳腺癌 |
Effect of DNMT1 antisense RNA on proliferation and cell cycle of human breast cancer cell MDA-MB-435s |
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| SUN Rong-bing,LU Ya-jie,WEI Qin-jun.Effect of DNMT1 antisense RNA on proliferation and cell cycle of human breast cancer cell MDA-MB-435s[J].Journal of Shanxi Medical University,2010,41(3):205-209,240. |
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| Authors: | SUN Rong-bing LU Ya-jie WEI Qin-jun |
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| Institution: | SUN Rong-bing1,LU Ya-jie2,WEI Qin-jun2(1School of Pharmacy,Nanjing Medical University,Nanjing 210029,China,2Dept of Biotechnology,Nanjing Medical University) |
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| Abstract: | Objective To construct the antisense RNA expression vector targeting DNMT1 gene,and to investigate its effect on cell cycle,proliferation and apoptosis of breast cancer cell line MDA-MB-435s.Methods cDNA of DNMTl gene was inserted into pcDNA3.1(-)vector to construct antisense RNA recombinant plasmids by recombinant DNA technology.After transfection into breast cancer MDA-MB-435s cells,the mRNA expression levels of DNMT1 gene and protein were detected by real-time quantitative PCR and Results The recombinant... |
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| Keywords: | MDA-MB-435s |
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