• 華中科技大學同濟醫(yī)學院附屬同濟醫(yī)院普通外科(武漢430030);;
  • 通訊作者: 鄒聲泉,Email: sqzou@tjh.tjmu.edu.cn;

目的觀察反義DNA甲基轉移酶3b(DNMT3b)基因真核表達載體轉染對人膽管癌QBC-939細胞中DNMT3b基因表達的影響。
方法構建反義DNMT3b基因真核表達載體,通過脂質體轉染到人膽管癌細胞株QBC-939中,G418篩選得到穩(wěn)定轉染細胞株,PCR鑒定外源性neoR基因在轉染細胞中的表達以確定轉染是否成功。半定量RT-PCR觀察轉染前后DNMT3b基因mRNA水平的變化,流式細胞術檢測轉染前后DNMT3b蛋白的變化。
結果反義DNMT3b基因真核表達載體轉染能使人膽管癌QBC-939細胞中DNMT3b基因的mRNA水平從0.956±0.053降低至0.209±0.023,差異有統(tǒng)計學意義(P<0.01); 流式細胞術檢測結果顯示,DNMT3b蛋白水平從(75.38±3.22)%降低到(29.87±3.46)%,差異有統(tǒng)計學意義(P<0.01)。
結論反義DNMT3b基因轉染能降低人膽管癌QBC-939細胞中DNMT3b基因的表達水平,為研究DNMT3b基因功能及其在膽管癌細胞中的作用提供了方法和實驗依據。

引用本文: 左石,郭偉,劉民鋒,董涇青,徐立寧,羅劍,鄒聲泉. 反義DNMT3b基因真核表達載體轉染對人膽管癌細胞DNMT3b基因表達的影響. 中國普外基礎與臨床雜志, 2006, 13(1): 56-60. doi: 復制

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  2. 2. Egger G, Liang G, Aparicio A, et al. Epigenetics in human disease and prospects for epigenetic therapy [J]. Nature, 2004; 429(6990)∶457.
  3. 3. Hermann A, Gowher H, Jeltsch A. Biochemistry and biology of mammalian DNA methyltransferases [J]. Cell Mol Life Sci, 2004; 61(19-20)∶2571.
  4. 4. Sambrook J, Russell DW, 黃培堂, 等譯. 分子克隆實驗指南 [M]. 第3版. 北京: 科學出版社, 2002∶629~630.
  5. 5. Jin F, Dowdy SC, Xiong Y, et al. Upregulation of DNA methyltransferase 3B expression in endometrial cancers [J]. Gynecol Oncol, 2005; 96(2)∶531.
  6. 6. Girault I, Tozlu S, Lidereau R, et al. Expression analysis of DNA methyltransferases 1, 3A, and 3B in sporadic breast carcinomas [J]. Clin Cancer Res, 2003; 9(12)∶4415.
  7. 7. Nagai M, Nakamura A, Makino R, et al. Expression of DNA (5cytosin)methyltransferases (DNMTs) in hepatocellular carcinomas [J]. Hepatol Res, 2003; 26(3)∶186.
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  12. 12. Lee LK, Roth CM. Antisense technology in molecular and cellular bioengineering [J]. Curr Opin Biotechnol, 2003; 14(5)∶505.