【摘要】： 【研究目的】 本文探討CK13基因?qū)Ρ茄拾〩NE1細胞株放射敏感性的影響。我們采用脂質(zhì)體介導轉(zhuǎn)染方法將攜帶CK13基因的真核表達質(zhì)粒CK13-A和CK13-B,空載體質(zhì)粒pEGFP-N1分別轉(zhuǎn)染到鼻咽癌細胞株HNE1中。克隆形成實驗測定細胞存活分數(shù)。采用單擊多靶模型和線性二次函數(shù)模型擬合細胞存活曲線,求出放射生物學參數(shù)D0、Dq、N和α、β、α/β、SF2值,探討CK13基因與鼻咽癌HNE1細胞株放射敏感性的關系,以CK13基因為靶點進行相關研究,可為臨床提高鼻咽癌放療敏感性提供新的思路。 【研究方法】 1.目的基因的制備;重組DNA;轉(zhuǎn)染;篩選。 2.攜帶CK13基因的質(zhì)粒CK13-A、CK13-B和空載體質(zhì)粒(pEGFE-N1)的轉(zhuǎn)化、擴增、酶切與測序鑒定分析。 3.陽離子脂質(zhì)體Lipofectamine~(TM)2000介導,將CK13-A;CK13-B質(zhì)粒和空載體質(zhì)粒pEGFP-N1轉(zhuǎn)染至HNE1細胞中。分別命名為A組、B組、O組和H組。 4.克隆形成實驗測定細胞劑量存活分數(shù),采用線性二次函數(shù)模型(L-Q模型)和單擊多靶模型擬合細胞輻射劑量存活曲線,分別求出A組、B組、O組、H組的放射生物學參數(shù)。 【研究結果】 1.攜帶CK13基因的CK13-A、CK13-B質(zhì)粒和空載體質(zhì)粒pEGFP-N1滿足實驗要求。 2.以Lipofectamine~(TM)2000介導,CK13-A、CK13-B質(zhì)粒和空載體質(zhì)粒(pEGFP-N1)成功地轉(zhuǎn)染到HNE1中,轉(zhuǎn)染細胞組分別命名為A組、B組、O組和未轉(zhuǎn)染的HNE1細胞H組。 3.克隆形成實驗測定細胞劑量存活分數(shù),采用線性二次函數(shù)模型(LQ模型)和單擊多靶模型擬合細胞輻射劑量存活曲線。A組、B組、O組、H組在單擊多靶模型中D_0值分別是2.384、2.818、3.259、3.546;Dq值0.878、1.135、1.399、1.581;SF2值0.627、0.637、0.640、0.725。A組、B組、O組、H組在線性二次函數(shù)中α值分別是0.248、0.261、0.171、0.160;β值0.034、0.025、0.030、0.043;SF2值0.648、0.657、0.769、0.710;α/β值7.29、10.44、5.70、3.72。與HNE1細胞、轉(zhuǎn)染空載體質(zhì)粒O組的細胞比較,轉(zhuǎn)染CK13基因后,A組和B組的D_0、Dq、SF2均減小,α值、α/β值增大,證明CK13基因?qū)Ρ茄拾〩NE1細胞株有放射增敏性。 【結論】 CK13基因能明顯提高鼻咽癌HNE1細胞株的放射敏感性。
[Abstract]:[objective] to investigate the effect of CK13 gene on radiosensitivity of nasopharyngeal carcinoma (NPC) HNE1 cell line. The eukaryotic expression plasmid CK13-A carrying CK13 gene and CK13-B, empty plasmid pEGFP-N1 were transfected into nasopharyngeal carcinoma cell line HNE1 by liposome mediated transfection. Cell survival fraction was measured by clone formation assay. The cell survival curve was fitted by click multi-target model and linear quadratic function model, and the radiobiological parameters D0, DQ, N and 偽, 尾, 偽 / 尾, Sf2 were obtained to explore the relationship between CK13 gene and radiosensitivity of nasopharyngeal carcinoma HNE1 cell line. CK13 gene as target can provide a new idea for clinical improvement of radiotherapy sensitivity of nasopharyngeal carcinoma (NPC). [methods of study] 1. Objective Gene preparation; Recombinant DNA; transfection; screening. two銆，

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