脂质体介导CK13基因对鼻咽癌HNE1细胞株放疗敏感性的影响
发布时间：2019-06-12 09:17
【摘要】： 【研究目的】 本文探讨CK13基因对鼻咽癌HNE1细胞株放射敏感性的影响。我们采用脂质体介导转染方法将携带CK13基因的真核表达质粒CK13-A和CK13-B,空载体质粒pEGFP-N1分别转染到鼻咽癌细胞株HNE1中。克隆形成实验测定细胞存活分数。采用单击多靶模型和线性二次函数模型拟合细胞存活曲线,求出放射生物学参数D0、Dq、N和α、β、α/β、SF2值,探讨CK13基因与鼻咽癌HNE1细胞株放射敏感性的关系,以CK13基因为靶点进行相关研究,可为临床提高鼻咽癌放疗敏感性提供新的思路。 【研究方法】 1.目的基因的制备;重组DNA;转染;筛选。 2.携带CK13基因的质粒CK13-A、CK13-B和空载体质粒(pEGFE-N1)的转化、扩增、酶切与测序鉴定分析。 3.阳离子脂质体Lipofectamine~(TM)2000介导,将CK13-A;CK13-B质粒和空载体质粒pEGFP-N1转染至HNE1细胞中。分别命名为A组、B组、O组和H组。 4.克隆形成实验测定细胞剂量存活分数,采用线性二次函数模型(L-Q模型)和单击多靶模型拟合细胞辐射剂量存活曲线,分别求出A组、B组、O组、H组的放射生物学参数。 【研究结果】 1.携带CK13基因的CK13-A、CK13-B质粒和空载体质粒pEGFP-N1满足实验要求。 2.以Lipofectamine~(TM)2000介导,CK13-A、CK13-B质粒和空载体质粒(pEGFP-N1)成功地转染到HNE1中,转染细胞组分别命名为A组、B组、O组和未转染的HNE1细胞H组。 3.克隆形成实验测定细胞剂量存活分数,采用线性二次函数模型(LQ模型)和单击多靶模型拟合细胞辐射剂量存活曲线。A组、B组、O组、H组在单击多靶模型中D_0值分别是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组、B组、O组、H组在线性二次函数中α值分别是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。与HNE1细胞、转染空载体质粒O组的细胞比较,转染CK13基因后,A组和B组的D_0、Dq、SF2均减小,α值、α/β值增大,证明CK13基因对鼻咽癌HNE1细胞株有放射增敏性。 【结论】 CK13基因能明显提高鼻咽癌HNE1细胞株的放射敏感性。
[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 伪, 尾, 伪 / 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，
本文编号：2497904
[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 伪, 尾, 伪 / 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，
本文编号：2497904
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