Radio-sensitivity Effect Of CK13 Gene Mediated By Lipofectin On Nasopharyngeal Carcinoma Cell Strain HNE1

[Purpose]This article discussed the radiosensitive effect of the CK13 gene on nasopharyngeal carcinoma cell strain HNE1.We used Liposome mediated let CK13-A,CK13-B plasmid and empty plasmid pEGFP-N1 transfected cells HNE1.We measured survival fraction by colony forming assay;then fit cell survival curve by using multi-target single-hit mode and linear quadratic model;in turn,calculate radiobiological parameters of D0,Dq,N,andα,β,α/β,SF2.Using CK13 as a target, this study investigated the relationship between CK13 and NPC radiosensitivity.To privide a new idea for clinical nasopharyngeal carcinoma radiotherapy.[Materials and methods]1.preparation of target gene;Recombinant DNA;Transfection;Screening.2.Transformation,amplification,restriction enzyme digestion and sequencing of the plasmids,including CK13-A,CK13-B and empty vector plasmid(pEGFP-N1).3.Transfected HNE1 cells with CK13-A,CK13-B and empty plasmid pEGFP-N1 mediated by Lipofectamine^TM2000.The transfected cells were named as group A,B,O and H respectively.4.The colony forming assay was performed to determine cells under different doses of irradiation survival fraction.Using linear quadratic model and multi-target single-hit mode fit cell survival curve,calculated radiobiological parameters of A, B group,O group,H group.[Results] 1.The CK13-A,CK13-B plasmid which carrying CK13 gene and pEGFP-N1 plasmid fit our study requirement.2.CK13-A,CK13-B and pEGFP-N1 plasmid were transfected into the nasopharyngeal carcinoma cell strain HNE1 successfully.The transfected cells were named as group A,B and O respectively,none transfected HNE1 cells named as H.3.The colony forming assay was performed to determine cells under different doses of irradiation survival fraction.Using the single-hit multi-target model and the linear-quadratic model to the fit surviving fraction after irradiation,The A group, B group,O group and H group cells had the D₀ parameters of 2.384,2.818,3.259, 3.546 respectively;the Dq parameters of 0.878,1.135,1.399,1.581 respectively, the SF2 parameters of 0.627,0.637,0.640,0.725 respectively.In the single-hit multi-target;Theαparameters of 0.248,0.261,0.171,0.160 respectively;theβparameters of 0.034,0.025,0.030,0.043,respectively;the SF2 parameters of 0.648,0.657,0.769,0.710 respectively;α/βparameters of 7.29,10.44,5.70, 3.72 respectively,the SF2 parameters of 7.29,10.44,5.70,3.72 respectively.In the linear-quadratic model respectively.Compared with HNE1 cells with pEGFP-N1 D0,Dq,SF2 decreased andα,α/βincreased were seen in CK13 transfected cells after radiation.Prove CK13 gene enhanced the radio-sensitivity of nasopharyngeal carcinoma cell strain HNE1.[Conclusions]CK13 gene could enhance the radiosensitivity of nasopharyngeal carcinoma cell strain HNE1.