Effect Of Lentivirus Mediated XAF1 Gene Overrxpression Of The Radiosensitivity Of Radioresistant Human Nasopharyngeal Carcinoma Cell Line CNE-2R

Objectives: To discuss the influence of tumor suppressor gene XAF1 to the growth,proliferation,apoptosis,and radiosensitivity of nasopharyngeal carcinoma,and provide theoretical evidence for new targeted gene therapy of nasopharyngeal carcinoma.Methods:In this study,cultured human nasopharyngeal carcinoma cell line CNE-2R,which were transfected XAF1 by lentivirus in vitro.Proliferation and apoptosis of CNE-2R cells after transfection was observed by using CCK-8 and Federation of Canadian Municipalities(FCM)method.RT-PCR was used to detect the expression levels of XAF1 mRNA of CNE-2R cell before or after infection,and Western Blot was used to detect the protein expression levels.In addition,in this study,Infection the cells that before or after Infection were irradiated.Use FCM and Colony formation assay to explore the impact of XAF1 to the CNE-2R radio sensitivity.Results:(1)After the XAF1 infection of CNE-2R cells.CNE-2R cells transfected with XAF1 were taken as experimental group(XAF1-CNE-2R),those transfected with control virus and without any treatment were taken ascontrol group(NC-CNE-2R)and blank group(CNE-2R).(2)By qRT-PCR and Western Blot,we found that the expression of XAF1 was higher in experimental group,and lower in control group and blank group.(3)By CCK-8 assay,we found that experimental group showed a low cell survival rate compared with control group and blank group.No significant difference between the control group and blank group.(4)By Colony formation assay,we found Radio sensitivity of experimental group cells was significantly higher,while the control group and blank group had no significant effect.(5)By flow cytometry(FCM)to detect the cell cycle,we found that XAF1 can make the experimental group cell cycle arrest in G2 / M compared with control group and blank group.(6)By flow cytometry(FCM)to detect the cell apoptosis,we found,in experimental group,XAF1 combination with radiation can greatly increase the rate of apoptosis.Conclusions:(1)Lentivirus may well transfected the human nasopharyngeal carcinoma cell line CNE-2R.We build the XAF1-CNE-2R which overexpression the XAF1.(2)XAF1 can increase the radiation sensitivity of human nasopharyngeal carcinoma cell line CNE-2R.XAF1 is expected to become the ideal target genes for gene therapy of nasopharyngeal carcinoma.