The Impact Of Tumor Suppressor Gene XAF1to Lung Adenocarcinoma

Objectives: The purpose of the study was to investigate the impact of tumorsuppressor gene XAF1to the Growth, proliferation, apoptosis, and radiosensitivity of lungadenocarcinoma, and provide theoretical evidence for new targeted gene therapy of lungadenocarcinoma.Methods: In this study, cultured A549lung adenocarcinoma cell line, A549cell lineswere transfected by recombinant adenovirus Ad5/F35-XAF1in vitro. proliferation andapoptosis of A549cells after transfection was observed by using MTT and Federation ofCanadian Municipalities (FCM) method. RT-PCR was used to detect the expression levelsof XAF1and DR4mRNA of A549cell before or after infection, and, Western Blot wasused to detect the protein expression levels. in addition, in this study, Infection the cellsthat before or after Infection were irradiated. Use FCM and Colony formation assay toexplore the impact of XAF1to the A549radiosensitivity.Results:(1) After the adenovirus Ad5/F35-XAF1infection of A549cells, cell deathincreases with the increase of the concentration.(2) By RT-PCR and Western Blot, wefound that the expression of XAF1and DR4were in a dose-dependent increased in A549infected cells, and were higher than those PBS and Ad5/F35-Null treated cells in theexpression.(3) By MTT assay, we found The XAF1showed a dose-and time-dependentinhibition of the proliferation of A549cells.(4) Cell apoptosis were detected by flowcytometry (FCM), XAF1was a dose-and time-dependent induction of apoptosis of A549cells.(5) After the treatment by Ad5/F35-XAF1and Irradiation, and detected by FCM, wefound, in A549cells, XAF1combination with radiation can greatly increase the rate ofapoptosis.(6) By Colony formation assay, we found Radiosensitivity of A549cells wassignificantly higher after infected by Ad5/F35-XAF1,while the PBS group and the Ad5f35-Null group had no significant effect. Conclusion:(1) The adenovirus Ad5/F35-XAF1and the report virus Ad5/F35-EGFP are strong inthe infection of A549cells, and A549cells can be successfully transiently transfected, thevirus is a very good gene vector.(2) The expression or even over-expression of XAF1can increase the apoptosis rateof A549cells, XAF1is expected to be an ideal target genes in the treatment of lung cancer.(3) In A549cells，the expression of DR4is increases along with the increase of theexpression of XAF1, So the combination XAF1and TRAIL may be a better treatment oflung adenocarcinoma, and even, XAF1may play a role in the reversal of TRAILresistance.(4) The A549cells in which XAF1is higher expression, the radiosensitivity is alsoincreased, XAF1might become a Radiation sensitizer of Lung adenocarcinoma.