Function Of E545K Mutation In PIK3CA In Gallbladder Cancer

Part ? Clinicopathological and Prognostic Implications of PIK3 CA E545K Mutation in Patients with GBCObjective To investigate the clinical significance of PIK3 CA E545K mutation in the GBC.Methods The GBC tissue specimens were obtained from 130 patients who underwent cholecystectomies(without prior radiotherapy or chemotherapy)and the PIK3 CA sequences were obtained by targeted sequencing.Moreover,the correlation between PIK3 CA E545K mutation in GBC and clinicopathologic parameters was analyzed.The Kaplan-Meier test was used to measure the survival of GBC patients.Results The rate of PIK3 CA E545K mutation in GBC patients was 6.15%.There was no significant correlation between E545 K mutation and clinicopathologic parameters of GBC patients,but the survival of GBC patients was correlated with E545 K mutation significantly(P<0.05).Conclusions There was no significant correlation between E545 K mutation and clinicopathologic parameters of GBC patients.The possible cause was that the sample size was not sufficiently large to reveal such relationships.The survival of GBC patients was correlated with E545 K mutation significantly and the prognosis of patients with E545 K mutation was worse than patients without this mutation.Part ?The Function and Mechanism of PIK3 CA E545K Mutation in GBC Cell LinesObjective To investigate the function and mechanism of PIK3 CA E545K mutation in the GBC cell lines.Methods The proliferation,migration,invasion and clone formation of PIK3 CA knockdown,WT PIK3 CA overexpression,E545 K PIK3CA overexpression and normal cells were tested by CCK8 assay,Transwell assay and clone formation assay in vitro.The mechanism of E545 K was analyzed by Western Blot and Co-IP assay in vitro.The function of E545 K was tested by subcutaneous tumor of nude mice in vivo.Results The E545 K mutation promoted proliferation,migration and invasion of GBC cells significantly.The E545 K mutation of PIK3 CA promotes gallbladder carcinoma progression through enhanced binding to EGFR and activating downstream akt activity.E545 K promoted tumor proliferation in mice.Conclusions The E545 K mutation of PIK3 CA promoteed gallbladder carcinoma progression through enhanced binding to EGFR.Part ? The Function and Mechanism of PIK3 CA E545K Mutation in GBC Cell LinesObjective To investigate the function of PI3 K selective inhibitor on GBC cell lines with PIK3 CA E545K mutation.Methods The proliferation of WT PIK3 CA overexpression and E545 K PIK3CA overexpression cells under treatment with PI3 K p110? selective inhibitor,A66,was tested by CCK8 assay in vitro.The mechanism of A66 was analyzed by Western Blot and Co-IP assay in vitro.The function of A66 was tested by subcutaneous tumor of nude mice in vivo.Results PI3 K selective inhibitor,A66,suppressed proliferation of WT PIK3 CA overexpression and E545 K PIK3CA overexpression GBC cells significantly.A66 functioned without depressing the expression of PI3 K p110?.A66 suppressed tumor proliferation in mice.Conclusions The PI3 K selective inhibitor,A66,suppressed gallbladder carcinoma proliferation.And this effect may function through converting the PI3 K p110? to inactived form.