The Effect Of Rapamycin On Human Cholangiocarcinoma QBC939Cells And Mtor Expression

Objective:Cholangiocarcinoma QBC939cell lines was treated with themTOR inhibitor rapamycin,in order to explore the effect of rapamycin on thecholangiocarcinoma QBC939cells in vitro anti-proliferative and cell cycle,and toexplore its molecular mechanism.Methods:（1）Cell growth and proliferation of the QBC939cell lines inresponse to rapamycin was assessed by MTT assay;（2）Analyses and cell cycle of the QBC939cell lines in response to rapamycinwas assessed by flow cytometry;（3）The protein’s quantity of change of mTOR and P-mTOR were detectedusing Western blot pre-and post-treatment of rapamycin;（4）The mRNA levels of mTOR were detected using Realtime-PCR pre-andpost-treatment of rapamycin.Result:（1）The result of MTT technology indicated that the group withrapamycin concentration in2.5,5,10,20,40,80nM can reduce the proliferation rate ofQBC939; compare the group with rapamycin in all concentration with the controlgroup have significant differernce (P <0.01);DMSO group compared with controlgroup without statistical significance(P>0.05).（2）The result of FCM technology indicated that the group with rapamycinconcentration in2.5,5,10,20,40,80nM can increase the apoptosis rate of QBC939;compare the group with rapamycin in all concentration with the control group havesignificant differernce (P <0.01); DMSO group compared with control group withoutstatistical significance(P>0.05);compare the group of concentration in2.5nM with thecontrol group,treatment of rapamycin was shown to increase in the proportion of cellin the G1phase and decrease in the proportion of cell in the S phase, the result have significant differernce (P <0.01).（3）The result of Western blot indicated that compared the protein’s quantity ofchange of mTOR in group with rapamycin concentration in5,10,20nM with thecontrol group have no significant difference. compared with the control group theprotein’s quantity of change of P-mTOR in group with rapamycin in all concentrationwas shown to reduce,the result of5nM have difference(P<0.05), the result of10,20nMhave significant difference(P <0.01).（4）The result of Realtime-PCR indicated that compared the mRNA’s quantityof change of mTOR in group with rapamycin concentration in5,10,20nM with thecontrol group have no significant difference(P>0.05).Conclusion：（1）Rapamycin can inhibit growth and proliferation of theQBC939cell lines.（2）Rapamycin can induce apoptosis of the QBC939cell lines and inhibit thecell cycle.（3）Rapamycin can not change the protein’s and mRNA’s quantity of mTOR（4）Rapamycin can inhibit and lower the protein levels of p-mTOR of QBC939...