Effect of celastrol on proliferation and apoptosis of human esophageal squamous cell carcinoma KYSE180 cells and its mechanism
Objective To explore the effect of celastrol on the proliferation and apoptosis of human esophageal squamous cell carcinoma KYSE180 cells and the possible mechanism.Methods KYSE180 cells in logarithmic growth phase were added with 0.5,1 and 2 μmol/L of celastrol respectively,and the blank control group was set up.MTT assay and colony formation test were used to detect the proliferation of cells;the apoptosis of cells was assessed by flow cytometry;Transwell assay was used to measure the migration and invasion ability of cells;the mRNA transcription levels of P53,FAS and DR5 were determined by qRT-PCR;the expression levels of Caspase-3,Caspase-9,Bax,Bcl-2 and PARP proteins were detected by Western blot.Results Compared with the control group,cell proliferation in 2 μmol/L celastrol group was significantly inhibited(F=10.9,P<0.05),and the numbers of clones in 0.5 and 1 μmol/L celastrol groups decreased significantly(F=457.9,P<0.05);the numbers of cell invasion and migration decreased significantly in 1 and 2 μmol/L celastrol groups(F=751.7-1 134.0,each P<0.001);the apoptosis rates in 0.5,1 and 2 μmol/L celastrol groups increased signifi-cantly(F=243.0-728.0,each P<0.05);the mRNA transcription levels of P53,FAS and DR5 in 1 and 2 μmol/L celastrol groups significantly increased(F=71.0-539.3,each P<0.01);the protein expression levels of Caspase-3,Cas-pase-9 and Bax in 1 and 2 μmol/L celastrol groups significantly increased(F=142.1,35.3 and 347.6,respectively,each P<0.01),the level of PARP in 2 μmol/L celastrol group significantly increased(F=877.6,P<0.01),and the expression of Bcl-2 in 0.5,1 and 2 μmol/L celastrol groups decreased significantly(F=59.2,each P<0.01).Conclusion Celastrol can inhibit the proliferation and promote the apoptosis of KYSE180 cells,probably through the up-regulation of Caspase-3/9,Bax,PARP protein expression and the down-regulation of Bcl-2 protein expression.