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Objective To explore raddeanin A(RA) inhibiting the proliferation of HCT116 cells and its related mechanism.Methods The MTT method was used to observe the RA inhibition on the proliferation of gastric cancer cells HCT116.Flow cytometry was used to detect the RA effects on cell apoptosis.Transmission electron microscope(TEM) was used to observe RA induced autophagy.Western blot was used to test expression of apoptosis and autophagy related proteins.Results RA could significantly inhibit the proliferation of HCT16 cells with concentration and time dependence;double layer membrane of autophagosome was detected by TEM;FCM showed that RA induced apoptosis in HCT116 cells,moreover the apoptosis rate was increased with the concentration increase;Western blot showed that the expression of autophagy related proteins(Beclin1 and LC3) was increased,the expression of apoptosis inhibition protein Bcl-2 was decreased.On the contrary,the expression of apoptosis promotion proteins(Bax,Cleaved-caspase-3,Cleaved-PARP) was increased,the expression of mTOR protein in the mTOR signal pathway was increased,while the p-mTOR protein expression was decreased.When the mTOR pathway inhibitor rapamycin(RAPA) was added to cells,Beclin-1,LC3,Cleaved-caspase-3,Cleaved-PARP proteins were increased,and apoptosis rate was also increased.Conclusion RA can inhibit the proliferation of HCT116 cells and can induce cellular autophagy and apoptosis,its mechanism may be realized by regulating the mTOR signal pathway.
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Objective To investigate the effects of Raddeanin A(RA)on apoptosis and cell cycle of hu-man colon cancer HT29 cells,and discuss the possible mechanism.Methods Different concentration of RA 1,2, 4,8,16μmol/L disposed HT29 cells 12,24,48 hour,3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bro-mide(MTT)assay was used for the cell proliferation;Hoechst33258 staining was performed to observe the effects of RA on apoptosis morphology;Flow cytometry(FCM)was devoted to calculate apoptosis rate and detect cell cycle;The expression of Cleaved-caspase-3,Cleaved-PARP,CyclinB,CDK1 proteins and other proteins were tested by Western blot(WB). Results RA could inhibit the proliferation of HT29 cells,and associated with concentration and time(P<0.05);The IC50of 12 h,24 h,48 h were(9.31 ± 0.63)μmol/L,(4.88 ± 1.02)mol/L,(3.60 ± 0.89) μmol/L.The nucleus pycnosis,fracture were detected by Hochest33258 staining,RA induced apoptosis in HT29 cells;Western blot(WB)assay showed that the expression of Cleaved-caspase-3,Cleaved-PARP rised.HT29 cells were arrested at the period of G2/M,and the CyclinB,CDK1 proteins down-regulated.WB result showed that PI3K, AKT proteins up-regulated,while p-PI3K,p-AKT protein down-regulated.Conclusion RA can inhibit the prolifer-ation of colon cancer HT29 cells and its mechanism is probably by regulating the PI3K/AKT signaling pathways which through inducing apoptosis and blocking the cell cycle.
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0.05).(CONCLUSIONS) The HPV16,18 infection can induce incidence of breast cancer.The infection rates of HPV16,18 are not related to histological types,and lymph nodes metastases of breast cancer.
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Objective To study the role of Ki- 67 and anti- cytokeratin antibody 34?E12 in the differential diagnosis of prostate cancers. Methods 24 cases of prostate carcinoma and 30 cases of benign prostate hyperplasia were studied with immunohistochemical technique(SP). Results In 24 prostate cancers the average Ki- 67 index was 0.45?0.21, and that in 30 cases of benign hyperplasia was 0.12?0.09, the expression of Ki- 67 is higher in prostate carcinoma than benign prostate hyperplasia, there was static significance (P