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Objective To investigate the inhibitory effects and mechanisms of a nature product derivate oridonin on in?vasion of human lung cancer. Methods Human lung cancer A549 and PC-9 cell lines were treated with oridonin. MTS as?say was used to determine cell proliferation. Transwell assay was used to determine the cell invasion, and adhesion assay to determine the cell adhesion. Flow cytometry was used to determine cell cycle. Western blotting and realtime-PCR were used to detect expression levels of CDK1, mTOR, p53, p21, E-cadherin, CD44,β-catenin, uPA, MMP-2/9, p-AKT and p-Src. The luciferase reporter assay was used to detect the NF-κB promoter activity. Results In vitro proliferation, invasion and adhesion of A549 and PC-9 cells were significantly inhibited by oridonin. The cell cycle was halted by G2/M phase, and ex?pressions of E-cadherin, p53 and p21 were promoted, while expressions of CDK1, mTOR, CD44,β-catenin, uPA, MMP-2/9, p-AKT and p-Src and promoter activity of NF-κB were down-regulated. Conclusion Oridonin is able to inhibit the in vitro invasion of human lung cancer A549 and PC-9 cell lines, which might be correlated with its abilities to regulate the ty?rosine kinase activity.
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Objective To investigate the inhibitory effects of oridonin combined with gemcitabine on pancreatic cancer SW1990 cells in vitro, and the potential mechanisms thereof. Methods The pancreatic cancer SW1990 cells were treated with vehicle alone and various concentrations (10,20,40,80 and160μmol/L) of oridonin, followed by 24, 48 and 72 h cell culture. Effects of oridonin on cell proliferation were determined by using a CCK-8 kit. SW1990 cells were treated with oridonin (40μmol/L) and gemcitabine (20μmol/L) alone or together for 48 h, and the untreated cells were used as the con-trol. The cell survival rate was detected by CCK-8 assay. Apoptosis induction was assessed by using Annexin V-FITC kit. Semi-quantitative RT-PCR was used to examine the changes of NF-κB mRNA and XIAP mRNA expressions. Results Oridonin inhibited the growth of pancreatic cancer SW1990 cells in a dose-and time-dependent manner. Compared with the other groups, the cell survival rate was significantly lower in the combination group (P<0.05). Oridonin combined with gemcitabine induced a higher percentage of apoptosis in pancreatic cancer cells than that of oridonin or gemcitabine alone (P<0.05). Moreover, the expressions of NF-κB and XIAP mRNA in pancreatic carcinoma cells were obviously down-regu-lated in combination group (P<0.05). Conclusion Oridonin can enhance the antitumor effect of gemcitabine on pancreatic cancer in vitro, which may be related to through the down-regulation of NF-κB and its downstream of XIAP, and then induc-ing cell apoptosis in pancreatic cancer.
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Objective To study the apoptosis-inducing effect of Oridonin on PC-3 cells line and the role of Survivin in the process.Methods After PC-3 cells were incubated with different concentrations of Oridonin,cell viability was analyzed with MTT assay.The percentage of earlier apoptosis cell was analyzed by flow cytometry.The protein expression of Survivin in PC-3 cells were detected by Western blot and fluorescent quantitative PCR.Results Oridonin effectively inhibited the proliferation of PC-3 cells in a concentration-time dependent way.After PC-3 cells were treated with Oridonin ( 2.5,5,10,20,40 μmol/L)for 48 hours,the cytotoxicity index were 9.2％,25.3％,39.3％,77.2％,92.5％ and the IC50 of PC-3 cells was 10.29 μmol/L,respectively.Flow cytometry was used to detect the effect of different concentration of Oridonin (0,10,20,40 μmol/L) for 48 hours,the apoptotic rates of PC-3 cells were 4.8％,15.4％,19.5％,27.4％ ( P ＜ 0.05).Oridonin down-regulated Survivin protein in a concentration-dependent way in PC-3 cells.Conclusions Oridonin can induce the apoptosis of PC-3 cells by a concentration-dependent manner.Oridonin can induce the apoptosis of PC-3 cells by down-regulated Survivin protein.
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Objective To investigate the apoptosis-inducing effect of oridonin combined with valproic acid( VPA)on leukemic cell line HL-60,and study the feasibility of oridonin combined with VPA to be used in clinical practice. Methods Oridonin of 6-12 μmol/L combined with VPA of 0. 5-1 mmol/L were added in exponential growth HL-60 cells respectively. Cell count assays were used to measure the growth inhibitory effect of oridonin combined with VPA or alone. Flow cytometry was used to evaluate apoptosis with Annexin V and propidium iodide (PI) double staining. Results Combined use of oridonin and VPA could synergistically inactivate HL-60 cells,and inhibit the cell proliferation and induce apoptosis in a dose-and time-dependent manner (P < 0.05 ). Conclusion Oridonin has a synergistic effect combined with VPA. Oridonin has a promising prospect in clinical use of leukemia.
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The success of ATRA and ATO in APL treatment furuishes the first model of molecular target-based induction of differentiation and apoptosis.Two drugs all target PML-RARα oncoprotein through different moieties and induce APL cells differentiation and apoptosis.In acute myeloid leukemia M2b,we reveal that gain-of-function of C-kit coexists with persistent AML1-ETO,suggesting that abnormal C-kit may serve as a therapeutic target in AML L-M2b Oridonin becomes a potential candidate target drug which specifically degrades AML1-ETO protein. Experiment in vitro proves the combinatorial effectiveness of imatinib and cytarabin in the treatment of chronic myelogenous leukemia(CML).Imatinib is an inhibitor of abnormal protein tyrosine kinases activity. While arsenic agent triggers the degradation of bcr-abl, and combination of the two drugs in treating CML leads to a better outcome.
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AIM To investigate the inhibition and apoptosis mechanism of GBC-SD cells induced by rubescensine B. METHODS Using MTT, convert microscopy, electron microscopy, flow cytometry, an immunohistochemical assay, and spectrofluorometry demonstrate the presence and pathogenesis of apoptosis after treated by rubescensine B. RESULTS After exposure to Rubecensine B GBC-SD cells were induced to apoptosis in dose-dependent manner, and the level of Bcl-2,p53,C-myc,Fas/APO-1 were decreased within 24 hours, reversely the activity of Caspase-3 was enhanced with the appearance of apoptosis. CONCLUSION Rubecensine B can induce GBC-SD cells apoptosis related to Bcl-2,p53,Fas/APO-1 and C-myc.