| Part ⅠRole of MMI-166in proliferation and apoptosis of SW1990pancreatic cancer cellsObjectiveTo investigate the proliferation and apoptosis effect of MMI-166on human pancreatic cancer cell line SW1990cells.MethodsMMI-166in different concentrations (25μg/ml/50μg/ml/100μg/ml) were added to cultures of human pancreatic cancer cell line SW1990. Effects of MMI-166on cell proliferation were detected by3-(4,5-dimethyl-2-thiazole)-2-5-biphenly-tetrazole bromide (MTT) method and effects on cell apoptosis were tested by fluorescence microscope and flow cytometry (FCM) at24and48h after treatment.ResultsAfter sw1990cells were treated by MMI-166at different concentrations, MTT showed that MMI-166have significant inhibition on human pancreatic cell lines in a time-and concentration-dependent manner. Under the fluorescence microscope, the early stage of apoptosis cells were stained by Annexin V (green fluorescence); Late apoptotic cells and necrotic cells were stained by PI(red fluorescence). FCM showed that cells apoptosis rate increased along with the increase in the drug’s concentrations.Conclusions MMI-166has proliferation inhibition and apoptosis-inducing effect on human pancreatic cell lines in a time-and concentration-dependent manner. Part IIRole of MMI-166in nude mouse xenografts of human pancreatic cancerObjectiveTo investigate of the MMI-166on the expression of MMP-2, MMP-9and the cell apoptosis of nude mouse xenografts of SW1990human pancreatic cancer cells.MethodsEstablishment of control and experimental groups, randomly, the human pancreatic cancer xenograft model of SW1990was constructed. The control group was treated with normal saline, and experimental group was treated with MI-166(200mg/kg/day). The tumor volume and tumor inhibition rate was measured by vernier caliper through length and short diameter. The expression of MMP-2and MMP-9protein was observed using immunohistochemistry in the tumor tissues. Apoptosis index (AI) was detected by deoxynucleotidyl transferase-mediated nick end labeling (TUNEL method).ResultsThe tumor volume of MMI-166group (1252.30±464.84mm3) was less than the control one (2241.82±208.06mm3), significantly. The inhibition rate was34.47%between experimental groups(treat with MMI-166)(1.42±0.146g) and control group (2.17±0.197g). The expression of MMP-2(2.8±1.095%) and MMP-9(2.6±1.517%) protein was significantly downregulated in MMI-166group, compared with the control group. AI in the experimental group (75.6±9.710)%was higher than the control group (17.4±10.139)%, significantly.ConclusionsThe mechanism of MMI-166inhibiting pancreatic tumor growth and inducing apoptosis may be related to the suppression of MMP-2and MMP-9protein expression. Part ⅢRole of MMI-166in expression of apoptosis-related proteins of pancreatic cancer SW1990cellsObjectiveTo investigate the mechanism of MMI-166to promote apoptosis in human pancreatic cancer cell line SW1990.MethodsEstablishment of control and experimental groups, randomly, the human pancreatic cancer xenograft model of SW1990was constructed. The control group was treated with normal saline, and experimental group was treated with MMI-166(200mg/kg/day). Apoptosis index (AI) was detected by deoxynucleotidyl transferase-mediated nick end labeling(TUNELmethod).The expression of p53, c-myc, bax, bcl-2, survivin, caspase-1and fas proteins,which are related with apoptosis, were observed using immunohistochemistry in the tu-mor tissues,then to find the protein whose expression is definitely different between the two groups. MMI-166of different concentrations (0,50,100μg/ml) were used to treat hunman pancreatic cancer SW1990cell for24h. Then the proteins expression were measured by western blot.Results1. AI in the control group (21.3±2.214) was lower than the experimental group (81.1±7.852), significantly. The IOD value of c-myc (7714.516±2228.915) and survivin (4594.058±1239.530) proteins of MMI-166group was significantly different from the control group, with a significant difference(P<0.01). Twenty-four hours after MMI-166treatment of different concentrations (0,50,100μg/ml), the c-myc expression was (0.169±0.007),(0.098±0.003)and(0.073±0.008), with a significant difference(P<0.05); the survivin expression was (0.391±0.006),(0.395±0.006),(0.382±0.004),without a significant difference(P=0.069).ConclusionsMMI-166can induce cell apoptosis in vitro of SW1990by down-regulating the expression of c-myc. |
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