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Objective To evaluate the effect of ANO9 on the radiosensitivity of pancreatic cancer cell AsPC-1,aiming to provide new targets for clinical radiotherapy of pancreatic cancer.Methods Western blot was performed to detect the expression of ANO9 in pancreatic cancer cell lines (BxPC-3,PANC-1,AsPC-1)and normal pancreatic cell line (HPNE).The AsPC-1 cell line with stable silencing ANO9 was constructed by using lentivirus and validated by Western blot.MTT assay was adopted to detect the cell viability of AsPC-1 with stable silencing ANO9 after irradiation.Colony formation assay was conducted to evaluate the effect of silencing ANO9 upon the radiosensitivity of AsPC-1 cells.Western blot was performed to assess the effect of ANO9 silencing on the expression of EGFR/ERK signaling protein.Results The expression levels of ANO9 were significantly up-regulated in three pancreatic cancer cell lines compared with that in the normal pancreatic cell line HPNE (t =7.426,5.543,11.850,all P＜0.05).After silencing ANO9,the expression level of ANO9 protein was significantly down-regulated than that in the control group (t =9.670,P＜0.05).The AsPC-1 cells with stable silencing ANO9 were successfully constructed.The sensitivity of AsPC-1 cells to irradiation was significantly increased after silencing ANO9,and the sensitivity enhancement ratio was 1.566.The expression levels of EGFR/ERK signaling proteins (EGFR and p-ERK 1/2) were significantly downregulated after silencing ANO9 (t =7.949,13.160,both P＜ 0.05).Conclusions Silencing ANO9 can significantly increase the sensitivity of AsPC-1 cells to radiotherapy,which is probably associated with the inhibition of EGFR/ERK signaling transduction.ANO9 might be a new therapeutic target for preventing the progression of pancreatic cancer.
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Objective To observe the influence on the sensitivity of pancreatic cancer cell line BxPC-3 to gemcitabine of silencing PAUF gene.Methods BxPC-3 cells,which overexpress PAUF,was stably transfected with PAUF-shCtrl and PAUF-shRNA to establish BxPC-3_shCtrl and BxPC-3_shPAUF cells as control and experiment group.Then the mRNA and protein expression level of PAUF in these two cell lines were detected by RT-PCR and western blot,respectively.The growth inhibition rates of these two cell lines treated with different concentrations of gemcitabine (0,3.1,6.25,12.5,25,50,100,200 nmol/L) were detected by MTT.Apoptosis rates in the cells treated with different concentrations of gemcitabine (0,75,100 nmol/L) were then observed by flow cytometry.Results The relative PAUF mRNA expression level in BxPC-3_shCtrl and BxPC-3 cells were 1.00 ± 0.06 and 0.83 ± 0.07,which were significantly high er than that in BxPC-3_shPAUF cells (0.25 ± 0.02;both P ＜ 0.05).The relative PAUF protein expression level in BxPC-3_shCtrl and BxPC-3 cells were 0.89 ± 0.07 and 0.95 ± 0.04,which were significantly high er than that in BxPC-3_shPAUF cells (0.31 ± 0.03;both P ＜ 0.05).The IC50 value of gemcitabine to BxPC-3_shCtrl cell was (22.88 ± 2.43) nmol/L,which was significantly higher than that of BxPC-3_shPAUF cells [(1.06 ± 0.02) nmol/L;P ＜ 0.05];apoptosis rate of BxPC-3_shPAUF cells treated by gemcitabine increased faster than that of BxPC-3_shCtrl cells.Conclusion PAUF silencing could greatly enhance the sensitivity of BxPC-3 cells to gemcitabine.
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OBJECTIVE:To observe the inhibitory effect of phenolicalkaloids of menispermum dauricum(PAMD)on the human pancreatic cell line(BXPC-3)tumor and study the effect of PAMD on serum content of inducible nitric oxide synthase(iNOS)in tumor-bearing mice.METHODS:The BXPC-3 tumor-bearing mice model was established,and the model mice were randomly assigned to 6 groups:model control group,blank control group,cyclophosphamide group,3 PAMD groups(high,medium and low dosages).The blank control group was not inoculated with tumor strain but given same volume of normal saline.The tumor-inhibition ratio of PAMD was detected and the content of iNOS of in peripheral blood of tumor-bearing mice was determined by fluorospectrophotometry.RESULTS:PAMD(high,medium and low dosages)showed marked inhibitory effect on tumor growth of BXPC-3 cell line in tumor-bearing mice,with the inhibition ratios at 34.91%,52.83% and 41.51%,respectively.As compared with model control group,PAMD-treated groups showed significantly lower content of iNOS in peripheral blood of tumor-bearing mice.CONCLUSION:PAMD showed marked inhibitory effect on tumor growth of BXPC-3 cell line in tumor-bearing mice,which might be attributed to the lowered content of iNOS.
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Objective To identify effects of bile acids on pancreatic cancer, The ultrastructure and growth of PANC-1 and MIA PaCa-2 cell lines in crude bile modified medium were studied. Methods The growth of PANC-1 and MIA PaCa-2 cells in RPMI 1640 with or without 1％, 2％ and 4％ of the purified crude bile (containing total bile acids 10.17mmol/L) was assessed for 2, 4, 6, 8d by using MTT assay to determine inhibitory rate. The cell surface and intracellular ultrastructure of PANC-1 cells was investigated by SEM and TEM at 24h and 48h, respectively. Re sults The proliferation of both cell lines in bile treated medium were greatly retarded (P ＜0.001). The inhibitory rate of 1％, 2％ and 4％ bile on Panc-1 cells in 4d were 38％, 60％ and 66％, respectively (P ＜0. 05), on MIA PaCa-2 cells at 4d were 28％, 39％ and 52％, respectively (P ＜0. 05). The cells grown in bile for 48h lost their mi crovilli, their mitochondria and other organelles became vacuolated. Conclusion The bile acids in bile has cytotoxicity on PANC-1 and MIAPACA-2 cells, which may inhibit pancreatic cancer progress in patients clinically.