| Part1Antitumor effects and mechanism of albendazole on pancreatic cancerObjective:To explore the impact of albendazole(ABZ)on human pancreatic cancer cell proliferation,migration and apoptosis in vitro,and to further verify the antitumor effects of ABZ on pancreatic cancer in vivo by establishing ectopic tumor model.Methods:Pancreatic cancer cells were treated with appropriate concentration of ABZ and gemcitabine(GEM,200 nM)for 12 or 48 hours.The MTT assay was used to test proliferation of ABZ and GEM to cultured pancreatic cancer.The wound healing assay and Transwell assay were used to test migration of ABZ and GEM to cultured pancreatic cancer.Annexin V-FITC Apoptosis Kit was used to test apotosis of ABZ and GEM to cultured pancreatic cancer.Heterotopic tumor model was established using SW1990 human PC cells to evaluate the anti-tumor effect of ABZ in vivo.Results:Compared with blank control group,the proliferation of human PC SW1990 and PANC-1 cell lines in ABZ treatment group was significantly inhibited,and the inhibition effect was concentration dependent.Exposure to GEM but not ABZ at the same drug concentration induces a better inhibition of pancreatic cancer cell proliferation,and the difference between these two groups showed statistically significance[(SW1990:Control vs.200 nM ABZ:96.12±3.27%vs.83.76±1.79%,P<0.01,200 nM ABZ vs.200 nM GEM:83.76v1.79%vs.74.79±2.81%,P<0.01);(PANC-1:Control vs.2 00 nM ABZ:78.67±1.55%vs.70.33±0.85%,P<0.01;200 nM ABZ vs.200 nM GEM:70.33±0.85%vs.66.17±1.25%,P<0.001)].The proliferation evaluation of SW1990 and PANC-1 cell lines by MTT assay is consistent with the results of cell colony formation assay.Compared with blank control group,the migration of human pancreatic cancer SW1990 and PANC-1 cell lines in ABZ treatment group was significantly inhibited,and the inhibition effect was concentration dependent.Exposure to GEM but not ABZ at the same drug concentration induces a better inhibition of pancreatic cancer cell migration,and the difference between these two groups showed statistically significance[(SW1990:Control vs.200 nM ABZ:317.33±23.33 vs.202.33±6.55,P<0.01;200 nM ABZ vs.200 nM GEM:202.33±6.55 vs.204.33±15.76,P<0.01);(PANC-1:Control vs.200 nM ABZ:382.33±4.99 vs 265.33±29.78,P<0.01;200nM ABZ vs.200nM GEM:265.33±29.78 vs.129.33±33.81;P<0.01)].The results of Wound healing assay were consistent with Transwell assay,which further verified the inhibitory effect of ABZ on the migration of human pancreatic cancer SW1990 and PANC-1 cell lines.Compared with blank control group,ABZ can significantly increase the apoptosis of pancreatic cancer cells,and the effect of ABZ on the induction of cell apoptosis was concentration dependent,and the difference between these two groups showed statistically significance(SW1990:Control vs.400 nM ABZ:14.45±0.78 vs.22.42±0.90,P<0.001;PANC-1:Control vs.400 nM ABZ:18.36±2.25 vs.28.16±4.18,P<0.05).SW1990 human pancreatic cancer carcinoma xenografts mice results showed that compared with control group,the mice treated with ABZ had a smaller tumor volume.The results of immunohistochemistry also showed that the expression level of proliferating antigen in ABZ treated mice was lower than that in the control group.Conclusions:ABZ can effectively inhibit pancreatic cancer cells proliferation and migration.And the possible mechanism,underlying these phenomenons,may be related to cell apoptosis induced by ABZ.Under the same concentration,gemcitabine(GEM)had a slightly better anti pancreatic cancer effect than ABZ.Part2The radiosensitivity effect and mechanism of albendazole on pancreatic cancerObjective:To evaluate the potential mechanism of albendazole(ABZ)in improving the radiotherapy resistance of human pancreatic cancer cell lines in vitro under hypoxia condition,and to further verify the radiosensitization effects and mechanism of ABZ on pancreatic cancer in vivo by establishing heterotopic tumor model.Methods:Pancreatic cancer cells were induced with hypoxia by 50?M COC12 and treated with vehicle control or ABZ(200 nM),followed by different doses of X-ray and incubator culture for 24 or 48 hours.The MTT assay and cell colony formation assay were used to test pancreatic cancer cell proliferation.Annexin V-FITC/PI double staining was performed to detect the apoptosis of pancreatic cancer cells.Western blot was used for detecting the expression level of hypoxia inducible factor-1?(HIF-1?)and basic fibroblast growth factor(bFGF)protein in pancreatic cancer cells.Heterotopic tumor model was established using PATU8988 human PC cells to evaluate the effect of ABZ on the radiosensitivity of pancreatic tumor,A total 24 mice were divided into 4 groups including:control,ABZ(Albendazole);Rad(Radiation);and ABZ+Rad(Albendazole+Radiation).TUNEL assay was used for detecting the apoptosis of human pancreatic cancer.And Ki-67 immunohistochemical staining was used to test pancreatic cancer proliferation.Results:At the same radiation dose,pancreatic cancer PATU8988 and SW1990 cells in hypoxia exhibit more rapid proliferation than that in normoxia However,ABZ can rectify the proliferation difference between these two groups.The proliferation evaluation of PATU8988 and SW1990 cell lines by cell colony formation assay is consistent with the results of MTT assay.The apoptosis of human pancreatic cancer cell lines PATU8988 and SW1990 in hypoxia group was more obvious than those cells in normoxia group,whereas ABZ can increase cell apoptosis of cells from hypoxia group.Western blotting assay showed that the expression level of HIF-1? protein and bFGF was significantly increased in the hypoxia group,while ABZ could decrease the expression level of those proteins.PATU8988 human pancreatic cancer carcinoma xenografts mice results showed that compared with the other three groups,the mice treated with X-ray and ABZ had the smallest tumor volume,the lowest expression level of proliferating antigen,the most obvious degree of apoptosis,and the lowest expression level of HIF-1? and bFGF protein.Conclusion:ABZ can increase sensitivity to radiotherapy in pancreatic cancer.And the possible mechanism underlying this phenomenon may be related to that ABZ can decrease the expression level of HIF-1? protein and bFGF. |
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