An Experimental Study Of Resveratrol And Its Combination With Cisplatin In Inhibiting Breast Cancer Metastasis Through The EMT Pathway

Objective: To investigate the effect and mechanism of resveratrol and its combination with cisplatin in inhibiting breast cancer metastasis through the EMT approach.Methods:(1)In vivo experiments were conducted in nude mice with human breast cancer MDA-MB-231 cells xenograft tumor model to observe the changes of tumor weight and body weight of nude mice with resveratrol,cisplatin and combination of resveratrol and cisplatin.The changes of nodules of lung metastasis in nude mice were observed by HE staining in lung tissue sections.The changes of AST,ALT,Cr and BUN in serum were detected with the kit.Expression of metastasis related proteins in tumor tissues was observed by Westrn blot.(2)In vitro experiments,human breast cancer cell MDA-MB-231,MDA-MB-436,MDA-MB-453,BT-549 cellsand TGF-?1 induces EMT in MDA-MB-231 cells model were used.MTS assay was used to detect the effects of resveratrol and its combination with cisplatin on cells' viability.Cell scratch assay was used to detect the migration ability of MDA-MB-231 cells.Transwell assay was used to detect the effect of drugs on cells' migration and invasion ability.Western blot was used to detect the effect of drugs on the expression of EMT proteins and EMT-related pathway proteins.Results:(1)Compared with the model group,100 mg/kg resveratrol can significantly inhibit the growth and lung metastasis of human breast cancer MDA-MB-231 cells in nude mice,and the tumor weight and number of metastatic nodules have statistically significant(P < 0.05),and the is no statistical difference between nude mice in weight and the liver and kidney function(P > 0.0.5).The migration of MDA-MB-231 cells were significantly inhibited by resveratrol 12.5,25 and 50 ?M for 24 h,and the migration rates were 85%,73%,and 42%(P < 0.05).Resveratrol 50 ?M could reverse the mesenchymal morphological changes in TGF-?1-treated MDA-MB-231 cells,and inhibited TGF-?1-increased cell migration.Western blot and immunofluorescence experiments observed that resveratrol can reduce the expression of mesenchymal marker protein Vimentin and improve the expression of epithelialmarker protein E-cadherin,which may be related to the PI3K/AKT Smad and MMPs signaling pathways.(2)Compared with cisplatin alone group,the combination of resveratrol 50 mg/kg and cisplatin 5 mg/kg increased the inhibition of tumor grow in nude mice with human breast cancer MDA-MB-231 cells xenograft(P < 0.05).The side effects of cisplatin on weight loss and renal function injury in nude mice were significantly improved.The mechanism of action may be related to the PI3K/AKT,Smad,NF-?B,JNK and ERK signaling pathways..In vitro experiments were conducted to observe the effect of inhibiting the proliferation of MDA-MB-231.The combined use of resveratrol 200 ?M and cisplatin IC50 concentration was superior to that of resveratrol alone,and the CI value was 0.92.The combined effect of cisplatin at 16 ?M and 34?M with resveratrol IC50 was better than that of cisplatin alone,with CI values of 0.812 and 0.955.The combination of resveratrol 12.5,25,25 ?M and cisplatin 4 ?M enhanced the inhibitory effect of cell activity,cell migration and invasion.Compared with the resveratrol group,the cell mobility of the groups of 12.5,25,25 ?M resveratrol and 4 M cisplatin decreased by 19.5%,13.4% and 14.9%,respectively.In the MDA-MB-231 cells' EMT model induced by TGF-?1 5ng/m L,the combination of resveratrol and cisplatin can reverse the TGF-?1 induced EMT,and the mechanism may be related to PI3K/AKT,Smad,NF-?b,JNK and ERK.Conclusion: In the nude mouse model of human breast cancer MDA-MB-231 cells xenograft tumor,resveratrol 100 mg/kg can inhibit lung metastasis of breast cancer cells,inhibit the growth of the tumor,has no significant effect on body weight and liver and kidney function of nude mice.Resveratrol 50mg/kg combined with cisplatin5mg/kg played a synergistic and attenuated role in inhibiting tumor growth,increasing the weight of nude mice and improving renal function.In the EMT model of MDA-MB-231 human breast cancer cells in vitro,resveratrol 12.5,25,50 ?M significantly inhibited cell migration and invasion.The combination of resveratrol and cisplatin enhanced the inhibition effect of cell proliferation and produced synergistic effect.The combination of resveratrol 12.5,25,25 ?M and cisplatin 4 ?M enhanced the inhibitory effect on cell activity,cell migration and invasion.The mechanism of action was related to PI3K/AKT,Smad,NF-?b,JNK and ERK signaling pathways.