The Effect And Possible Mechanism Of Metformin On Mouse Breast Cancer Cell Line4T1

ObjectiveThe objective of this study was to evaluate the effect of metformin and metformincombined with the AMPK inhibitor Compound C on mouse breast cancer cell line4T1about proliferation, observe whether the metformin can kill this cell line, as well as toinvestigate the potential molecular target(s) of metformin-mediated antitumor activity.Methods1. In vitro culture of mouse breast cancer cell line4T1to the logarithm growth periodwere obtained to treated with metformin2.5mmol/L,5mmol/L10mmol/L,20mmol/L,40mmol/L and20μmol/L Compound C respectively and combined with10mmol/Lmetformin for24h,48h and72h;2. CCK-8was used to study the effect of metformin oncell growth with concentration0,2.5,5,10,20,40mmol/L metformin and groups(20μmol/L Compound C+10mmol/L metformin、20μmol/L Compound C)24h,48h and72h;3. Western Blot was used to study the expression of cyclinD1protein after metforminwith concentration0,5,10,20,40mmol/L and groups (20μmol/L CompoundC+10mmol/L metformin、20μmol/L Compound C)24h,48h and72h.Results1. CCK-8detection: Cultured cells were treated with metformin (0mmol/L,2.5mmol/L,5mmol/L,10mmol/L,20mmol/L,40mmol/L) for24h,48h and72h. Mousebreast cancer cell line4T1treated with2.5mmol/L metformin groups24h and48hrespectively, in comparison with control group, there is no significant difference betweenthe two groups, P＞0.05, and treated with72h(2.5mmol/L metformin), compared withcontrol group, P＜0.01, the difference was significant between the two groups; Treatedwith metformin(5mmol/L,10mmol/L,20mmol/L,40mmol/L) for24h,48h and72h, theproliferation of mouse breast cancer cell line4T1was significantly inhibited by metformin in a dose-dependent manner. Meanwhile10mmol/L metformin combined with CompoundC and Compound C treatment alone were no significant difference between the controlgroup.2. Western Blot detection: As the concentration of metformin increased the expressionof cyclinD1protein decreased. Group metformin（10mmol/L） combined with CompoundC was no significant difference between the control group.Conclusions1. Metformin can inhibit the cell proliferation of mouse breast cancer cell line4T1,and induce cell apoptosis.2. Compound C can antagonize the inhibition of metformin on the proliferation ofmouse breast cancer cell line4T1.3. Metformin can activity of the AMPK pathway in mouse breast cancer cell line4T1,and inhibits cyclinD1protein expression in mRNA translation level.