The Influence Of Cinnamaldehyde On The Invasion And Metastasis Of Human Breast Cancer MDA-MB-435S Cells And Its Molecular Mechanism

Objective:This study was designed to explore the influence of cinnamaldehyde on the invasion and metastasis of human breast cancer MDA-MB-435S cells and its molecular mechanism.Methods:①The effect of cinnamaldehyde on the proliferation of human breast cancer MDA-MB-435S cells and the influence of drug concentration and duration on it were tested with MTT method.②The influence of pretreatment of cinnamaldehyde on the adhesion of MDA-MB-435S cells was tested with FN gum and MTT method.③The dose-effect of different concentrations cinnamaldehyde on the invasive ability of MDA-MB-435S cells was observed with transwell chamber model.④The role of miR-27a in the invasive ability of MDA-MB-435S cells,the influence of cinnamaldehyde on miR-27a expression of the cells and its relation with cinnamaldehyde’s inhibiting the invasive ability were investigated with methods of translating miRNA27a mimics/inhibitors with lipofectamin2000,transwell chamber model and RT-PCR.⑤Western blot was performed to test the influence of cinnamaldehyde on protein levels of E-cad, RhoC, stat3and p-stat3in MDA-MB-435S cells.Results:①There were significant inhibiting effects of treatments of cinnamaldehyde with concentrations of5～80μg/ml for24,48and72hours on the proliferation of human breast cancer MDA-MB-435S cells(P<0.01),which were time-dependent and dose-dependent;among them,the highest inhibition rate was81.42%;the half inhibitory concentrations(IC50) of24,48and72hours’groups were29.77,16.60and6.90μg/ml respectively.②There were significant inhibiting effects of cinnamaldehyde with concentrations of20,40and80μg/ml on the adhesion of MDA-MB-435S cells(P<0.05),which were dose-dependent;the highest inhibition rate was79.41%;a rising trend in inhibition rates within30～120min was found in80μg/ml group rather than20and40μg/ml groups.③When treated by cinnamaldehyde with concentrations of20,40and80μg/ml for12h,the count of cells having translocated across the transwell chamber was significantly reduced (P<0.05) and the inhibition rates remained a concentration-dependent manner with cells’ morphology changed in40and80μg/ml groups.④Translated miR-27a mimics and inhibitors,the miR-27a expressions of MDA-MB-435S cells were962.07times and40%as that in control groups respectively;translated miR-27a inhibitors,the count of cells translocating across the transwell chamber successfully was significantly reduced(P<0.05);the miR-27a expression of MDA-MB-435S cells decreased after being treated by cinnamaldehyde with concentrations of20,40and80μg/ml for12h (2-△△Ct=0.56,0.18and0.18respectively);the count of across-the-chamber cells translated miR-27a mimics followed by treatment of cinnamaldehyde for12h was higher than that in groups of treatment of equal concentration cinnamaldehyde following control reagents translating (P<0.05).⑤After treatment of cinnamaldehyde with concentrations of20,40and80μg/ml for24h,the E-cad and RhoC protein grays in MDA-MB-435S cells increased and decreased respectively in a concentration-dependent manner;the stat3protein grays didn’t change in20and40μg/ml groups but decreased in80μg/ml group;the p-stat3protein grays decreased in20,40and80μg/ml groups,among which80μg/ml was most obvious.Conclusion:Cinnamaldehyde suppresses the invasion and metastasis of breast cancer through inhibiting the proliferation, adhesion and invasion of human breast cancer MDA-MB-435S cells.Inhibition of invasive ability of MDA-MB-435S cells by cinnamaldehyde has relation to lowering expression of miR-27a.The effects of different concentrations cinnamaldehyde on MDA-MB-435S cells come true through modulating the expressions of E-cad, RhoC, stat3and p-stat3.