Effects Of Curcumin On Invasion And Expression Of MMP-9,CD44 And E-cadherin Of Mucoepidermoid Carcinoma MEC-1 Cells In Vitro

Objective:To study the effect of curcumin on the invasion ability and the expression of MMP-9,CD44 and E-cadherin in mucoepidermoid carcinoma MEC-1 cells in vitro,so as to provide experimental basis for the clinical treatment of mucoepidermoid carcinoma with curcumin.Methods:Mucoepidermoid carcinoma MEC-1 cell line was used as the research object.After resuscitation,the cells were cultured in vitro,and the logarithmic growth phase cells were taken for experiment.The experimental group was curcumin group,and the control group was established（PBS solution containing 0.1%DMSO）.（1）CCK-8 method was used to measure the proliferation inhibition rate of MEC-1 cells（12h,24h,36h）treated with curcumin（5,10,20,40,80μmo1/L）.The experimental data were sorted and analyzed,and the IC₃₀ concentration of curcumin in 24h was calculated by SPSS software.The curcumin concentrations lower than IC30 of 5,10 and 20μmo1/L were selected as the follow-up experimental concentrations.（2）Transwell assay was used to determine the invasion ability of MEC-1 cells treated with curcumin（5,10,20μmo1/L）for 24h.（3）The mRNA expressions of MMP-9,CD44 and E-cadherin in MEC-1 cells treated with curcumin（5,10,20μmo1/L）for 24 hours were detected by real-time quantitative PCR.（4）Western blot was used to detect the expression of MMP-9,CD44and E-cadherin in MEC-1 cells treated with curcumin（5,10,20μmo1/L）for 24 hours.Results:（1）After 12h,24h and 36h of curcumin treatment,with the increase of curcumin concentration,the proliferation inhibition rate of MEC-1 cells increased gradually（P<0.05）,curcumin inhibited the proliferation of MEC-1 cells in a concentration dependent manner.Under the same concentration of curcumin,the inhibitory effect of curcumin on the proliferation of MEC-1 cells increased with time（P<0.05）,in a time-dependent manner.（2）After curcumin treated MEC-1 cells for 24 hours,the number of MEC-1 penetrating the microporous membrane in curcumin groups was significantly different from that in the control group（P<0.05）.Curcumin could inhibit the invasion ability of MEC-1 cells cultured in vitro.In a certain concentration range,with the increase of curcumin concentration,the inhibitory effect of curcumin on MEC-1 cell invasion was gradually enhanced in a concentration dependent manner.（3）The results of real-time quantitative PCR showed that the relative expressions of MMP-9mRNA and CD44 mRNA in MEC-1 cells were gradually decreased（P<0.05）and the relative expression of E-cadherin mRNA was gradually increased（P<0.05）with the increase of curcumin concentration 24 hours after curcumin treatment.（4）Western blot analysis showed that with the increase of curcumin concentration,the expression of MMP-9 and CD44 protein decreased（P<0.05）,and the expression of E-cadherin increased（P<0.05）.Conclusion:（1）Curcumin could inhibit the proliferation of MEC-1 cells in a time and concentration dependent manner.（2）Within a certain concentration range,curcumin can inhibit the invasion ability of cultured MEC-1 cells in a concentration-dependent manner.（3）The mechanism of curcumin inhibiting the invasion and metastasis of MEC-1 cells may be related to the down-regulation of MMP-9,CD44 and up-regulation of E-cadherin protein expression.