Curcumin inhibiting malignant biological behaviors of human neuroblastoma SH-SY5Y cells by up-regulating miR-34a
Objective To explore the effects and possible action mechanism of curcumin on malignant biological behaviors of human neuroblastoma SH-SY5Y cells.Methods The cells viability treated with curcumin(0,6.25,12.5,25,50,100 μmol L-1)was detected by CCK8,and the subsequent treatment concentration of curcumin(0,6.25,12.5,25 μmol·L-1)were screened out.The cells apoptosis,migration,invasion and levels of related proteins were detected by flow cytometry,scratch assay,Transwell assay and Western blot.The expression level of miR-34a in SH-SY5Y cells was detected by qRT-PCR.The SH-SY5Y cells line with low-expression miR-34a(miR-34a inhibitor group),its negative control(NC group)and control group(Control group)were set up.The cell lines in the three groups were treated with curcumin(25 μmol·L-1)and included into curcumin+miR-34a inhibitor group,curcumin+NC group and and curcumin group,respectively.And blank control group was set up.The cells apoptosis,migration and invasion,and levels of related proteins,p-p65 and p-IκBα in each group were detected by the above-mentioned method.Results The survival rate of SH-SY5Y cells treated with curcumin(12.5 μmol·L-1)was significantly lower than that without curcumin treatment(P＜0.05).The apoptosis rate,expressions of Cleaved cas3/cas3,Cleaved cas9/cas9 and E-cadherin,and expression level of miR-34a in curcumin(12.5,25 μmol·L-1)groups were significantly higher than those in curcumin(0 μmol·L-1)group(P＜0.01),while closure rate of scratched cells,number of invasion cells,expressions of N-cadherin and Snail were significantly lower than those in curcumin(0 μmol·L-1)group(P＜0.05).The apoptosis rate,expressions of Cleaved cas3/cas3,Cleaved cas9/cas9 and E-cadherin in curcumin group were significantly higher than those in blank control group(P＜0.01),while closure rate of scratched cells,number of invasion cells,expressions of N-cadherin,Snail,p-p65/p65 and p-IκBα/IκBα were significantly lower than those in blank control group(P＜0.01).The apoptosis rate,expressions of Cleaved cas3/cas3,Cleaved cas9/cas9 and E-cadherin in curcumin+miR-34a inhibitor group were significantly lower than those in curcumin+NC group(P＜0.01),while closure rate of scratched cells,number of invasion cells,expressions of N-cadherin,Snail,p-p65/p65 and p-IκBα/IκBα were significantly higher than those in curcumin+NC group(P＜0.01).Conclusion Curcumin can promote apoptosis of SH-SY5Y cells,inhibit cells migration and invasion,which may be related to up-regulating miR-34a expression and inhibiting NF-κB signaling pathway.
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