姜黄素介导微小RNA-199a-3p基因对前列腺癌细胞的影响

目的 探讨姜黄素调控微小RNA-199a-3p(miR-199a-3p)的表达对前列腺癌C4-2细胞增殖、迁移和侵袭的影响.方法 (1)将体外培养的前列腺癌C4-2细胞分为4组:空白组(无药物干预)和低、中、高3个剂量实验组(20,40,80μmol·L-1姜黄素干预),选取对C4-2细胞的增殖抑制最明显姜黄素浓度用于...

Effects of curcumin mediating miR-199a-3p gene on prostate cancer C4-2 cells

Objective To explore the effect of curcumin regulating the expression of miR-199 a-3 p on the proliferation,migration and invasion of prostate cancer C4-2 cells.Methods(1)Prostate cancer C4-2 cells were cultured in vitro and divided into 4 groups:blank group(not intervention)and experimental-L,-M,-H groups,which were intervened with 20,40,80μmol·L^-1 curcumin,respectively.The concentration of curcumin that inhibits the proliferation of C4-2 cells the most obviously was selected to be used in the experiment.(2)Prostate cancer C4-2 cells were cultured in vitro and divided into 3 groups:blank control group,the first transfection group,and the second transfection group.The blank control group only was added liposomes;the first transfection group and the second transfection group were transfected with negative control substance and miR-199 a-3 p inhibitor respectively,and curcumin 40μmol·L^-1 was added.Methyl thiazolyl tetrazoliun was used to detect cell proliferation.Transwell method was used to detect cell migration and invasion.Fluorescence real time quantitative-PCR was used to detect miR-199 a-3 p gene expression(2﹣△△Ctvalue).Western blot method was used to detect matrix metalloproteinase-2(MMP-2),matrix metalloproteinase-9(MMP-9)and Wnt/β-catenin pathwayβ-catenin,Cyclin D1 and c-Myc protein expression(ratios of gray value).Results(1)The cell proliferation rates for 72 h in blank group,experimental-L group,experimental-M group,and experimental-H group were(1.76±0.31)%,(1.52±0.27)%,(0.78±0.16)%and(0.95±0.19)%,with statistically significant difference between experimental-L group,experimental-M,experimental-H group and blank group(all P<0.05).Among them,40μmol·L^-1 curcumin inhibited the proliferation of C4-2 cells most obviously.Therefore,the concentration was selected for subsequent experiments.The number of cell migration in blank group and experimental-M group were(165.48±27.83)and(68.42±16.67),respectively;the number of cell proliferation were(104.62±21.64)and(43.58±10.91),respectively;the expression levels of miR-199 a-3 p gene were 1.03±0.12 and 3.91±0.29,respectively;with statistically significant difference among above indicators between experimental-M group and the blank group(all P<0.05).(2)The cell proliferation rates in blank control group,first transfection group and,second transfection group were(0.95±0.19)%,(0.95±0.27)%and(1.66±0.41)%after 72 h of transfection;the number of cell migration in the three groups were(62.45±12.11),(71.52±15.12)and(138.46±39.49)cells;the number of cell invasion in the three groups were(34.49±8.43),(39.34±9.46)and(81.49±24.16)cells;the expression of MMP-2 protein in the three groups were 0.46±0.11,0.45±0.13 and 0.76±0.17;the expression of MMP-9 protein in the three groups were0.36±0.11,0.38±0.12 and 0.59±0.15;the expression ofβ-catenin protein in the three groups were0.53±0.13,0.49±0.16 and 0.71±0.17;the expression of Cyclin D1 protein in the three groups were0.26±0.09,0.29±0.11 and 0.69±0.15;the expression of c-Myc protein in the three groups were expressions were 0.21±0.06,0.22±0.07 and 0.56±0.12,with statistically significant difference among above indicators between second transfection group and blank control group,or between second transfection group and first transfection group(all P<0.05).Conclusion Curcumin can up-regulate the expression of miR-199 a-3 p gene,thereby inhibiting the proliferation,migration and invasion of C4-2 cells in prostate cancer.