姜黄素对人胰腺癌PANC-1细胞甲基化转移酶表达影响的体外研究

Experimental study on expression of DNMTs gene in PANC-1 by curcumin

目的探讨姜黄素对人胰腺癌PANC-1细胞甲基化转移酶(DNMTs)基因表达的影响及其作用机制。方法将体外培养的PANC-1细胞分为对照组、吉西他滨组、姜黄素组和联合组。各组分别干预48 h后,采用CCK8法检测人胰腺癌PANC-1细胞的增殖情况,采用RT-PCR法检测DNMTs mRNA表达情况,采用Western blotting法检测DNMT1和Caspase-3蛋白表达水平。结果 1联合组PANC-l细胞增殖抑制率高于姜黄素组和吉西他滨组(P<0.05)。2吉西他滨组、姜黄素组和联合组DNMT1及DNMT3mRNA表达均较对照组降低(P<0.05);联合组低于吉西他滨组和姜黄素组(P<0.05)。3吉西他滨组、姜黄素组和联合组DNMT1蛋白表达均较对照组明显下调(P<0.05,P<0.01),Caspase-3蛋白表达均较对照组明显上调(P<0.01);联合组与吉西他滨组和姜黄素组比较有显著差异(P<0.05)。结论姜黄素可协同吉西他滨促进胰腺癌细胞的凋亡,其机制与调控胰腺癌PANC-1细胞癌基因的去甲基化作用有关。

Objective To investigate the suppression effect on the DNMTs gene by curcumin in pancreatic cancer cell PANC-1 cells and its mechanism.Methods The PANC-1 cell line was divided into control group,gemcitabine group,curcumin group and combination group.After 48 hours intervention CCK8 assay was to detect the proliferation PANC-1 cells.The expression of DNMT1 and DNMT3mRNA were detected by RT-PCR; the expression of DNMT1 and Caspase-3 protein were detected by Western blotting.Results 1 Compared with gemcitabine group and curcumin group,the combination group inhibited the PANC-1 cell significantly higher(P < 0.05).2Compared with the control group,the expression of mRNA of DNMT1 and DNMT3RT-PCR of gemcitabine group,curcumin group and combination group was lower(P < 0.05),while the combination group was lower than that of gemcitabine group and curcumin group(P < 0.05).3Compared with the control group,the expression of DNMT1 of gemcitabine group,curcumin group and combination group was lower(P < 0.05,P < 0.01),while the expression of Caspase-3 was higher(P < 0.01),and there was significant difference between the combination group and the groups of gemcitabine and curcumin(P < 0.05).Conclusion The therapy of combination of gemcitabine and curcumin can synergistically down-regulated the DNMT1 and DNMT3gene expression of mRNA,down-regulated the protein expression of DNMT1,which could via up-regulated the expression of Caspase-3 in the PANC-1 cell line.