葡萄籽原花青素下调miR-27a表达抑制胰腺癌细胞生长

Grape seed proanthocyanidins extract inhibits pancreatic cancer cell growth through down-regulation of miR-27a expression

目的:探讨葡萄籽原花青素(grape seed proanthocyanidins extract,GSPE)抑制胰腺癌细胞生长的作用及其可能机制。方法:体外培养人胰腺癌As PC-1细胞,分别采用MTT,Annexin V-FITC/PI双染,Transwell迁移实验等方法检测GSPE对As PC-1细胞的增殖抑制、诱导凋亡及抑制细胞迁移的作用;mi RNA real-time RT-PCR和Western印迹检测GSPE对As PC-1细胞mi R-27a及其下游基因FOXO1表达的影响。转染mi R-27a抑制剂,检测GSPE通过下调mi R-27a调控As PC-1细胞生长、迁移的作用。结果:不同浓度(25,50,75,100μg/m L)GSPE对As PC-1细胞的增殖抑制率分别为8.36%,17.36%,50.42%和60.73%,GSPE≥50μg/m L时,增殖抑制率均显著高于对照组(P<0.05)。对照组及25,50,75μg/m L GSPE组的细胞早期凋亡率依次分别为2.5%,20.0%,42.3%,64.3%。75μg/m L GSPE组的细胞迁移率低于对照组,细胞内mi R-27a的表达亦显著低于对照组(P<0.01)。与对照组相比,mi R-27a抑制剂组、GSPE组、mi R-27a抑制剂+GSPE组的细胞增殖抑制率均显著升高(P<0.01),细胞迁移能力均显著降低(P<0.01),FOXO1蛋白表达增加;且共同处理组上述作用最明显,高于两者单独处理组。结论:GSPE可能通过下调mi R-27a表达来抑制胰腺癌As PC-1细胞生长和迁移。

Objective: To explore the effect of grape seed proanthocyanidins extract(GSPE) on the growth of pancreatic cancer cells and the underlying mechanisms. Methods: The pancreatic cancer As PC-1 cells were cultured in vitro. The effects of GSPE oncell proliferation, apoptosis and migration were analyzed by MTT, Annexin V-FITC/PI and Transwell migration assay, respectively. The expression of mi R-27 a and FOXO1 in As PC-1 cells was determined by real-time RT-PCR and Western blot, respectively. The miR-27 a inhibitors were applied to verify the role of mi R-27 a in mediation of GSPE effects.Results: GSPE inhibited cell growth in a dose-dependent manner. This inhibitory effect was significant when the dosage of GSPE was more than 50 μg/m L(P<0.05 vs control). GSPE also could induce apoptosis and inhibit cell migration. MiR-27 a expression was notably down-regulated when the dosage of GSPE was 75 μg/m L(P<0.01 vs control). Compared with the control group, cell proliferation inhibition was significantly increased in the mi R-27 a inhibitor group, the GSPE group and the miR-27 a inhibitor plus GSPE group(P<0.01), while cell migration was significantly decreased(P<0.01). Compared with the GSPE or the miR-27 a inhibitor group, the growth and migration inhibitory effects in the miR-27 a inhibitor plus GSPE group were more obviously(P<0.01). Both GSPE and miR-27 a inhibitor alone could up-regulate FOXO1 expression. But these effects were more apparent when they are applied in combination.Conclusion: GSPE inhibites AsP C-1 cells' growth and migration partly through down-regulation of mi R-27 a expression.