姜黄素对胃癌MKN-28细胞增殖、迁移的抑制及对miR-26b、Cyclin D1和Cdx2的影响

Experimental study on the effects of curcumin on proliferation,migration and miR-26b,Cyclin D1,Cdx2 in gastric cancer MKN-28 cells

目的探讨姜黄素对胃癌MKN-28细胞增殖、迁移能力的抑制及miR-26b、Cyclin D1和Cdx2的影响。方法选择胃癌细胞MNK-28细胞,分别在0μmol/L、10μmol/L、20μmol/L、30μmol/L浓度姜黄素条件下培养72 h。分别使用CCK-8法和流式细胞术检测细胞活力和细胞凋周期。细胞划痕实验和Trans-well实验被用于检测细胞迁移和侵袭的能力。并分别使用Western blot和RT-qPCR法检测细胞中Cyclin D1和Cdx2蛋白和miR-26b的表达水平。结果不同浓度姜黄素存在的条件下培养48 h和72 h后细胞活力显著低于对照组(P<0.05),而Cur2和Cur3组的细胞活力均显著低于Cur1组(P<0.05)。随着姜黄素浓度的升高,Cur1、Cur2和Cur3组处于G0/G1期的细胞显著升高,并且Cur2和Cur3组处于G0/G1期的细胞比例显著高于Cur1组(P<0.05),而处于G2/M期的细胞比例显著低于Cur1组(P<0.05)。Cur2和Cur3组的细胞迁移率和侵袭率均显著低于对照组和Cur1组(P<0.05)。随着姜黄素浓度的升高,胃癌细胞表达的miR-26b显著升高而Cyclin D1和Cdx2蛋白的水平显著降低(P<0.05)。姜黄素可显著抑制MMP2、MMP9蛋白水平,而促进E-cadherin表达(P<0.05)。结论姜黄素具有抗胃癌细胞增殖、迁移和侵袭的作用,姜黄素可能通过上调miR-26b的表达调节细胞周期相关蛋白和转移相关蛋白的表达发挥抗肿瘤作用。

Objective To investigate the effects of curcumin on proliferation,migration and miR-26b,Cyclin D1,Cdx2 in gastric cancer MKN-28 cells in vitro.Methods The gastric cancer cell MNK-28 cells were cultured for 72 h under the conditions of 0μmol/L,10μmol/L,20μmol/L,30μmol/L of curcumin concentration,respectively.The cell viability and cell cycle were measured using CCK-8 method and flow cytometry,respectively.And the cell scratch assays and Trans-well assays were used to detect the cell migration and invasion.Moreover the expression levels of Cyclin D1,Cdx2 and miR-26b in the cells were detected by Western Blot and RT-qPCR,respectively.Results After 48-hour and 72-hour culture under the conditions of different concentrations of curcumin,the cell viability was significantly lower than that in control group(P<0.05),and the cell viability in Cur2 and Cur3 groups was significantly lower than that in Cur1 group(P<0.05).As the concentration of curcumin increased,the tumor cells numbers at G0/G1 phase in Cur1,Cur2 and Cur3 groups were increased significantly,and the cell proportion at G0/G1 phase in Cur2 and Cur3 groups was significantly higher than that in Cur1 group(P<0.05),however,the cell proportion at G2/M phase was significantly lower than that in Cur1 group(P<0.05).The cell migration rate and invasion rate in Cur2 and Cur3 groups were significantly lower than those in control group and those in Cur1 group(P<0.05).With the increase of curcumin concentration,the expression levels of miR-26b in gastric cancer cells were significantly increased,but the levels of Cyclin D1 and Cdx2 proteins were significantly decreased(P<0.05).The curcumin could significantly inhibit the expression levels of MMP-2 and MMP-9 protein,and promot the expression levels of E-cadherin(P<0.05).Conclusion The curcumin has anti-tumor effects on proliferation,migration and invasion of gastric cancer cells,and curcumin may play the anti-tumor effects by up-regulating the expression levels of miR-26b and regulating the expression levels of cell cycle-associated proteins and metastasis-associated proteins.