姜黄素通过调控TUFT1/AKT信号通路抑制肝细胞癌增殖、迁移及侵袭研究

Study on Inhibition of Curcumin on Proliferation,Migration and Invasion of Hepatocellular Carcinoma by Regulating TUFT1/AKT Signaling Pathway

目的:探讨姜黄素对肝细胞癌细胞系HCCLM3体外增殖、迁移及侵袭的影响及潜在分子信号机制。方法:利用不同浓度的姜黄素(0、30、60μmol/L对应为空白对照组、姜黄素处理组1、姜黄素处理组2)处理HCCLM3细胞,利用四甲基偶氮唑蓝(MTT)法检测细胞的增殖情况,利用Transwell法检测细胞的迁移及侵袭能力改变;利用姜黄素(0、60μmol/L)处理HCCLM3细胞,利用qRT-PCR及Western blot法检测细胞内TUFT1表达改变及AKT蛋白磷酸化。结果:MTT实验结果显示,相较于空白对照组,姜黄素处理组1及姜黄素处理组2的HCCLM3细胞增殖能力显著降低(P<0.05)。Transwell迁移实验结果显示,相较于空白对照组,姜黄素处理组1及姜黄素处理组2的HCCLM3细胞迁移能力显著降低(P<0.05)。Transwell侵袭实验结果显示,相较于空白对照组,姜黄素处理组1及姜黄素处理组2的侵袭能力显著降低(P<0.05)。qRT-PCR实验结果显示,相较于空白对照组,姜黄素处理组2的HCCLM3细胞内TUFT1 mRNA表达水平显著降低(P<0.05)。Western blot结果显示,姜黄素处理组2的HCCLM3细胞内TUFT1蛋白表达水平显著降低,AKT蛋白磷酸化显著降低(P<0.05)。结论:姜黄素可能通过抑制TUFT1/AKT信号通路活化,发挥抑制肝细胞癌增殖、迁移及侵袭的作用。

Objective:To discuss the effect of curcumin on proliferation,migration and invasion of HCCLM3 cell line in vitro and its potential molecular mechanism. Methods:In the blank control group,HCCLM3 cells were treated with 0μmol/L of curcumin;in the curcumin group 1,HCCLM3 cells were treated with 30 μmol/L of curcumin;in the curcumin group 2,HCCLM3 cells were treated with 60 μmol/L of curcumin. In the three groups, cell proliferation was detected by MTT method, and cell migration and invasion were detected by Transwell method. In the blank control group and the curcumin group 2, the changes of expression of TUFT1 and phosphorylation of AKT were detected by qRT-PCR and Western blot.Results:The experimental results of MTT showed that,compared with that in the blank control group,the proliferation of HCCLM3 cells in the curcumin group 1 and 2 was significantly decreased(P<0.05). The results of Transwell Migration Assay showed that,compared with that in the blank control group,the migration of HCCLM3 cells in the curcumin group 1 and2 was significantly decreased(P<0.05). The results of Transwell Invasion Assay showed that,compared with that in the blank control group,the invasion of HCCLM3 cells in the curcumin group 1 and 2 was significantly decreased(P<0.05). The results of qRT-PCR showed that,compared with that in the blank control group,the expression level of TUFT1 mRNA in HCCLM3 cells in the curcumin group 2 was significantly decreased(P<0.05). The results of Western blot showed that,in the curcumin group 2,the expression level of TUFT1 protein and the phosphorylation of AKT in HCCLM3 cells were significantly decreased(P<0.05). Conclusion: Curcumin may inhibit the proliferation, migration and invasion of hepatocellular carcinoma by inhibiting the activation of TUFT1/AKT signaling pathway.