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[目的]通过体外实验观察姜黄素对5-Fu耐药细胞株(5FUR)中NKD2、TET1及Caspase-3表达水平的影响。[方法 ]运用Western blot、Real-time q PCR检测不同浓度姜黄素对5FUR细胞及转染TET1 sh RNA后5FUR细胞内NKD2、Caspase-3蛋白和m RNA表达的影响。通过重亚硫酸测序(BSP)的方法,观察姜黄素对NKD2基因启动子去甲基化的干预情况。[结果]经过5-Fu培养而得到5FUR的IC50为(86.91±3.61)μg/ml(P<0.001)。HCT-116原代细胞5-Fu的IC50为(12.64±1.52)μg/ml,姜黄素对其增殖的抑制率为38.34%(P<0.001),姜黄素能显著上调5FUR细胞内NKD2、TET1的表达,且呈剂量依赖性。TET1转染敲低后检测发现5FUR细胞中的NKD2及Caspase-3的表达受到明显的抑制。[结论]姜黄素能从体外明显抑制结肠癌细胞5FUR的生长,其作用机制可能通过去甲基化调控蛋白TET1促使抑制基因NKD2基因启动子发生去甲基化,从而上调NKD2的表达,最终起到抑制肿瘤生长的作用。
[Objective] To observe the effect of curcumin on the expression of NKD2, TET1 and Caspase-3 in 5-Fu drug resistant cell line (5FUR) in vitro. [Methods] The effects of different concentrations of curcumin on the expression of NKD2, Caspase-3 protein and m RNA in 5FUR cells and 5FUR transfected with TET1 sh RNA were detected by Western blot and Real-time q-PCR. The effect of curcumin on the demethylation of NKD2 promoter was observed by bisulfite sequencing (BSP). [Result] The IC50 of 5FUR obtained by 5-Fu culture was (86.91 ± 3.61) μg / ml (P <0.001). The IC50 of HCT-116 primary cell 5-Fu was (12.64 ± 1.52) μg / ml, curcumin inhibited the proliferation of the cells by 38.34% (P <0.001). Curcumin significantly increased the NKD2, TET1 Expressed in a dose-dependent manner. TET1 transfection knockdown detected 5FUR cells in the NKD2 and Caspase-3 expression was significantly inhibited. [Conclusion] Curcumin can significantly inhibit the growth of colon cancer cell 5FUR in vitro. The mechanism may be that the demethylation of NKD2 gene promoter may be induced by the demethylating regulatory protein TET1, and thus the expression of NKD2 may be increased To inhibit the role of tumor growth.