下调TPD52基因表达通过Wnt信号通路对子宫肌瘤细胞活力、凋亡及TNF-α和IL-6表达的影响

目的:探讨下调肿瘤蛋白D52(TPD52)基因表达通过Wnt信号通路对子宫肌瘤细胞活力、凋亡及肿瘤坏死因子α(TNF-α)和白细胞介素6(IL-6)表达的影响。方法:将针对TPD52的特异性si RNA(si-TPD52)及其阴性对照转染原代培养的子宫肌瘤细胞,并加入Dickkopf-1(DKK1)作为Wnt信号通路抑制剂,转染48 h后,用Western blot检测TPD52及Wnt信号通路关键分子β-catenin和相关靶蛋白cyclin D1和survivin的蛋白表达; MTT及流式细胞术分别检测细胞活力和凋亡率的变化; RT-qPCR检测TNF-α和IL-6的m RNA表达。结果:TPD52在转染si-TPD52的子宫肌瘤细胞的表达显著低于空白对照组和阴性对照组(P 0. 05);与阴性对照组比较,si-TPD52组细胞活力明显受到抑制,凋亡率增加,TNF-α的m RNA表达降低,IL-6的m RNA表达升高,β-catenin、cyclin D1和survivin的蛋白表达降低(P 0. 05);加入DKK1后si-TPD52对细胞活力和凋亡的影响更明显。结论:下调TPD52基因表达可通过Wnt信号通路抑制子宫肌瘤细胞生长和诱导凋亡,同时下调TNF-α和上调IL-6表达。

Effect of down-regulation of TPD52 expression on viability and apoptosis of uterine fibroid cells and expression of TNF-α and IL-6 through Wnt signaling pathway

AIM: To investigate the effect of down-regulation of tumor protein D52 (TPD52) gene expression on the viability and apoptosis of uterine myoma cells and the expression of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) through Wnt signaling pathway.METHODS: TPD52 specific siRNA and its negative control were transfec-ted into primary uterine fibroid cells for 48 h. The protein expression of TPD52 and Wnt signaling pathway-related molecules, β-catenin, cyclin D1 and survivin, was determined by Western blot. The cell viability and apoptotic rate were analyzed by MTT assay and flow cytometry. The mRNA expression of TNF-α and IL-6 was detected by RT-qPCR.RESULTS: The expression of TPD52 in the uterine fibroid cells transfected with si-TPD52 was significantly lower than that in blank control group and negative control group (P<0.05). Compared with negative control group, the cell viability was significantly inhibited in si-TPD52 group, the apoptotic rate was increased, the mRNA expression of TNF-α was decreased, IL-6 mRNA level was increased, and the protein levels of β-catenin, cyclin D1 and survivin were decreased (P<0.05). More obvious effects of si-TPD52 transfection on the viability and apoptosis of uterine myoma cells were observed after combined with Dickkopf-1 (DKK1), an inhibitor of Wnt signaling pathway.CONCLUSION: Down-regulation of TPD52 gene expression inhibits the viability and induces apoptosis of uterine fibroid cells. It also down-regulates the expression of TNF-α and up-regulates IL-6 expression. These effects were related to inhibition of Wnt signaling pathway.