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目的探讨褐藻糖胶诱导多发性骨髓瘤细胞PRMI8226凋亡时Wnt/β-catenin通路的变化。方法褐藻糖胶不同浓度(25、50、100、200、400μg·mL-1)分别作用于PRMI8226细胞24、48、72 h,采用MTT法检测PRMI8226细胞增殖情况并求得半数抑制浓度(IC50);以1/2 IC50浓度的褐藻糖胶作用于PRMI8226细胞72 h后,采用流式细胞术检测细胞凋亡率。将细胞分为空白对照组、Wnt通路抑制剂(DKK-1)组(100 ng·mL-1)及褐藻糖胶(25μg·mL-1)组,采用Western blot检测β-catenin、c-myc、bax及caspase 3蛋白表达水平。结果褐藻糖胶对PRMI8226细胞增殖抑制作用呈时间-剂量依赖性增强。PRMI8226细胞经褐藻糖胶25μg·mL-1处理72 h后,其凋亡率(12.22%)明显高于对照组(4.82%)(P<0.05)。Western blot结果显示,与空白对照组比较,两组β-catenin和c-myc表达水平均有显著差异(P<0.05),褐藻糖胶组β-catenin和c-myc表达水平与DKK-1组比较无显著差异(P>0.05);褐藻糖胶组caspase3和bax蛋白表达水平较DKK-1组增加(P<0.05),两组与空白对照组比较表达亦增加,均有显著差异(P<0.05)。结论褐藻糖胶通过抑制Wnt/β-catenin通路的活化,能诱导多发性骨髓瘤PRMI8226细胞凋亡。
Objective To investigate the changes of Wnt / β-catenin pathway in fucoidan induced apoptosis of multiple myeloma cell line PRMI8226. Methods Fucoidan at different concentrations (25,50,100,200,400μg · mL-1) were treated in PRMI8226 cells for 24,48,72 h. The proliferation of PRMI8226 cells was detected by MTT method and the half inhibitory concentration (IC50) The apoptosis rate of PRMI8226 cells treated with fucoidan at a concentration of 1/2 IC50 for 72 h was detected by flow cytometry. The cells were divided into blank control group, Wnt pathway inhibitor (DKK-1) group (100 ng · mL -1) and fucoidan (25 μg · mL -1) group. Western blot was used to detect the expression of β-catenin, , Bax and caspase 3 protein expression levels. Results Fucoidan inhibited the proliferation of PRMI8226 cells in a time-and dose-dependent manner. The apoptosis rate (12.22%) of PRMI8226 cells treated with fucoidan 25 μg · mL-1 for 72 h was significantly higher than that of the control group (4.82%) (P <0.05). The Western blot results showed that the expression levels of β-catenin and c-myc were significantly different between the two groups (P <0.05) compared with the blank control group. The expression of β-catenin and c-myc in the fucoidan group was significantly lower than that in the DKK-1 group (P <0.05). Compared with DKK-1 group, the expression of caspase3 and bax in fucoidan group increased (P <0.05), and the expression of caspase3 and bax in fucoidan group also increased compared with the blank control group (P < 0.05). Conclusion Fucoidan can induce the apoptosis of multiple myeloma PRMI8226 cells by inhibiting the activation of Wnt / β-catenin pathway.