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目的观察鱼藤素对人多发性骨髓瘤细胞株RPMI-8226细胞增殖和凋亡的影响及其对核受体共激活因子-3(SRC-3)的调控作用,研究鱼藤素诱导凋亡作用与其调节SRC-3的相互关系。方法采用MTT法检测细胞增殖活性,Annexin-VFITC/PI双标法及Hoechst33258染色法分析细胞凋亡的改变,RT-PCR法检测鱼藤素对RPMI-8226细胞内SRC-3基因表达的影响;免疫组化法检测鱼藤素对RPMI-8226细胞SRC-3蛋白的影响和分布情况。结果鱼藤素能明显抑制RPMI-8226细胞的增殖,其抑制作用呈时间、剂量依赖性,其作用24h的IC50为(54.55±0.40)nmol/L。此外,鱼藤素具有较强的诱导RPMI-8226细胞凋亡的效应,25nmol/L即能诱导(17.04±0.73)%的RPMI-8226细胞发生凋亡;当药物浓度达到100nmol/L时,总凋亡率达(63.57±1.36)%。在RPMI-8226细胞中SRC-3高表达,且主要分布于细胞核,经鱼藤素干预后,SRC-3的mRNA和蛋白表达水平明显下降。结论鱼藤素能明显抑制RPMI-8226细胞的增殖,并诱导其凋亡。而鱼藤素诱导的SRC-3表达量的下调可能参与了其诱导凋亡作用。
Objective To observe the effects of deguelin on proliferation and apoptosis of human multiple myeloma RPMI-8226 cells and its regulation on nuclear receptor co-activator-3 (SRC-3), and to investigate the effects of deguelin on apoptosis Role and its regulation of SRC-3 correlation. Methods MTT assay was used to detect cell proliferation. Annexin-VFITC / PI double staining and Hoechst33258 staining were used to analyze the changes of cell apoptosis. RT-PCR was used to detect the effect of deguelin on SRC-3 gene expression in RPMI-8226 cells. Immunohistochemistry was used to detect the effect and the distribution of deguelin on SRC-3 protein in RPMI-8226 cells. Results deguelin significantly inhibited the proliferation of RPMI-8226 cells in a time-and dose-dependent manner. IC50 was (54.55 ± 0.40) nmol / L for 24 h. In addition, deguelin had a stronger effect on inducing RPMI-8226 cell apoptosis, and 25nmol / L induced apoptosis of RPMI-8226 cells (17.04 ± 0.73)%. When the drug concentration reached 100nmol / L, The apoptosis rate reached (63.57 ± 1.36)%. SRC-3 was highly expressed in RPMI-8226 cells and mainly distributed in the nucleus. The mRNA and protein expression of SRC-3 were significantly decreased after intervention of deguelin. Conclusion deguelin can significantly inhibit the proliferation of RPMI-8226 cells and induce its apoptosis. The down regulation of deguelin-induced SRC-3 expression may be involved in the induction of apoptosis.