三氧化二砷联合伊曲康唑对多发性骨髓瘤细胞Hedgehog信号通路的影响及抗肿瘤活性实验研究  

Synergistic Inhibitory Effect of Arsenic Trioxide Combined with Itraconazole on Hedgehog Pathway of Multiple Myeloma NCI-H929 Cells

作　　者：黄晓兵[1] 石毅[2] 王春森[1] 王晓冬[1] 陈姣[1] 车菲菲[1] 

机构地区：[1]四川省医学科学院四川省人民医院血液科,四川成都610072 [2]四川省医学科学院四川省人民医院分子生物实验室,四川成都610072

出　　处：《中国实验血液学杂志》2016年第5期1459-1465,共7页Journal of Experimental Hematology

基　　金：四川省卫生厅资助项目(编号090411)

摘　　要：目的:探讨三氧化二砷(ATO)联合伊曲康唑(ITRA)协同抑制多发性骨髓瘤NCI-H929细胞HH信号通路的抗肿瘤作用。方法:应用MTT法检测两药联合对NCI-H929细胞增殖的抑制作用。用多发性骨髓瘤NCI-H929细胞株局部成瘤小鼠模型,观察给药后平均瘤重、瘤重抑制率、肿瘤体积的变化并分析荷瘤小鼠生存情况。ELISA法检测M蛋白,q PCR和Western blot检测Hedgehog信号通路中Ptch、SMO和Gli表达,以及Gli下游靶标基因cyclin D1和BCL-2表达水平变化。结果:ATO联合ITRA与单一给药相比能更显著地抑制NCI-H929细胞增殖。体内实验发现,两药联合应用可更显著地抑制肿瘤生长,降低肿瘤负荷,延迟荷瘤小鼠的生存期。两药联合可显著地下调Hedgehog信号通路下游的关键分子Gli1表达,继而可导致Gli1下游靶标基因cyclin D1和BCL-2等表达水平显著地降低。结论:ATO联合ITRA具有更强地抑制多发性骨髓瘤NCI-H929细胞生长作用。两药协同作用显著地下调Hedgehog信号通路下游的关键分子Gli1表达,而抑制靶标基因的过表达可能是其作用机制之一。Objective：To investigate the antitumor effect of arsenic trioxide（ATO） combined with itraconazole（ITRA） on human multiple myeloma NCI-H929 cells by synergistically inhibiting Hedgehog（HH） signaling pathway.Methods：The inhibitory rate of NCI-H929 cells was assayed by MTT method.Tumor weight,tumor weight inhibition rate,and tumor volume of mouse model with multiple myeloma were examined.The ELISA were appled to detect the M-protein,qPCR and Western blot were used respectively to detect the expression level of Ptch,SMO,Gli and downstream target genes,the survival rate of tumor-bearing mice was analyzed.Results：ATO combined with ITRA significantly inhibited NCI-H929 cell proliferation as compared with a single administration.The combination of ATO and ITRA could synergistically inhibit tumor growth and obviously reduced tumor burden,survival time of tumor-bearing mice was significantly prolonged.qPCR and Western blot results confirmed that the ATO combined with ITRA could significantly down-regulated expression of Gli1,leading to significantly decrease of cyclinD1 and BCL-2 expression levels.Conclusion：ATO combined with ITRA can more strongly suppress the growth of multiple myeloma NCI-H929 cells,as compared with a single administration.The synergistic effect of ATO and ITRA significantly down-regulates expression of Gli1 in HH signaling pathway,moreover the inhibition of target gene overexpression may be one of two drug mechanisms.

关 键 词：伊曲康唑 三氧化二砷 多发性骨髓瘤NCI-H929细胞 HEDGEHOG信号通路 

分 类 号：R733.3[医药卫生—肿瘤] R979.1[医药卫生—临床医学]