Hedgehog信号通路异常对多发性骨髓瘤化疗耐药的影响

发布时间：2018-02-25 22:28

本文关键词： Hedgehog信号通路骨髓瘤耐药　出处：《中国实验血液学杂志》2017年02期 　论文类型：期刊论文

【摘要】：目的:探讨Hedgehog信号过度活化与多发性骨髓瘤耐药的关系。方法:以浓度梯度递增法建立骨髓瘤耐药细胞株RPMI8226/R,实验分为RPMI8226/R,RPMI8226/S,GANT61+RPMI8226/R和GANT61+RPMI8226/S4组,用CCK8法检测4组细胞的增殖抑制率;RT-PCR检测RPMI8226/S和RPMI8226/R耐药前后Gli1,Gli2,Shh,Ihh,Smo和Sufu mRNA表达的变化;Western blot法检测耐药蛋白Cyclin D1,P21和BCL-2及MDR相关信号通路蛋白p-Akt,p-MAPK和STAT3在耐药前后两组细胞的表达。加入不同浓度的GANT61后,Western blot法检测RPMI8226/R,RPMI8226/S组细胞Gli2的表达。结果:Shh、Ihh、Smo、Gli2在RPMI8226/R中表达明显升高,Sufu抑制子则相反,Hedgehog通路下游靶点相关蛋白在RPMI 8226/R中的表达显著高于RPMI8226/S,GANT61体外处理之后,骨髓瘤细胞中Gli2表达量显著减低,GANT61对Gli2表达的减低作用在RPMI8226/R细胞中较RPMI8226/S细胞更为显著,GANT61处理后RPMI8226/R细胞对DOX的敏感性(IC_(50))由7.11±0.061μmol/L升为0.99±0.053μmol/L,相应的耐药指数从5.51降至1.69。结论:Hedgehog信号通路的活化与多发性骨髓瘤的耐药密切相关,GANT61能够阻断Hedgehog信号通路,因此后者可能成为临床克服MM的化疗耐药的一种新的治疗靶点。
[Abstract]:Objective: To investigate the relationship between Hedgehog signal activation and multiple myeloma drug resistance. Method: the concentration gradient method to establish tumor resistant bone marrow cell line RPMI8226/R, the experiment was divided into RPMI8226/R, RPMI8226/S, GANT61+RPMI8226/R and GANT61+RPMI8226/S4 were detected by CCK8 in the 4 groups before and after the inhibition rate of cell proliferation; RT-PCR detection of RPMI8226/S and RPMI8226/R resistant Gli1 Gli2, Shh, Ihh, Sufu, expression of Smo and mRNA; Cyclin D1 Western blot resistance protein detection method, P21 and BCL-2 and MDR signal pathway related protein p-Akt, p-MAPK and STAT3 expression in two groups before and after drug resistance. With different concentrations of GANT61, RPMI8226/R Western blot detection method, the expression of RPMI8226/S cells Gli2. Results: Shh, Ihh, Smo, Gli2 expression in RPMI8226/R was significantly increased, Sufu inhibitor is opposite, the expression of Hedgehog downstream target proteins in RPMI in 8226/R RPMI8226/S was higher than GANT61 in vitro after treatment, significantly reduced the expression of Gli2 in myeloma cells, GANT61 in RPMI8226/R cells than in RPMI8226/S cells decreased expression of Gli2 is more significant, the sensitivity of RPMI8226/R cells to DOX after treatment with GANT61 (IC_ (50)) from 7.11 + 0.061 mol/L + 0.99 + 0.053 mol/L, the resistance index corresponding to 1.69. from the 5.51 conclusion: the Hedgehog signaling pathway is closely related to the activation of drug resistance and multiple myeloma, GANT61 can block the Hedgehog signaling pathway, so the latter may become clinically overcome a new therapeutic target of chemotherapy resistant MM.

【作者单位】：厦门大学附属中山医院血液科;
【基金】：福建省卫计委青年课题(编号2014-2-73) 国家自然科学基金(81172246) 福建省创新医学课题(2014-CXB-42)
【分类号】：R733.3

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