BTK与PKCβ在多发性骨髓瘤中表达及调控的信号通路机制研究-东南大学学报医学版

BTK与PKCβ在多发性骨髓瘤中表达及调控的信号通路机制研究

单位：1. 皖南医学院第一附属医院弋矶山医院血液内科, 安徽芜湖 241000;
2. 池州市人民医院血液内科, 安徽池州 247000

分类号：R733.3

出版年·卷·期（页码）：2018·37·第四期（578-583）

摘要：

目的：分析Bruton酪氨酸激酶（BTK）与蛋白激酶Cβ（PKCβ）在多发性骨髓瘤患者肿瘤细胞（MM）和正常人骨髓血淋巴细胞中的表达，探究它们之间的调控关系以及对NF-κB的影响。方法：运用qRT-PCR和Western blotting检测BTK与PKCβ的表达情况。利用siRNA降低BTK的表达，使用抑制剂PCI-32765抑制BTK，在过表达BTK的同时抑制PKCβ，运用qRT-PCR、Western blotting检测PKCβ的变化，Western blotting检测p65核内外分布情况。结果：在30例初诊MM患者标本中，BTK的阳性率为30%，与正常人骨髓血标本相比（阳性率为5%）差异具有统计学意义；PKCβ的阳性率为46.67%，与正常人骨髓血标本相比（阳性率为15%）差异具有统计学意义。在8226细胞中降低BTK的表达或者抑制BTK后，PKCβ的mRNA和蛋白表达下调，而p65的总蛋白表达虽无变化，但是核内的p65下降。过表达BTK可使PKCβ表达上调和p65核内分布增加，但是PKCβ抑制剂LY333531可逆转BTK过表达导致的NF-κB过度激活。结论：BTK与PKCβ在多发性骨髓瘤患者的肿瘤细胞中呈高表达；BTK通过调控PKCβ激活NF-κB。

Objective:To analyze the expression of BTK and PKCβ in tumor cells of patients with multiple myeloma(MM) and normal human bone marrow lymphocytes. To explore the regulatory relationship between BTK and PKCβ and its influence on NF-kappa B. Methods:qRT-PCR and Western blotting was performed to detected the expression of BTK and PKCβ. Using the RNA interference, BTK's expression was reduced; using PCI-32765 BTK was inhibited; while overexpressing BTK in multiple myeloma cancer cells, PKCβ was inhibited. The changes of PKCβ was detected by qRT-PCR and Western blotting, and p65 nuclear distribution was detected by Western blot. Results:In 30 patients with multiple myeloma, BTK's positive rate was 30%, and compared with normal human bone marrow blood samples (positive rate was 5%), the difference was statistically significant. In addition, PKCβ's positive rate was 46.67%, compared with the normal bone marrow blood samples (positive rate was 15%), the difference was statistically significant. The expression of BTK were reduced or inhibited in human multiple myeloma cell line 8226, correspondingly, mRNA and protein expression of PKCβ were decreased. While the total protein expression of p65 did not change, p65 decreased in the nucleus. Overexpression of BTK could increase the expression of PKCβ and increase the distribution of p65 in nucleus, However, PKCβ inhibitor, LY333531, could reverse the over activation of NF-kappa B caused by overexpression of BTK. Conclusion:BTK and PKCβ are highly expressed in tumor cells of patients with multiple myeloma, BTK activates NF-kappa B by regulating PKCβ.

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