Expression of RAN, ZHX-2, and CHC1L genes in multiple myeloma patients and in myeloma cell lines treated with HDAC and Dnmts inhibitors
Expression of RAN, ZHX-2, and CHC1L genes in multiple myeloma patients and in myeloma cell lines treated with HDAC and Dnmts inhibitors
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DOI：
10.4149/neo_2010_05_482
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发表时间：
2010-01-01
期刊：
影响因子：
3
通讯作者：
Kozubek, S.
中科院分区：
文献类型：
--
作者：
Legartova, S.;Harnicarova-Horakova, A.;Kozubek, S.
真实的时间PCR是研究人类疾病病理生理学中涉及的基因表达的有力工具。最近对RAN（6p 21）、ZHX-2（8q24.3）、CHC 1 L（13q14.3）基因座的研究突出了这些基因在多发性骨髓瘤（MM）预后和治疗应用中的重要性。在这里，我们描述了一个详细的实时PCR方法检测RAN，ZHX-2，和CHC 1 L的表达，这可以应用于临床情况。在健康个体、患有MM的患者和骨髓瘤细胞系MOLP-8的外周血淋巴细胞中研究了这些基因的表达谱。与外周血淋巴细胞和MOLP-8细胞相比，在骨髓瘤患者中观察到RAN、ZHX-2和CHC 1 L的低表达水平。组蛋白去乙酰化酶抑制剂（TSA）能降低CHC 1 L和ZHX-2在MOLP-8细胞中的表达，而RAN在外周血淋巴细胞、对照MOLP-8和TSA或5-氮胞苷处理的MOLP-8细胞中的表达相对稳定。在骨髓瘤患者中，我们观察到选定基因的表达显著降低，但这是患者特异性的。我们的实验说明了在临床实验室进行基因表达研究的新方法和故障排除。
Real time PCR is a powerful tool for studying the expression of genes involved in the pathophysiology of human diseases. Recent studies of the RAN (6p21), ZHX-2 (8q24.3), CHC1L (13q14.3) loci highlight the importance of these genes in multiple myeloma (MM) prognosis and therapeutic applications. Here, we described a detailed Real-Time PCR method for the detection of RAN, ZHX-2, and CHC1L expression, which could be applied in clinical situations. The expression profiles of these genes were studied in peripheral blood lymphocytes of healthy individuals, patients suffering from MM, and in the myeloma cell line, MOLP-8. Low expression levels of RAN, ZHX-2, and CHC1L were observed in myeloma patients, compared with peripheral blood lymphocytes and MOLP-8 cells. An inhibitor of histone deacetylases (TSA) had the ability to decrease expression of CHC1L and ZHX-2 in MOLP-8 cells, while expression of RAN was relatively stable in peripheral blood lymphocytes, control MOLP-8, and TSA- or 5-azacytidine treated MOLP-8 cells. In myeloma patients, we observed significant decreases in the expression of selected genes, but it was patient-specific. Our experiments illustrate new methodological approaches and troubleshooting for conducting gene expression studies in clinical laboratories.