糖皮质激素和硼替佐米体外干预多发性骨髓瘤细胞BAFF/APRIL mRNA表达的初步探索

Effect of BAFF/APRIL mRNA Expression Induced by Glucocorticoid and Bortezomib in Multiple Myeloma Cells In Vitro

本研究观察糖皮质激素和硼替佐米对U266骨髓瘤细胞株和多发性骨髓瘤(MM)患者骨髓单个核细胞(BMMNC)BAFF/APRIL mRNA表达的影响。分离MM患者BMMNC,对U266骨髓瘤细胞株和BMMNC进行药物干预(单用地塞米松100、200μg/ml,甲基强的松龙100、200μg/ml,硼替佐米0.1μg/ml,以及地塞米松或甲基强的松龙与硼替佐米联用)48小时,收集细胞,进行荧光定量实时PCR检测BAFF/APRIL mRNA表达的水平。采用SPSS 17.0进行统计学分析。结果表明,U266细胞及7例初治的MM患者BMMNC均高表达BAFF/APRIL基因。地塞米松,甲基强的松龙,硼替佐米单独作用于U266细胞或MM患者BMMNC后,BAFF/APRIL基因表达较未干预前降低(p<0.01),其中硼替佐米干预后BAFF/APRIL表达最低(p<0.05)。地塞米松或甲基强的松龙和硼替佐米联用后BAFF/APRIL基因表达较单独干预时低(p<0.01);地塞米松联用硼替佐米时BAFF/APRIL基因表达的抑制强度大于甲基强的松龙和硼替佐米联用的抑制强度(p<0.05)。结论:糖皮质激素和硼替佐米干预后的骨髓瘤细胞BAFF/APRIL基因表达下降,提示糖皮质激素和硼替佐米除了存在已知的糖皮质激素受体和蛋白酶体作用靶点外,可能还存在BAFF/APRIL及其受体这种新的作用靶点。

The study was purposed to detect BAFF/APRIL gene expression changes in bone marrow mononuclear cells（BMMNC） and myeloma cell line U266 after interference with glucocorticoid and bortezomib.After separation of BMMNC from 7 patients with multiple myeloma,BAFF/APRIL mRNA expression in BMMNC and U266 cell line was detected by real-time PCR after treated with dexamethasone 100,200 μg/ml,methylprednisolone 100,200 μg/ml,bortezomib 0.1 μg/ml alone and dexamethasone or methylprednisolone combined with bortezomib respectively for 48 hours.The results showed that U266 cells and BMMNC of untreated MM patients highly expressed BAFF/APRIL genes.When dexamethasone,methylprednisolone or bortezomib was added to U266 cells or BMMNC alone,BAFF/APRIL gene expression decreased as compared with the blank control（p〈0.01）.The inhibiting effect of bortezomib to BAFF/APRIL expression was obviously strong（p〈0.05）.When dexamethasone or methylprednisolone combined with bortezomib,the BAFF/APRIL gene expression further decreased compared with dexamethasone or methylprednisolone alone（p〈0.01）.As compared with the group of methylprednisolone combined with bortezomib,BAFF/APRIL gene expression decreased in dexamethasone combined with bortezomib with a stastically significant difference（p〈0.05）.It is concluded that the expression of BAFF/APRIL gene is down-regulated after bing treated with glucocorticoids and bortezomib,which suggests that besides the glucocorticoid receptor and proteasomes targets,BAFF/APRIL and their rececptor sites may be new targets of glucocorticoids and bortezomib.