Objective: To investigate the expression of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) in multiple myeloma (MM) patients, and analyze the effect of doxycycline (DOX) on the expression of MMP-2 and MMP-9 in MM cells.
Methods: The peripheral blood and bone marrow samples of MM patients were collected, and the patients were divided into three groups: newly diagnosed group, remission group and relapsed/refractory group, while the peripheral blood samples of 34 health people and the bone marrow samples of 17 IDA patients were selected as normal control and control group. The levels of MMP-2 and MMP-9 were detected by ELISA. The protein levels of MMP-2 and MMP-9 in H929 cells treated by different concentrations of DOX were analyzed by Western blot. After H929 cells was treated by Akt inhibitor MK-2206 2HCl in combination with DOX, Western blot was used to detect the levels of MMP-2 and MMP-9.
Results: The levels of MMP-2 and MMP-9 in newly diagnosed MM patients were higher than those in control (P<0.05), while for the patients in the remission group were decreased, but still higher than those in control. The levels of MMP-2 and MMP-9 were increased again for the patients in relapsed/refractory group, and showed no significant difference as compared with those in newly diagnosed group. The levels of MMP-2 and MMP-9 could be inhibited by 10 mg/L and 15 mg/L DOX treated by H929 cell. The protein levels of MMP-2 and MMP-9 showed no altered in H929 cells treated by 5 nmol/L MK-2206 2HCl alone. DOX exerted more profound inhibitory effect to MMP-2 and MMP-9 expression in H929 cells when Akt inhibitor MK-2206 2HCl was combined with DOX.
Conclusion: The levels of MMP-2 and MMP-9 are increased in MM patients and related to the disease status of MM. DOX can inhibit the expression of MMP-2 and MMP-9 in MM cells, and antagonizing its activation of Akt signaling pathway can further enhance the inhibitory effect.
题目: 多西环素对多发性骨髓瘤中MMP-2、MMP-9表达的影响.
目的: 研究多发性骨髓瘤（MM）患者基质金属蛋白酶-2（MMP-2）和基质金属蛋白酶-9（MMP-9）的表达及临床意义，并进一步研究多西环素（DOX）对MM细胞MMP-2、MMP-9表达的影响.
方法: 收集MM患者外周血浆及骨髓标本，将患者分为新诊断组、缓解组和难治复发组，另选取34例我院健康体检者的外周血浆为正常对照组，17例我院血液科就诊的缺铁性贫血患者的骨髓为对照组，ELISA法检测MMP-2、MMP-9的表达；不同浓度DOX处理骨髓瘤细胞株H929一定时间后，Western blot法检测MMP-2、MMP-9的表达；联合使用Akt抑制剂MK-2206 2HCl和DOX处理H929一定时间后，Western blot法检测MMP-2、MMP-9表达的变化.
结果: 新诊断组患者外周血浆和骨髓中MMP-2、MMP-9的表达水平较正常对照组均明显增高（P<0.05），治疗后缓解组患者MMP-2、MMP-9的表达水平较新诊断组下降，但仍高于正常对照组，而难治复发组患者MMP-2、MMP-9的表达水平再次升高，与新诊断组无明显差异；DOX 10和15 mg/L处理H929细胞能明显抑制MMP-2、MMP-9的表达；5 nmol/L的MK-2206 2HCl单独处理H929细胞对MMP-2、MMP-9的表达无明显影响，但是与10 mg/L的DOX联合处理H929细胞可增强DOX对MMP-2、MMP-9 蛋白的抑制作用.
结论: MM患者MMP-2、MMP-9的表达增高，其表达水平与骨髓瘤疾病状态相关。DOX可抑制MM细胞MMP-2、MMP-9的表达，拮抗DOX对Akt信号通路的激活可进一步增强这一抑制作用.
Keywords: MMP-2; MMP-9; doxycycline; multiple myeloma; serine/threonine kinase Akt.