Ma Tian-Ze, Piao Zhe, Jin Sheng-Yu, Kwak Yong-Geun
Department of Hematology, Yanbian University Hospital, Yanji, Jilin 133000, P.R. China.
Department of Pharmacology, Chonbuk National University Medical School, Jeonju, Jeonbuk 560-182, Republic of Korea.
Exp Ther Med. 2019 Jan;17(1):649-656. doi: 10.3892/etm.2018.7010. Epub 2018 Nov 23.
The exact cause instigating multiple myeloma (MM) has not been fully elucidated, and the disease has a median survival of 6 months without any treatment. To identify potential biomarkers of MM, serum proteins reflecting alteration in their proteomes were analyzed in 6 patients with MM compared with 6 healthy controls using two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-of flight mass spectrometry. The most notable differentially expressed proteins were validated by immunoblotting and changes in mRNA expression were evaluated by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). A total of 11 differentially expressed protein spots were found. The expression levels of 7 proteins [Immunoglobulin heavy constant µ; proto-oncogene diffuse B-cell lymphoma (DBL2); 26S protease regulatory subunit 4 (P26s4); serum albumin; haptoglobin; and two unknown proteins with isoelectronic point (pI) of 6.41 and molecular weight of 35.4 kDa, and pI of 8.05 and molecular weight of 27.4 kDa, respectively] were downregulated in MM compared with healthy controls. Expression of gel actin-related protein 2/3 complex subunit 1A (ARPC1A); immunoglobulin heavy constant γ 1; fibrinogen α chain (FGA) fragment D; and zinc finger protein 70 were increased in serum of MM patients. Protein expressions of ARPC1A, FGA, P26s4 and DBL2 were measured by immunoblotting in an independent cohort of 12 MM patients and 10 healthy controls. RT-qPCR analysis demonstrated that expression only mimicked protein expression, whereas (encoding P26s4) and (encoding DBL2) did not exhibit significant changes in mRNA expression between control and MM samples. These proteins represent putative serological biomarkers for patients with MM.
引发多发性骨髓瘤(MM)的确切病因尚未完全阐明,且该疾病在未经任何治疗的情况下,中位生存期为6个月。为了鉴定MM的潜在生物标志物,我们采用二维电泳(2-DE)和基质辅助激光解吸/电离飞行时间质谱法,分析了6例MM患者与6例健康对照血清中反映蛋白质组变化的蛋白质。通过免疫印迹法验证了最显著差异表达的蛋白质,并通过逆转录定量聚合酶链反应(RT-qPCR)评估了mRNA表达的变化。共发现11个差异表达的蛋白质斑点。与健康对照相比,MM患者中有7种蛋白质[免疫球蛋白重链恒定区μ;原癌基因弥漫性B细胞淋巴瘤(DBL2);26S蛋白酶调节亚基4(P26s4);血清白蛋白;触珠蛋白;以及两种等电点(pI)分别为6.41、分子量为35.4 kDa和pI为8.05、分子量为27.4 kDa的未知蛋白质]的表达下调。MM患者血清中凝胶肌动蛋白相关蛋白2/3复合体亚基1A(ARPC1A)、免疫球蛋白重链恒定区γ₁、纤维蛋白原α链(FGA)片段D和锌指蛋白70的表达增加。在12例MM患者和10例健康对照的独立队列中,通过免疫印迹法检测了ARPC1A、FGA、P26s4和DBL2的蛋白质表达。RT-qPCR分析表明, 的表达仅与蛋白质表达相似,而 (编码P26s4)和 (编码DBL2)在对照和MM样本之间的mRNA表达未表现出显著变化。这些蛋白质代表了MM患者的潜在血清生物标志物。