Department of Hematology, The Third Affiliated Hospital of Southern Medical University, Guangzhou, China.
Department of Rehabilitation, The First Affiliated Hospital of Jinan University, Guangzhou, China.
Cell Biol Int. 2021 May;45(5):965-975. doi: 10.1002/cbin.11540. Epub 2021 Jan 15.
The bone marrow microenvironment plays an essential role in multiple myeloma (MM) progression. We aimed to explore the alterations of levels of long noncoding RNAs and messenger RNAs (mRNAs), derived from exosomes in peripheral blood, in resistance to bortezomib (Btz) of MM patients. Peripheral blood samples were collected from five Btz-resistant and five Btz-sensitive MM patients. Exosomes in patients' peripheral blood were enriched, and the profiles of long noncoding RNAs (lncRNAs) and mRNAs in exosomes were determined using deep sequencing. Bioinformatics analysis was performed to explore biological function. MTS was employed to determine the viability of Roswell Park Memorial Institute (RPMI) 8226 and LP-1 cells incubated with exosomes derived from Btz-resistant patients. Quantitative polymerase chain reaction (qPCR) was used to evaluate the levels of exosomal FFAR1, SP9, HIST1H2BG, and ITIH2. Incubation with Btz-resistant patient-derived exosomes significantly increased the viability of Btz-treated RPMI 8226 and LP-1 cells in a dose-dependent manner. We identified 482 lncRNAs and 2099 mRNAs deregulated in exosomes of the Btz-resistance group; and 78 mRNAs were enriched in DR-related pathways, including mammalian target of rapamycin, platinum drug resistance, and the cAMP and phosphoinositide 3-kinase-Akt signaling pathways. qPCR results verified the increases in FFAR1 and SP9 and decreases in HIST1H2BG and ITIH2 in Btz-resistant patient-derived exosomes. Moreover, exosomal FFAR1 and SP9 exhibited potential as independent prognostic indicators of survival of MM patients. Our study reveals significant dysregulation of exosomal RNA components in the Btz-resistant group of MM patients as well as several mRNAs that may be used as biomarkers of prognosis of MM patients that are resistant to Btz.
骨髓微环境在多发性骨髓瘤（MM）的进展中起着至关重要的作用。我们旨在探讨多发性骨髓瘤患者对硼替佐米（Btz）耐药时，外周血来源的外体中长链非编码 RNA 和信使 RNA（mRNA）水平的变化。采集了五例 Btz 耐药和五例 Btz 敏感的 MM 患者的外周血样本。从患者的外周血中富集外体，并使用深度测序确定外体中的长链非编码 RNA（lncRNA）和 mRNA 谱。进行生物信息学分析以探索生物学功能。采用 MTS 测定与来自 Btz 耐药患者的外体孵育的 Roswell Park Memorial Institute（RPMI）8226 和 LP-1 细胞的活力。使用定量聚合酶链反应（qPCR）评估外泌体 FFAR1、SP9、HIST1H2BG 和 ITIH2 的水平。与 Btz 耐药患者衍生的外体孵育可显著增加 Btz 处理的 RPMI 8226 和 LP-1 细胞的活力，呈剂量依赖性。我们鉴定了在 Btz 耐药组中外体中失调的 482 个 lncRNA 和 2099 个 mRNA；并且在 DR 相关途径中富集了 78 个 mRNA，包括哺乳动物雷帕霉素靶蛋白、铂类药物耐药和 cAMP 和磷酸肌醇 3-激酶-Akt 信号通路。qPCR 结果验证了 Btz 耐药患者衍生的外体中 FFAR1 和 SP9 的增加以及 HIST1H2BG 和 ITIH2 的减少。此外，外泌体 FFAR1 和 SP9 可作为 MM 患者对 Btz 耐药的生存的独立预后指标。我们的研究揭示了 MM 患者 Btz 耐药组中外体 RNA 成分的显著失调，以及一些可能作为 Btz 耐药的 MM 患者预后生物标志物的 mRNA。
