肝癌外泌体<i>LINC ROR</i>对巨噬细胞炎症反应的影响

李雪; 李楠

肝癌外泌体LINC ROR对巨噬细胞炎症反应的影响

李雪,
李楠

LINC ROR from Hepatocarcinoma Cell-derived Exosomes Modulates Inflammation in Human Macrophages

LI Xue,
LI Nan

摘要

摘要: 目的探究肝癌细胞分泌的外泌体中长链非编码RNA（long intergenic non-coding RNA）LINC ROR对巨噬细胞功能的调节作用。方法差速离心分离得到肝癌细胞PLC/PRF/5上清中的外泌体。电镜和纳米颗粒追踪分析仪检测外泌体形态特征和粒径分布。激光共聚焦荧光显微镜观测外泌体与巨噬细胞的空间相对位置。实时荧光定量PCR技术检测LINC ROR 在外泌体中的表达水平。 ELISA法检测巨噬细胞中LINC ROR 小干扰RNA（siRNA）基因沉默后在脂多糖刺激下促炎症因子白细胞介素（IL）-1β的表达水平。结果肝癌细胞分泌的外泌体呈圆状，直径为100 nm左右，同时观测到巨噬细胞能接收肝癌细胞来源的外泌体。相对于正常肝细胞来源的外泌体，肝癌细胞PLC/PRF/5来源的外泌体中高表达LINC ROR。巨噬细胞中LINC ROR基因沉默后，在脂多糖诱导的炎症反应下，巨噬细胞促炎症因子IL-1β的表达水平明显增强。结论肝癌细胞分泌的外泌体中LINC ROR能对邻近巨噬细胞的炎症反应产生调控作用。

Abstract: ObjectiveTo explore the regulatory function of long intergenic non-coding RNA LINC ROR that contained within hepatocarcinoma cells-derived exosomes. MethodsThe physiological appearance of exosomes isolated from PLC/PRF/5 cells was examined by using differential ultracentrifugation and the images were captured by transmission electron microscopy. The internalization of exosomes by THP-1 cells was examined via laser-scanning confocal microscopy. The expression of LINC ROR was analyzed through qRT-PCR. The expression of interleukin （IL）-1β was examined by ELISA. ResultsExosomes were found spherical and the diameters mainly distribute around 100 nm. LINC ROR can be transferred by tumor derived exosomes to macrophages. Exosomes isolated from PLC/PRF/5 cells significantly expressed higher levels of LINC ROR. Suppression of LINC ROR by siRNA significantly increased the release of pro-inflammatory cytokine (IL-1β) in THP-1 cell upon LPS stimulation. ConclusionLINC ROR from hepatocarcinoma cell-derived exosomes plays an important role in regulating the inflammation of macrophages.

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