Al Othman Aya, Bagrov Dmitry, Rozenberg Julian M, Glazova Olga, Skryabin Gleb, Tchevkina Elena, Mezentsev Alexandre, Durymanov Mikhail
School of Biological and Medical Physics, Moscow Institute of Physics and Technology, Dolgoprudny, Moscow Region, Russia.
Lopukhin Federal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency, Moscow, Russia; Department of Bioengineering, Faculty of Biology, M.V. Lomonosov Moscow State University, Leninskie Gory, Moscow, Russia.
Biochim Biophys Acta Gen Subj. 2024 Jan;1868(1):130522. doi: 10.1016/j.bbagen.2023.130522. Epub 2023 Nov 22.
Activity-regulated cytoskeleton-associated (Arc) protein is predominantly expressed in excitatory glutamatergic neurons of vertebrates, where it plays a pivotal role in regulation of synaptic plasticity. Arc protein forms capsid-like particles, which can encapsulate and transfer mRNA in extracellular vesicles (EVs) between hippocampal neurons. Once glioma cell networks actively interact with neurons via paracrine signaling and formation of neurogliomal glutamatergic synapses, we predicted the involvement of Arc in a process of EV-mediated mRNA transfer between glioma cells.
Arc expression in three human glioma cell lines was evaluated by WB and immunocytochemistry. The properties of Arc protein/mRNA-containing EVs produced by glioma cells were analyzed by RT-PCR, TEM, and WB. Flow cytometry, RT-PCR, and fluorescent microscopy were used to show the involvement of Arc in EV-mediated mRNA transfer between glioma cells.
It was found that human glioma cells can produce EVs containing Arc/Arg3.1 protein and Arc mRNA (or "Arc EVs"). Arc EVs from U87 glioma cells internalize and deliver Arc mRNA to recipient U87 cells, where it is translated into a protein. Arc overexpression significantly increases EV production, alters EV morphology, and enhances intercellular transfer of highly expressed mRNA in glioma cell culture.
These findings indicate involvement of Arc EVs into mRNA transfer between glioma cells that could contribute to tumor progression and affect synaptic plasticity in cancer patients.
活性调节细胞骨架相关蛋白（Arc）主要在脊椎动物的兴奋性谷氨酸能神经元中表达，在突触可塑性调节中起关键作用。Arc蛋白形成衣壳样颗粒，可在海马神经元之间的细胞外囊泡（EVs）中包裹和转运mRNA。一旦胶质瘤细胞网络通过旁分泌信号传导和形成神经胶质瘤谷氨酸能突触与神经元积极相互作用，我们预测Arc参与了胶质瘤细胞之间EV介导的mRNA转运过程。
通过蛋白质免疫印迹（WB）和免疫细胞化学评估三种人胶质瘤细胞系中Arc的表达。通过逆转录聚合酶链反应（RT-PCR）、透射电子显微镜（TEM）和WB分析胶质瘤细胞产生的含Arc蛋白/mRNA的EVs的特性。使用流式细胞术、RT-PCR和荧光显微镜来显示Arc参与胶质瘤细胞之间EV介导的mRNA转运。
发现人胶质瘤细胞可产生含有Arc/Arg3.1蛋白和Arc mRNA的EVs（或“Arc EVs”）。来自U87胶质瘤细胞的Arc EVs内化并将Arc mRNA递送至受体U87细胞，并在其中翻译成蛋白质。Arc过表达显著增加EV的产生，改变EV形态，并增强胶质瘤细胞培养中高表达mRNA的细胞间转运。
这些发现表明Arc EVs参与胶质瘤细胞之间的mRNA转运，这可能有助于肿瘤进展并影响癌症患者的突触可塑性。
