Ryskalin Larisa, Biagioni Francesca, Morucci Gabriele, Busceti Carla L, Frati Alessandro, Puglisi-Allegra Stefano, Ferrucci Michela, Fornai Francesco
Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, Via Roma 55, 56126 Pisa, Italy.
Istituto di Ricovero e Cura a Carattere Scientifico (I.R.C.C.S.) Neuromed, Via Atinense 18, 86077 Pozzilli, Italy.
Cancers (Basel). 2022 Mar 10;14(6):1417. doi: 10.3390/cancers14061417.
Evidence has been recently provided showing that, in baseline conditions, GBM cells feature high levels of α-syn which are way in excess compared with α-syn levels measured within control astrocytes. These findings are consistent along various techniques. In fact, they are replicated by using antibody-based protein detection, such as immuno-fluorescence, immuno-peroxidase, immunoblotting and ultrastructural stoichiometry as well as by measuring α-syn transcript levels at RT-PCR. The present manuscript further questions whether such a high amount of α-syn may be induced within astrocytes, which are co-cultured with GBM cells in a trans-well system. In astrocytes co-cultured with GBM cells, α-syn overexpression is documented. Such an increase is concomitant with increased expression of the stem cell marker nestin, along with GBM-like shifting in cell morphology. This concerns general cell morphology, subcellular compartments and profuse convolutions at the plasma membrane. Transmission electron microscopy (TEM) allows us to assess the authentic amount and sub-cellular compartmentalization of α-syn and nestin within baseline GBM cells and the amount, which is induced within co-cultured astrocytes, as well as the shifting of ultrastructure, which is reminiscent of GBM cells. These phenomena are mitigated by rapamycin administration, which reverts nestin- and α-syn-related overexpression and phenotypic shifting within co-cultured astrocytes towards baseline conditions of naïve astrocytes. The present study indicates that: (i) α-syn increases in astrocyte co-cultured with GBM cells; (ii) α-syn increases in astrocytes along with the stem cell marker nestin; (iii) α-syn increases along with a GBM-like shift of cell morphology; (iv) all these changes are replicated in different GBM cell lines and are reverted by the mTOR inhibitor rapamycin. The present findings indicate that α-syn does occur in high amount within GBM cells and may transmit to neighboring astrocytes as much as a stem cell phenotype. This suggests a mode of tumor progression for GBM cells, which may transform, rather than merely substitute, surrounding tissue; such a phenomenon is sensitive to mTOR inhibition.
最近有证据表明,在基线条件下,胶质母细胞瘤(GBM)细胞的α-突触核蛋白(α-syn)水平很高,与对照星形胶质细胞中测得的α-syn水平相比,过量很多。这些发现通过各种技术得到了一致验证。事实上,通过基于抗体的蛋白质检测方法,如免疫荧光、免疫过氧化物酶、免疫印迹和超微结构化学计量法,以及通过逆转录-聚合酶链反应(RT-PCR)测量α-syn转录水平,都能重复得到这些结果。本论文进一步探讨了在与GBM细胞共培养于跨膜系统中的星形胶质细胞内,是否会诱导产生如此大量的α-syn。在与GBM细胞共培养的星形胶质细胞中,记录到了α-syn的过表达。这种增加伴随着干细胞标志物巢蛋白(nestin)表达的增加,以及细胞形态向GBM样的转变。这涉及一般细胞形态、亚细胞区室以及质膜处的大量卷曲。透射电子显微镜(TEM)使我们能够评估基线GBM细胞内α-syn和巢蛋白的真实含量及亚细胞区室化情况,以及共培养星形胶质细胞内诱导产生的含量,还有超微结构的转变,这种转变让人联想到GBM细胞。雷帕霉素给药可减轻这些现象,它能使共培养星形胶质细胞内与巢蛋白和α-syn相关的过表达及表型转变恢复到幼稚星形胶质细胞的基线状态。本研究表明:(i)与GBM细胞共培养的星形胶质细胞中α-syn增加;(ii)星形胶质细胞中α-syn与干细胞标志物巢蛋白一起增加;(iii)α-syn随着细胞形态向GBM样的转变而增加;(iv)所有这些变化在不同的GBM细胞系中都能重复出现,并且可被雷帕霉素(一种mTOR抑制剂)逆转。本研究结果表明,α-syn确实在GBM细胞中大量存在,并且可能向邻近的星形胶质细胞传递干细胞样表型。这提示了GBM细胞的一种肿瘤进展模式,即GBM细胞可能会转变而非仅仅替代周围组织;这种现象对mTOR抑制敏感。