Department of Molecular Cell Biology and Oral Anatomy, Kyushu University Graduate School of Dental Science, Fukuoka 812-8582, Japan.
Department of Periodontology, Faculty of Dental Science, Kyushu University, Fukuoka 812-8582, Japan.
J Cell Sci. 2024 Oct 1;137(19). doi: 10.1242/jcs.261732. Epub 2024 Oct 10.
Non-excitable cells express sodium voltage-gated channel alpha subunit 1 gene and protein (known as SCN1A and NaV1.1, respectively); however, the functions of NaV1.1 are unclear. In this study, we investigated the role of SCN1A and NaV1.1 in human mesenchymal stem cells (MSCs). We found that SCN1A was expressed in MSCs, and abundant expression of NaV1.1 was observed in the endoplasmic reticulum; however, this expression was not found to be related to Na+ currents. SCN1A-silencing reduced MSC proliferation and delayed the cell cycle in the S phase. SCN1A silencing also suppressed the protein levels of CDK2 and AKT (herein referring to total AKT), despite similar mRNA expression, and inhibited AKT phosphorylation in MSCs. A cycloheximide-chase assay showed that SCN1A-silencing induced CDK2 but not AKT protein degradation in MSCs. A proteolysis inhibition assay using epoxomicin, bafilomycin A1 and NH4Cl revealed that both the ubiquitin-proteasome system and the autophagy and endo-lysosome system were irrelevant to CDK2 and AKT protein reduction in SCN1A-silenced MSCs. The AKT inhibitor LY294002 did not affect the degradation and nuclear localization of CDK2 in MSCs. Likewise, the AKT activator SC79 did not attenuate the SCN1A-silencing effects on CDK2 in MSCs. These results suggest that NaV1.1 contributes to the cell cycle of MSCs by regulating the post-translational control of AKT and CDK2.
非兴奋细胞表达钠电压门控通道α亚基 1 基因和蛋白(分别称为 SCN1A 和 NaV1.1);然而,NaV1.1 的功能尚不清楚。在这项研究中,我们研究了 SCN1A 和 NaV1.1 在人骨髓间充质干细胞(MSCs)中的作用。我们发现 SCN1A 在 MSCs 中表达,内质网中大量表达 NaV1.1;然而,这种表达与 Na+电流无关。SCN1A 沉默降低了 MSC 的增殖,并延迟了 S 期的细胞周期。SCN1A 沉默还抑制了 CDK2 和 AKT(此处指总 AKT)的蛋白水平,尽管 mRNA 表达相似,并抑制了 MSCs 中的 AKT 磷酸化。环己酰亚胺追踪实验表明,SCN1A 沉默诱导了 CDK2 但不是 MSCs 中的 AKT 蛋白降解。使用环氧酮、巴弗洛霉素 A1 和氯化铵进行的蛋白水解抑制实验表明,泛素-蛋白酶体系统和自噬和内体-溶酶体系统都与 SCN1A 沉默的 MSCs 中 CDK2 和 AKT 蛋白减少无关。AKT 抑制剂 LY294002 不影响 CDK2 在 MSCs 中的降解和核定位。同样,AKT 激活剂 SC79 也不能减轻 SCN1A 沉默对 MSCs 中 CDK2 的影响。这些结果表明,NaV1.1 通过调节 AKT 和 CDK2 的翻译后控制,促进 MSCs 的细胞周期。