Wang Wei, Gu Mei-Feng, Wang Zhi-Fei, Shen Xiang-Min, Zhang Jie, Yang Liang
Department of Neurology, The Second Xiangya Hospital, Central South University, Changsha, 410011, Hunan, China.
Department of Neurosurgery, The Third Xiangya Hospital of Central South University, No. 138, Tongzipo Road, Changsha, 410078, Hunan, China.
Apoptosis. 2021 Oct;26(9-10):548-560. doi: 10.1007/s10495-021-01685-x. Epub 2021 Aug 18.
Schwann cells (SCs) have important roles in supporting and repairing peripheral neurons, and thus have great potential for nerve injury treatment. Adipose tissue-derived stem cells (ADSCs) can be reliably induced to differentiate into SCs. However, the underlying molecular mechanisms are unclear. We explored the roles of MEG3/let-7a-5p/RBPJ axis in the differentiation into SCs from ADSCs. Primary ADSCs were induced to differentiate into SCs by appropriate reagents. ELISA, immunostaining, Western blotting, and qRT-PCR were employed to examine levels of SC-markers such as S100, GFAP, SOX10, p75NTR, GAP43, MPZ, β-NGF, BDNF, and NCAM and let-7 family, MEG3, RBPJ, and Notch signaling related proteins. Dual luciferase assay and RNA immunoprecipitation were performed to validate interactions of let-7a-5p/RBPJ mRNA and MEG3/let-7a-5p. Cultured ADSCs could be induced to differentiate into functional SCs. Let-7a-5p and let-7d-5p were elevated during the differentiation while MEG3 and RBPJ/Notch-signaling were suppressed. Let-7a-5p mimics promoted ADSC differentiation into SCs and up-regulated the levels of SC-related markers including S100, GFAP, SOX10, p75NTR, GAP43, MPZ, β-NGF, and NCAM, while RBPJ or MEG3 overexpression retarded the differentiation and reduced those levels. Let-7a-5p directly targeted RBPJ and MEG3 disinhibited Notch-RBPJ signaling via sponging let-7a-5p. RBPJ overexpression reversed the acceleration of let-7a-5p mimics on SC differentiation while let-7a-5p mimics blocked MEG3-mediated suppression on SC differentiation. Let-7a-5p sponged by MEG3 promotes differentiation of ADSCs into SCs via suppressing Notch signaling by targeting RBPJ. These findings shed light on mechanisms underlying the differentiation of ADSCs to SCs and provide avenues to accelerate the process.
施万细胞(SCs)在支持和修复周围神经元方面发挥着重要作用,因此在神经损伤治疗中具有巨大潜力。脂肪组织来源的干细胞(ADSCs)能够被可靠地诱导分化为SCs。然而,其潜在的分子机制尚不清楚。我们探究了MEG3/let-7a-5p/RBPJ轴在ADSCs向SCs分化过程中的作用。用合适的试剂诱导原代ADSCs分化为SCs。采用酶联免疫吸附测定(ELISA)、免疫染色、蛋白质免疫印迹法(Western blotting)和定量逆转录聚合酶链反应(qRT-PCR)检测SCs标志物如S100、胶质纤维酸性蛋白(GFAP)、SRY(Y染色体性别决定区)-盒转录因子10(SOX10)、低亲和力神经生长因子受体(p75NTR)、生长相关蛋白43(GAP43)、髓鞘蛋白零(MPZ)、β-神经生长因子(β-NGF)、脑源性神经营养因子(BDNF)和神经细胞黏附分子(NCAM)以及let-7家族、MEG3、RBPJ和Notch信号相关蛋白的水平。进行双荧光素酶测定和RNA免疫沉淀以验证let-7a-5p/RBPJ mRNA和MEG3/let-7a-5p之间的相互作用。培养的ADSCs可被诱导分化为功能性SCs。在分化过程中,let-7a-5p和let-7d-5p水平升高,而MEG3和RBPJ/Notch信号被抑制。let-7a-5p模拟物促进ADSCs向SCs分化,并上调包括S100、GFAP、SOX10、p75NTR、GAP43、MPZ、β-NGF和NCAM在内的SCs相关标志物水平,而RBPJ或MEG3过表达则阻碍分化并降低这些水平。let-7a-5p直接靶向RBPJ,MEG3通过吸附let-7a-5p解除对Notch-RBPJ信号的抑制。RBPJ过表达逆转了let-7a-5p模拟物对SCs分化的促进作用,而let-7a-5p模拟物则阻断了MEG3介导的对SCs分化的抑制作用。MEG3吸附的let-7a-5p通过靶向RBPJ抑制Notch信号来促进ADSCs向SCs分化。这些发现揭示了ADSCs向SCs分化的潜在机制,并为加速这一过程提供了途径。