Linc-NSC affects cell differentiation, apoptosis and proliferation in mouse neural stem cells and embryonic stem cells in vitro and in vivo

Abstract

Abstract Background Stem cell therapy is a promising therapeutic strategy. Our previous study evaluated tumorigenicity by stereotactic transplantation of mouse neural stem cells (NSCs) and mouse embryonic stem cells (ESCs). When the mice were examined 28 days later, there was no evidence of tumor formation or long-term engraftment in the NSCs transplantation group. In contrast, due to the high proliferative capacity, ESCs transplantation caused tumor formation. Based on transcriptome sequencing, we found that a long intergenic noncoding RNA (named linc-NSC) with unknown structure and function was 1100 times more expressed on NSCs than on ESCs. It is suggested that linc-NSC is negatively correlated with stem cell pluripotency and tumor development, but positively correlated with neurogenesis. Therefore, we further wanted to clarify the role of linc-NSC in NSCs/ESCs in tumor formation and neurogenesis. Methods Whole transcriptome profiling by RNA sequencing and bioinformatics was used to predict the lncRNA widely associated with enhanced tumorigenicity. The expression of linc-NSC was assessed by quantitative real-time PCR. The cell proliferation assay, differentiation assay, immunofluorescence, flow cytometry in vitro, and survival rate and immunofluorescence assays in vivo were performed to investigate the impacts of linc-NSC on tumor formation and neurogenesis in NSCs and ESCs. Results We discovered that after the knockdown of linc-NSC in NSCs, NSCs cultured in vitro and those transplanted into the mouse cortex showed stronger survival ability, enhanced proliferation, and reduced apoptosis, and the opposite result was observed with linc-NSC overexpression in ESCs. Meanwhile, overexpression of linc-NSC in ECSs can induce enhanced apoptosis and differentiation, and inhibit tumorigenesis in vivo, reduction in tumor weight. Conclusions The linc-NSC, a promising gene-edited target, may promote the differentiation of mouse NSCs and inhibit tumorigenesis of mouse ESCs. In other words, knockdown of linc-NSC inhibits NSCs apoptosis in vitro and in vivo, and is unable to trigger tumor formation, revealing a new dimension into the lncRNA of low survival NSCs and providing a prospective gene manipulation target before transplantation. In parallel, overexpression of linc-NSC induces ESCs apoptosis in vitro and in vivo，attenuating the tumorigenicity of ESCs in vivo, although it can’t completely prevent tumor formation.