MiR-103/195/15b And LINC00665 Influence The Proliferation And Migration On Cell By Mediating SALL4

Recent studies has shown that mi RNAs and lnc RNAs play an essential role in regulating gene expression.The aberrant expression of mi RNAs and lnc RNAs is associated with the development and progression of cancer.SALL4 is disregulated in glioma, which influences the gliomagenesis. Up to date, the regulation mechanism of mi RNAs on SALL4 is limited in our lab, and the regulation mechanism of lnc RNA on SALL4, however, remains unexplored.In U251 cell line, our former data shows that exogenous expression of mi R-103, mi R-195, mi R-15 b which share the same 5′―seed‖ protion of mi RNA, competitively suppressed cell proliferation, migration and invasion. In this article, we knockdown mi R-103, mi R-195, mi R-15 b in a series of experiments to further investagate their roles.Our results confirmed that down-expression of mi R-103, mi R-195, mi R-15 b could promote the cell proliferation in U87 MG cell through MTT assay and promote the cell migratioin and invasion through Transwell experiments. Besides, down-regulation of mi R-103/195/15 b simultaneously did not generate synergy effect on cell proliferation, migration and invasion, indicating that mi R-103, mi R-195, mi R-15 b impact glioma competitively. In addition, using a dual-luciferase reporter system, SALL4 3′-untranslated region-dependent luciferase activity was increased by mi R-103 inhibitor, mi R-195 inhibitor and mi R-15 b inhibitor. Furthermore, SALL4 level was competitively up-regulated upon mi R-103/195/15 b joint down-expression in glioma cell through immunofluorescence technology.We discovered that SALL4 co-express with LINC00665 by RNAseq database analysis. So this article explored the role of LINC00665 in glioma. First, we detected the level of LINC00665 and found LINC00665 was up-regulated in glioma tissues. Associated with the expression of SALL4 in glioma tissues, we observed a positive correlation between LINC00665 and SALL4 through SPSS software analysis. Second, we declared LINC00665 located in the nucleus by in situ hybrididation. Next, using stable transfection, we generated LINC00665-expressing subclone of U251 cell line. The introduction of LINC00665 into U251, induced cell proliferation, migration and invasion by MTT assay, colony formation assay and Transwell experiments. Cell cycle analysis by PI staining diaplayed, cell cycle was accelerated, and the result obtained by western blot demonstrated the cyclin D1 protein was increased after over-expressing LINC00665. Besides, utilized q RT-PCR, western blot or immunofluorescence analysis, we found that up-regulating of LINC00665 could: 1) suppress mi R-103,mi R-15 b expression significantly, but has no significant difference in mi R-195; 2) promote SALL4 both in m RNA and protein level; 3) increased c-Myc and Tenascin-C m RNA expression levels, and c-Myc protein level was significantly up-regulated.In conclusion, mi R-103/195/15 b targeted to SALL4 and knockdown of mi R-103/195/15 b joint expression could competitively promote cell proliferation, migration and invasion; LINC00665 was up-rgulareded in glioma tissues, it locates in nucleus and overexpression of LINC00665 could inhibit mi R-103, mi R-15 b expression levels, upregulate SALL4 expression and induce cell proliferation, migration and invasion in glioma cell.