Next‐generation sequencing reveals a novel role of lysine‐specific demethylase 1 in adhesion of rhabdomyosarcoma cells

Abstract

Lysine-specific demethylase 1 (LSD1), a histone lysine demethylase with the main specificity for H3K4me2, has been shown to be overexpressed in rhabdomyosarcoma (RMS) tumor samples. However, its role in RMS biology is not yet well understood. Here, we identified a new role of LSD1 in regulating adhesion of RMS cells. Genetic knockdown of LSD1 profoundly suppressed clonogenic growth in a panel of RMS cell lines, whereas LSD1 proved to be largely dispensable for regulating cell death and short-term survival. Combined RNA and ChIP-sequencing performed to analyze RNA expression and histone methylation at promoter regions revealed a gene set enrichment for adhesion-associated terms upon LSD1 knockdown. Consistently, LSD1 knockdown significantly reduced adhesion to untreated surfaces. Importantly, precoating of the plates with the adhesives collagen I or fibronectin rescued this reduced adhesion of LSD1 knockdown cells back to levels of control cells. Using KEGG pathway analysis, we identified 17 differentially expressed genes (DEGs) in LSD1 knockdown cells related to adhesion processes, which were validated by qRT-PCR. Combining RNA and ChIP-sequencing results revealed that, within this set of genes, SPP1, C3AR1, ITGA10 and SERPINE1 also exhibited increased H3K4me2 levels at their promoter regions in LSD1 knockdown compared to control cells. Indeed, LSD1 ChIP experiments confirmed enrichment of LSD1 at their promoter regions, suggesting a direct transcriptional regulation by LSD1. By identifying a new role of LSD1 in the modulation of cell adhesion and clonogenic growth of RMS cells, these findings highlight the importance of LSD1 in RMS.Additional Supporting Information may be found in the online version of this article.

Single Nuclear RNA Sequencing Highlights Intra-Tumoral Heterogeneity and Tumor Microenvironment Complexity in Testicular Embryonic Rhabdomyosarcoma

“…35 , 36 Collagen, one of the main components of the ECM is an important tissue barrier for cell migration, participates in cell survival and apoptosis, determines cell shape and controls cell differentiation and cell migration. 37 , 38 Collagen VI A1 (COL6A1) is widely present in all connecting tissues. Studies have shown that COL6A1 can trigger signaling pathways related to apoptosis, inflammation, and tumor progression.…”

Lysine specific demethylase 1 is a molecular driver and therapeutic target in sarcoma

“…Recent work has shown that LSD1 instead plays an important role in anchorage independent growth and adhesion in RMS ( 71 ). Knockdown of LSD1 in RMS cells reduces expression of extracellular matrix genes and decreases clonogenic growth, reducing tumor growth, effects that are not assayed in short term growth screens.…”

LINC00675 Suppresses Cell Proliferation and Migration via Downregulating the H3K4me2 Level at the SPRY4 Promoter in Gastric Cancer

“…Evidence has shown that LSD1 mainly targeted histone methylation modification H3K4me2 and predominately induced transcriptional changes. 23 High levels of H3K4me2 at promoter regions may promote gene transcription, whereas low levels of H3K4me2 could inhibit transcription. [40][41][42] Therefore, we performed the ChIP assay for H3K4me2 to further explore the promoter modulation of SPRY4 upon LINC00675 overexpression.…”

“…20,21 LSD1 mediates gene activation and repression via removal of the methyl group (me) from mono-or dimethylated histone H3 at lysine 4 me 1/2 (H3K4me1/2) and lysine 9 (H3K9me1/2), which are endowed with the main specificity for H3K4me2. 22,23 Evidence has shown that LSD1 is upregulated and acts as an oncogene in various kinds of tumors. 24,25 Zhang et al 4 have revealed that LSD1 had upregulated expression in GC and positively contributed to GC development.…”