CRISPR/Cas9-mediated knockout of NSD1 suppresses the hepatocellular carcinoma development via the NSD1/H3/Wnt10b signaling pathway

Abstract

BackgroundThe NSD family of histone lysine methyltransferases have emerged as important biomarkers that participate in a variety of malignancies. Recent evidence has indicated that somatic dysregulation of the nuclear receptor binding SET domain-containing protein 1 (NSD1) is associated with the tumorigenesis in HCC, suggesting that NSD1 may serve as a prognostic target for this malignant tumor. However, its mechanism in human hepatocellular carcinoma (HCC), the major primary malignant tumor in the human liver, remains unclear. Hence, we investigated how NSD1 regulated HCC progression via regulation of the Wnt/β-catenin signaling pathway.MethodsReverse transcription quantitative polymerase chain reaction (RT-qPCR) and Western blot analysis was performed to identify the expression of NSD1 in HCC cells and clinically obtained tissues. The relationship between NSD1 expression and prognosis was analyzed by Kaplan-Meier survival curve. Further, a NSD1 knockout cell line was constructed by CRISPR/Cas9 genomic editing system, which was investigated in a battery of assays such as HCC cell proliferation, migration and invasion, followed by the investigation into NSD1 regulation on histone H3, Wnt10b and Wnt/β-catenin signaling pathway via ChIP. Finally, a nude mouse xenograft model was conducted in order to assess tumorigenesis affected by NSD1 knockout in vivo.ResultsNSD1 was overexpressed in HCC tissues and cell lines in association with poor prognosis. Knockout of NSD1 inhibited the proliferation, migration and invasion abilities of HCC cells. CRISPR/Cas9-mediated knockout of NSD1 promoted methylation of H3K27me3 and reduced methylation of H3K36me2, which inhibited Wnt10b expression. The results thereby indicated an inactivation of the Wnt/β-catenin signaling pathway suppressed cell proliferation, migration and invasion in HCC. Moreover, these in vitro findings were reproduced in vivo on tumor xenograft in nude mice.ConclusionIn conclusion, the study provides evidence that CRISPR/Cas9-mediated NSD1 knockout suppresses HCC cell proliferation and migration via the NSD1/H3/Wnt10b signaling pathway, suggesting that NSD1, H3 and Wnt10b may serve as potential targets for HCC.

NSD1 stimulated survival and migration of gastric cancer cells through WNT10B

“…In liver cancer, NSD1 was highly expressed, and knockdown of NSD1 could inhibit the cell proliferation, migration and invasion, and inhibited the expression of WNT10B [24]. NSD1 could also affect the progression of HCC through WNT10B [25]. Therefore, we speculated that, in GC, NSD1 may also inhibit the progression of GC through this signaling pathway.…”

“…In fact, the important role of NSD1 in the progression and development of multiple types of tumors has been widely revealed [16]. For example, NSD1 suppressed HCC progression via the NSD1/WNT10B pathway [17]. NSD1 was also highly expressed in pancreatic ductal adenocarcinoma tissues and correlated with its prognosis [8].…”

Advanced therapeutic modalities in hepatocellular carcinoma: Novel insights

“…The HIF‐1α knockout in SMMC 7721 cells significantly inhibited cell invasiveness and migration and prolonged the survival of HCC‐bearing mice. 92 In another report, it was shown that a CRISPR/Cas9‐mediated NSD1 knockout suppressed HCC cell proliferation, migration and invasion via the Wnt/β‐catenin signalling pathway 93 (Figure 2B ).…”

The role of histone methylation in the development of digestive cancers: a potential direction for cancer management

“…177 Similar modifications can be detected in the Wnt10b promoter region in HCC; these modifications activate the Wnt/β-catenin signaling pathway and reinforce cell proliferation, migration, and invasion. 178 Moreover, PRMT6-overexpressing GC cells also acquire invasiveness through direct transcriptional inhibition of PCDH7 by increasing H3R2me2as level. 179 Another study conducted by Li et al demonstrated that H3K27me3 and H3K9me2, mediated directly and indirectly by EZH2, are enriched in the miR-218-2 promoter, thus leading to downregulation of gene expression in PC.…”