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Abstract
Background Hepatocellular carcinoma (HCC) is one of the most aggressive malignancies. Previous studies have shown that cancer stem cells play a crucial role in the recurrence and metastasis of tumors. Here, we aim to investigate the role of WD repeat-containing protein 76(WDR76) in promoting HCC progression by enhancing tumor stemness.
Methods We previously established an in vitro hepatocyte differentiation model (IDDF2024-ABS-0312 Figure 1. Establishment of an in vitro hepatocyte differentiation model (A-B)), of which human embryonic stem cells (ES) were induced to be differentiated into definitive endoderm (DE), liver progenitor (LP) cells, and premature hepatocytes (PH). Through deep RNA-sequencing, we found that the WDR76 was highly co-expressed in both embryonic stem cells and liver cancer cells (IDDF2024-ABS-0312 Figure 1. Establishment of an in vitro hepatocyte differentiation model (C)). Bioinformatics analysis of the TCGA dataset and liver cancer clinical specimens has been used to determine the expression of WDR76 in HCC tissues. Functional assays, including spheroid formation, proliferation, apoptosis, migration, and invasion, were performed to evaluate the stemness functions.
Results Analysis of the TCGA database revealed that WDR76 is upregulated in HCC tissues and significantly negatively correlated with prognosis (IDDF2024-ABS-0312 Figure 2. Aberrant expression of WDR76 correlates with poor outcome of HCCS (A-D)). Examination of HCC clinical samples showed that high WDR76 expression correlated with poor differentiation status (IDDF2024-ABS-0312 Figure 2. Aberrant expression of WDR76 correlates with poor outcome of HCCS (E-F)). Also, the expression level of WDR76 was found to be positively correlated with liver cancer stem cell markers (CD133, AFP. IDDF2024-ABS-0312 Figure 2. Aberrant expression of WDR76 correlates with poor outcome of HCCS (I-J)). Functional studies indicated that overexpression of WDR76 could enhance the ability of liver cancer stem cells to form spheroids in 3D culture (IDDF2024-ABS-0312 Figure 3. WDR76 has strong tumorigenic ability (A,B,F)), promote cell growth(Fig.3c)and tolerance to Lenvatinib (IDDF2024-ABS-0312 Figure 3. WDR76 has strong tumorigenic ability (D)), foci formation efficiencies (IDDF2024-ABS-0312 Figure 3. WDR76 has strong tumorigenic ability (E)), migration, invasion (IDDF2024-ABS-0312 Figure 3. WDR76 has strong tumorigenic ability(G-H)), and inhibit apoptosis (IDDF2024-ABS-0312 Figure 3. WDR76 has strong tumorigenic ability(I)). Conversely, knockdown of WDR76 effectively diminished its oncogenic functions (IDDF2024-ABS-0312 Figure 4. Knockdown of WDR76 abolishes its tumorigenicity).
Establishment of an in vitro hepatocyte differentiation model
Aberrant expression of WDR76 correlates with poor outcome of HCCS
WDR76 has strong tumorigenic ability
Knockdown of WDR76 abolishes its tumorigenicity
Conclusions We have demonstrated that WDR76 can promote the progression of HCC by enhancing tumor stemness, and this finding may represent a potential new target for precision therapy of HCC.