Silencing CDCA8 Suppresses Hepatocellular Carcinoma Growth and Stemness via Restoration of ATF3 Tumor Suppressor and Inactivation of AKT/β–Catenin Signaling

Abstract

Big data analysis has revealed the upregulation of cell division cycle associated 8 (CDCA8) in human hepatocellular carcinoma (HCC) and its poorer survival outcome. However, the functions of CDCA8 during HCC development remain unknown. Here, we demonstrate in vitro that CDCA8 silencing inhibits HCC cell growth and long-term colony formation and migration through the accumulation of the G2/M phase cell population. Conversely, CDCA8 overexpression increases the ability to undergo long-term colony formation and migration. RNA sequencing and bioinformatic analysis revealed that CDCA8 knockdown led to the same directional regulation in 50 genes (25 down- and 25 upregulated). It was affirmed based on protein levels that CDCA8 silencing downregulates the levels of cyclin B1 and p-cdc2 and explains how it could induce G2/M arrest. The same condition increased the protein levels of tumor-suppressive ATF3 and GADD34 and inactivated AKT/β–catenin signaling, which plays an important role in cell growth and stemness, reflecting a reduction in sphere-forming capacity. Importantly, it was demonstrated that the extent of CDCA8 expression is much greater in CD133+ cancer stem cells than in CD133− cancer cells, and that CDCA8 knockdown decreases levels of CD133, p-Akt and β-catenin and increases levels of ATF3 and GADD34 in the CD133+ cancer stem cell (CSC) population. These molecular changes led to the inhibition of cell growth and sphere formation in the CD133+ cell population. Targeting CDCA8 also effectively suppressed tumor growth in a murine xenograft model, showing consistent molecular alterations in tumors injected with CDCA8siRNA. Taken together, these findings indicate that silencing CDCA8 suppresses HCC growth and stemness via restoring the ATF3 tumor suppressor and inactivating oncogenic AKT/β–catenin signaling, and that targeting CDCA8 may be the next molecular strategy for both primary HCC treatment and the prevention of metastasis or recurrence.

Development and Validation of a Pyroptosis-Related Signature for Predicting Prognosis in Hepatocellular Carcinoma

“…Interestingly, recent studies indicate that NEIL3 contributes to repairing oxidative telomere damage at mitosis, which is crucial for fighting senescence in HCC cells ( Zhao et al, 2021 ). Knockdown of CDCA8 inhibits HCC cell progression by restoring ATF3 tumor suppressor and inactivating AKT/beta-Catenin signaling ( Jeon et al, 2021 ). SLC2A1 was the gene encoding glucose transporter 1 (Glut-1).…”

A Cell Cycle Progression-Derived Gene Signature to Predict Prognosis and Therapeutic Response in Hepatocellular Carcinoma

“…CDCA8 is in relation to unfavorable stage and survival outcomes of HCC [ 33 ]. CDCA8 inhibition weakens HCC growth and stemness through restoring ATF3 and inactivating AKT/ β -catenin axis [ 34 ]. KPNA2 upregulation accelerates HCC progression via enhancement of migration [ 35 ] and proliferation and is indicative of undesirable prognosis [ 36 ].…”

Liquid–Liquid Phase Separation-Related Genes Associated with Tumor Grade and Prognosis in Hepatocellular Carcinoma: A Bioinformatic Study

“… 33 Silencing CDCA8 can suppress HCC growth and stemness. 34 RAD51 appears to influence the onset, progression and drug resistance of various cancers. 35 NCL and CENPA should be explored further to confirm and clarify their involvement in HCC.…”