Targeting hepatocarcinogenesis model in C56BL6 mice with pan-aurora kinase inhibitor Danusertib

Abstract

Background: To elucidate the expression of Aurora kinases (AURK) and the anticancer effects of pan-aurora kinase inhibitor Danusertib in hepatocarcinogenesis model in C56Bl6 mice.Methods: Thirty mice C56Bl6 were randomly divided into Group A or control, Group B animals who underwent experimental hepatocarcinogenesis with diethylnitrosamine (DEN), and Group C animals with DEN-induced hepatocarcinogenenesis that treated with pan-aurora kinase inhibitor Danusertib. Primary antibodies for immunochistochemistry (IHC) included rabbit antibodies against Ki-67, DKK1, INCENP, cleaved caspase-3, NF-κB p65, c-Jun, β-catenin. Hepatocyte growth factor receptor (C-MET/HGFR) and Bcl-2 antagonist of cell death (BAD) serum levels were determined using a quantitative sandwich enzyme immunoassay technique.Results: Inhibition of AURK reduced the number of DEN-induced liver tumours. Apoptosis and proliferation was very low in both DEN-induced and anti- AURK groups respectively. The hepatocellular adenoma cells of DEN-treated mice uniformly had ample nuclear INCENP whereas in anti- AURK markedly decreased. Expression of β-catenin, NF-kB and c-Jun did not differ in liver tumors of both AURK -depleted and non-depleted mice.Conclusions: Depletion of AURK reduced the number of DEN-induced hepatic tumours. However, their size did not differ significantly between the groups.

<p>HDAC1/2 Inhibitor Romidepsin Suppresses DEN-Induced Hepatocellular Carcinogenesis in Mice</p>

“…HDACi were shown to play a crucial role in cancer progression through regulation of cell proliferation and apoptosis. 33 Although apoptosis has been reported to be a rare event in early HCC lesions in mouse, 29,30 in our study despite the low number of apoptotic cells, morphometric analysis revealed a significant increase of Caspase-3-positive (apoptotic) neoplastic cells in Romidepsin-treated HCC mice, as compared to their non-treated counterparts, which supports a role of Romidepsin in promoting apoptosis. 23 This was further strengthened by the significant elevation of BAD, a major apoptosis regulator, detected in serum of Romidepsin-treated mice, as compared to the HCC untreated controls.…”

“…For testing the effects of Romidepsin on the Wnt/β-Catenin signaling pathway, which is activated in liver cancer, 47,48 we selectively examined catenin (cadherin associated protein), beta 1 (β-catenin) and the dickkopf WNT signaling pathway inhibitor 1 (DKK1). As we have previously described, 30 with β-catenin-specific IHC, DENinduced hepatocellular adenomas are clearly discernible from adjacent non-tumoral liver tissue due to a denser membrane staining. In the present study, this staining pattern was seen not only in adenomas, but in early small-sized HCC lesions as well.…”

“…With Caspase-3-specific IHC we first found that apoptotic cells in HCC lesions are particularly scarce, which matches our previous observations in DEN-induced early hepatocellular carcinogenesis. 29,30 Notably, morphometric counts of Caspase-3-positive cancer cells suggested that HDAC1/2 inhibition upregulates apoptosis in HCC at statistically significant levels (Figure 2). Likewise, using the proliferation marker Ki-67, we found that HCC tumors of Romidepsin-treated mice contained significantly less Ki-67-positive cancer cells compared to their untreated counterparts (Figure 2A).…”

“…In our previous research, we used a DEN mouse model of HCC, which at 10 months after carcinogen challenge, consistently presented with multiple hepatocellular adenomas. 29,30 The same mouse model was used here to evaluate the effect of HDAC blockade on HCC evolution and growth. For that, we treated mice with the HDAC1/2 inhibitor Romidepsin at the critical point of 10 months after DEN challenge and examined their livers 2 months later.…”