Hydroxychloroquine (HCQ) Modulates Autophagy and Oxidative DNA Damage Stress in Hepatocellular Carcinoma to Overcome Sorafenib Resistance via TLR9/SOD1/hsa-miR-30a-5p/Beclin-1 Axis

Abstract

Sorafenib is used for treating advanced hepatocellular carcinoma (HCC), but some patients acquire sorafenib resistance. We investigated the mechanisms underlying acquired sorafenib resistance in HCC cells and targeted them to re-sensitize them to sorafenib. In silico analysis indicated that toll-like receptor (TLR)-9 was significantly overexpressed, and that miRNA (hsa-miR-30a-5p) was downregulated in sorafenib-resistant HCC cells, which modulated HCC cell proliferation, oxidative stress, and apoptosis. TLR9 overexpression increased HCC cell proliferation, whereas TLR9 inhibition from hydroxychloroquine (HCQ) decreased HCC cell proliferation, tumor growth, oxidative stress marker (SOD1), and the formation of autophagosome bodies (reduced ATG5 and Beclin-1 expression). Moreover, HCQ treatment reduced epithelial–mesenchymal transition, leading to decreased clonogenicity, migratory ability, and invasiveness. HCQ targeted and reduced the self-renewal capacity phenotype by inhibiting tumorsphere generation. Both in vitro and in vivo results demonstrated the synergistic effect of the HCQ–sorafenib combination on sorafenib-resistant HCC (Huh7-SR) cells, increasing their sensitivity to treatment by modulating TLR9, autophagy (ATG5 and Beclin-1), oxidative stress (SOD1), and apoptosis (c-caspase3) expression and thus overcoming the drug resistance. This study’s findings indicate that TLR9 overexpression occurs in sorafenib-resistant HCC cells and that its downregulation aids HCC suppression. Moreover, HCQ treatment significantly increases sorafenib’s effect on sorafenib-resistant HCC cells.

Autophagy-Related Chemoprotection against Sorafenib in Human Hepatocarcinoma: Role of FOXO3 Upregulation and Modulation by Regorafenib

“…Therefore, we can conclude that instead of an anti-tumor role, autophagy is playing a cytoprotective action in our sorafenib-resistant HCC in vitro models that permits sorafenib-mediated cell death evasion. These findings are also supported by a study where administration of hydroxychloroquine modulates autophagy and oxidative stress to surpass sorafenib resistance in Huh7 cells displaying a resistant phenotype [50]. Similarly, chloroquine promoted the anti-tumor effect of sorafenib in vitro and in vivo [51]; while a combination of sorafenib plus 3-methyladenine, an inhibitor of autophagy at early phases, also enhanced the apoptotic rate in HCC [52].…”

Hydroxychloroquine treatment on SARS-CoV-2 receptor ACE2, TMPRSS2 and NRP1 expression in human primary pterygium and conjunctival cells

“…Hydroxychloroquine has been reported to induce oxidative DNA damage in mouse embryonic fibroblasts and human hepatocellular carcinoma cells ( Besaratinia et al, 2021 ; Chen et al, 2021 ). We aimed to evaluate the oxidative stress status of pterygium and conjunctival cells after hydroxychloroquine treatment by determining the expression of SOD2 (an oxidative stress marker ( Yang et al, 2019b ); and the cellular ROS levels.…”

“…Hydroxychloroquine would induce cellular oxidative stress ( Besaratinia et al, 2021 ; Chen et al, 2021 ). We confirmed that hydroxychloroquine could increase cellular ROS level ( Fig.…”

Expression Profiles and Prognostic Value of FABPs in Colorectal Adenocarcinomas

“…Activation of the Wnt/β-catenin pathway plays an important role in human CRC tumorigenesis [83]. Moreover, our very recent study identified that FABP6 is the Wnt signaling pathway molecule which may indirectly interact with β-catenin [84]. Therefore, the correlations among FABP6, miR-320, and the Wnt/β-catenin pathway in CRC need to be further explored.…”