Emodin inhibits invasion and migration of hepatocellular carcinoma cells via regulating autophagy-mediated degradation of snail and β-catenin

Abstract

Background Previous studies reported that emodin extracted from Rheum palmatum L. exerts antiproliferation and antimetastatic effects in a variety of human cancer types. However, the role of emodin in hepatocellular carcinoma (HCC) remain unknown. Methods EdU and colony formation assays were performed to evaluate the effects of emodin on proliferation. The mobility capacities of HCC treated with emodin were evaluated using wound healing assay. Transwell invasion and migration assays were performed to evaluate anti-migratory and anti-invasive effects of emodin on HCC. Annexin V-FITC/PI was performed to analyze the apoptosis. PI stain was performed to analyze cell cycle. RNA sequencing technology was used to identify the differentially expressed genes (DEGs) induced by emodin in HCC. The impact of emodin on autophagic flux in HepG2 cells was examined by mCherry-GFP-LC3 analysis. Western blot was used to assess the protein expressions of epithelial-mesenchymal transition (EMT), autophagy, PI3K/AKT/mTOR and Wnt/β-catenin signaling pathway. Results We found that emodin inhibited the growth of HepG2 cells in a dose- and time-dependent manner. In addition, emodin inhibited cell proliferation, induced S and G2/M phases arrest, and promoted apoptosis in HepG2 cells. The migration and invasion of HepG2 cells were also suppressed by emodin. Enrichment analysis revealed that DEGs involved in cell adhesion, cancer metastasis and cell cycle arrest. Moreover, western bolt results show that emodin-induced autophagy promotes Snail and β-catenin degradation. We also found that blocking autophagic flux after emodin treatment caused EMT reversal. Furthermore, the PI3K agonist Y-P 740 significantly reversed the phosphorylation levels of GSK3β and mTOR. These results indicated that emodin induced autophagy and inhibited the EMT in part through suppression of the PI3K/AKT/mTOR and Wnt/β-catenin pathways. Conclusion Our study indicated that emodin inhibited cell metastasis in HCC via the crosstalk between autophagy and EMT.

Combination of ethyl acetate fraction from Calotropis gigantea stem bark and sorafenib induces apoptosis in HepG2 cells

“…migrated rate (%) = (width of initial wound—width of remaining wound) / width of initial wound × 100 [ 32 , 33 ].…”

Natural products targeting macroautophagy signaling in hepatocellular carcinoma therapy: Recent evidence and perspectives

“…Its mechanism involves the inhibition of the PI3K/AKT signaling pathway, facilitating the degradation of β‐catenin protein along with accumulated LC3 protein, thereby blocking the Wnt signaling pathway. This dual action inhibits the process of epithelial–mesenchymal transition (EMT) in HepG2 cells (Qin et al, 2022). In a study conducted by Zhang et al, Tetrandrine was observed to exert an inhibitory effect on the migration of HCC cells, which is closely associated with the induction of autophagy in human HCC cells.…”

Anti-Cancer Potential of Phytochemicals: The Regulation of the Epithelial-Mesenchymal Transition

“…In hepatocellular carcinoma cells, emodin treatment activated autophagy and promoted autophagic flux, simultaneously reversed EMT, and indicated the correlation between EMT and autophagy. Both EMT and autophagy regulation were revealed to be attributed to the PI3K/AKT/mTOR pathway [ 114 ]. By regulating EMT, emodin could reduce cancer cell resistance to chemotherapeutic drugs, such as gemcitabine resistance in pancreatic cancer cells and doxorubicin resistance in small-cell lung cancer cells [ 115 , 116 ].…”