High‑throughput screening identification of a small‑molecule compound that induces ferroptosis and attenuates the invasion and migration of hepatocellular carcinoma cells by targeting the STAT3/GPX4 axis

Abstract

Hepatocellular carcinoma (HCC) is a lethal malignancy. Although considerable efforts have been made in recent years regarding treatments, effective therapeutic drugs for HCC remain insufficient. In the present study, polyphyllin VI was identified as a potential therapeutic drug for HCC by screening natural herbal compounds. The therapeutic effects of polyphyllin VI were assessed using Cell Counting Kit-8, lactate dehydrogenase release and colony formation assays. The occurrence of ferroptosis was determined by assessing lipid peroxidation by reactive oxygen species, malondialdehyde levels, intracellular ferrous iron levels, and the mRNA and protein levels of glutathione peroxidase 4 (GPX4). The migratory and invasive abilities of HCC cells were examined using wound healing and Transwell assays. The results revealed that polyphyllin VI inhibited the proliferation, invasion and metastasis of HCC cells (HCCLM3 and Huh7 cells) by inducing ferroptosis. In addition, through a network pharmacology-based approach and molecular docking analyses, it was found that polyphyllin VI may target the signal transducer and activator of transcription 3 (STAT3). HCC cells were treated with polyphyllin VI or a STAT3 inhibitor (Stattic), both of which exerted similar inhibitory effects on protein expression. Furthermore, immunofluorescence staining revealed that polyphyllin VI significantly inhibited the nuclear translocation of p-STAT3 in HCC cells. Mechanistically, by the overexpression of STAT3, it was confirmed that STAT3 binds to GPX4 and promotes its protein expression and transcription, whereas polyphyllin VI induces ferroptosis by inhibiting the STAT3/GPX4 axis. Subsequently, in vivo experiments revealed that polyphyllin VI inhibited the growth of subcutaneously transplanted tumors. On the whole, findings of the present study suggest that polyphyllin VI inhibits STAT3 phosphorylation, which inhibits GPX4 expression and induces the ferroptosis of HCC cells, eventually inhibiting their invasion and metastasis. These data suggest that polyphyllin VI may be a candidate for the prevention and treatment of HCC.

LASS2 suppresses metastasis in multiple cancers by regulating the ferroptosis signalling pathway through interaction with TFRC

“…Ferroptosis plays a pivotal role in regulating the initiation, invasion, and metastasis of multiple cancers [ 32 , 33 ]. To explore whether LASS2 suppresses tumour cell growth and metastasis through the ferroptosis signalling pathway, the CCK-8 assays, transwell migration and Matrigel invasion assays were performed on multiple tumour cell lines.…”

Polyphyllin VII induces autophagy‐dependent ferroptosis in human gastric cancer through targeting T‐lymphokine‐activated killer cell‐originated protein kinase

“…So far, there is no report of PPVII‐induced ferroptosis. Recently, Huang et al reported that polyphyllin VI of pennogenins induced ferroptosis in the System XC − pathway of hepatocellular carcinoma cells by targeting STAT3 (Huang et al, 2023). In this study, we report that PPVII acts as a direct inhibitor of TOPK to activate autophagy‐dependent iron metabolism‐associated ferroptosis.…”

Targeting epigenetic and posttranslational modifications regulating ferroptosis for the treatment of diseases

“… 535 HCC Polyphyllin VI Phosphorylation STAT3 Polyphyllin VI induces the ferroptosis through inhibiting STAT3 phosphorylation, which inhibits GPX4 expression. 536 HCC Anisomycin Phosphorylation p38MAPK Anisomycin activates p38MAPK to induce ferritinophagy through the phosphorylation of histone H3 on serine 10 (p-H3S10). 537 NSCLC Bufotalin Ubiquitination GPX4 Bufotalin induces ferroptosis by facilitating the ubiquitination and degradation of GPX4.…”

“… 535 Polyphyllin VI induces ferroptosis by inhibiting STAT3 phosphorylation, which inhibits GPX4 expression in HCC. 536 Anisomycin activates p38MAPK to induce ferritinophagy through the phosphorylation of histone H3 on serine 10 (p-H3S10) in HCC. 537 …”