Hinokiflavone Inhibits Growth of Esophageal Squamous Cancer By Inducing Apoptosis via Regulation of the PI3K/AKT/mTOR Signaling Pathway

Abstract

BackgroundGlobally, esophageal cancer ranks as the seventh most common cancer. Esophageal squamous cell carcinoma (ESCC) is one of its major histological types. ESCC accounts for the vast majority of cases in China, and the mortality rate is high. Cisplatin, the standard adjuvant chemotherapy drug for ESCC, has a modest response rate due to the development of drug resistance. Hinokiflavone (HF) is a natural biflavonoid compound with anti-melanoma activity. However, its anti-tumor effect on ESCC and the underlying mechanisms remain largely unknown.MethodsThe ESCC cell lines KYSE150 and TE14 were used. The cell counting kit-8 assay and flow cytometry analysis, along with colony formation, EdU, wound healing, and Transwell migration assays, were performed to assess cell characteristics (viability, migration, invasion, and apoptosis) following treatment with HF. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), western blotting, and molecular docking were used to investigate the pathways potentially modulated by HF. In vivo anti-tumor effects of HF were also investigated using a mouse xenograft model.ResultsOur findings revealed that HF inhibited ESCC cell proliferation. Hoechst 33342 staining, annexin V-FITC/PI staining, and western blotting confirmed that HF causes caspase-dependent apoptosis. KEGG pathway enrichment analysis and western blotting indicated that the PI3K/AKT/mTOR pathway played an important role in the process of HF-induced apoptosis. Furthermore, HF effectively impaired the migration and invasion abilities of KYSE150 cells and downregulated the expression of the matrix metalloproteinases (MMP) MMP2 and MMP9. HF inhibited tumor growth and exhibited minimal toxicity in the organs of the KYSE150 xenograft model.ConclusionThis is the first study to demonstrate the inhibition of ESCC growth and progression by HF. The underlying mechanism is through blocking the PI3K/AKT/mTOR signaling pathway, thereby inhibiting cell proliferation and inducing apoptosis. HF can be used as a complementary/alternative agent for ESCC therapy.

Anti-tumor effect and mechanism of the total biflavonoid extract from S doederleinii on human cervical cancer cells in vitro and in vivo

“…Studies have found that many biflavones play an anti-tumor role by inducing apoptosis and/or autophagic cell death of tumor cells. For example, the biflavone Hinokiflavone suppresses the growth of esophageal squamous cell carcinoma by regulating PI3K/AKT/mTOR signal pathway to induce apoptosis [ 26 ]. Delicaflavone plays an important role in preventing NSCLC, colon cancer and cervical cancer by activating mitochondrial dependent Caspase signal pathway and inhibiting MAPK and PI3K/Akt/mTOR signal pathway [ 27 , 28 ].…”

Proanthocyanidin oligomers extract from hawthorn mediates cell cycle arrest, apoptosis, and lysosome vacuolation on HCT116 cells

“…The excess dye was washed away with PBS after centrifugation. Antifade mounting medium (Beyotime, China) was added for visualization using inverted fluorescence microscope (model number: CKX31, Olympus, Japan) ( Guo et al, 2022 ).…”

Advances in the Anti-Tumor Activity of Biflavonoids in Selaginella

“…As an illustration, almost all C-C biflavonoids have the capacity to regulate MMPs. Hinokiflavone was found to have a similar effect, decreasing the ability of A375 and B16 melanoma cells to invade and migrate, as well as KYSE 150 cells, an esophageal squamous cell carcinoma line, by downregulating MMP-2 and MMP-9 expression [61,88]. Amentoflavone has also been discovered to inhibit invasion and migration of bladder cancer cells by reversing EMT via nuclear factor-kappa B (NF-κB) inactivation and by lowering the expression of MMP-2, MMP-9 and uPA [89].…”

“…Furthermore, phosphatidylinos itol-3-kinase (PI3K) activates protein kinase B (Akt) by phosphorylating Thr308 and Ser473 (p-Akt) under normal physiological circumstances [118,119]. For example, Guo et al [88] found that hinokiflavone may lead to apoptosis in esophageal squamous cancer cells by controlling the PI3K/Akt/mTOR signaling pathway. Wang et al [120] found that amentoflavone induced apoptosis and inhibited glycolysis in glioma cells by targeting miR-124-3p.…”