The antitumor effects of herbal medicine Triphala on oral cancer by inactivating PI3K/Akt signaling pathway: based on the network pharmacology, molecular docking, in vitro and in vivo experimental validation.

BACKGROUND: This research aimed to investigate the anti-tumor effects and underlying genetic mechanisms of herbal medicine Triphala (TRP) in oral squamous cell carcinoma (OSCC). METHODS: The target genes of Triphala (TRP) in oral squamous cell carcinoma (OSCC) were identified, and subsequent functional enrichment analysis was conducted to determine the enriched signaling pathways. Based on these genes, a protein-protein interaction network was constructed to identify the top 10 genes with the highest degree. Genes deregulated in OSCC tumor samples were identified to be hub genes among the top 10 genes. In vitro experiments were performed to investigate the influence of TRP extracts on the cell metabolic activity, migration, invasion, apoptosis, and proliferation of two OSCC cell lines (CAL-27 and SCC-9). The functional rescue assay was conducted to investigate the effect of applying the inhibitor and activator of an enriched pathway on the phenotypes of cancer cells. In addition, the zebrafish xenograft tumor model was established to investigate the influence of TRP extracts on tumor growth and metastasis in vivo. RESULTS: The target genes of TRP in OSCC were prominently enriched in the PI3K-Akt signaling pathway, with the identification of five hub genes (JUN, EGFR, ESR1, RELA, and AKT1). TRP extracts significantly inhibited cell metabolic activity, migration, invasion, and proliferation and promoted cell apoptosis in OSCC cells. Notably, the application of TRP extracts exhibited the capacity to downregulate mRNA and phosphorylated protein levels of AKT1 and ESR1, while concomitantly inducing upregulation of mRNA and phosphorylated protein levels in the remaining three hub genes (EGFR, JUN, and RELA). The functional rescue assay demonstrated that the co-administration of TRP and the PI3K activator 740Y-P effectively reversed the impact of TRP on the phenotypes of OSCC cells. Conversely, the combination of TRP and the PI3K inhibitor LY294002 further enhanced the effect of TRP on the phenotypes of OSCC cells. Remarkably, treatment with TRP in zebrafish xenograft models demonstrated a significant reduction in both tumor growth and metastatic spread. CONCLUSIONS: Triphala exerted significant inhibitory effects on cell metabolic activity, migration, invasion, and proliferation in OSCC cell lines, accompanied by the induction of apoptosis, which was mediated through the inactivation of the PI3K/Akt pathway.

Quantum dot imaging for HSP70 and HSF­1 kinetics in SCC­25 cells with or without leucine deprivation following heat shock.

The aim of this study was to develop a quantum dot-based approach for heat shock protein 70 (HSP70) and heat shock factor 1 (HSF-1) kinetics following heat shock, and to discover approaches to thermotherapy based on disrupting the effect of activation of HSF-1 and the accumulation of HSP70 by leucine deprivation. SCC-25 cells cultured with limiting leucine or normal leucine were stressed at 42˚C for 30 min, and were cultured for 2, 4, 6, 8 and 10 h, respectively. The expression of HSP70 and HSF-1 was observed using confocal laser microscopy and semi-quantitative analysis was performed by Image-Pro Plus. At 6 h after heating, HSF-1 in cells cultured with normal leucine was activated and translocated from the cytosol to the nucleus, and the synthesis of HSP70 reached the maximum value and had a tendency to gather in the nucleus. However, in cells cultured with limiting leucine, HSF-1 activity decreased and accumulation of HSP70 was not found. Leucine deprivation results in the inactivation of HSF-1 leading to slight accumulation of HSP70 and no tendency to gather in the nucleus. Thus, HSF-1 may serve as a novel therapeutic target in the treatment of oral cancer.