Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025056
Triptonide (TN) is a small-molecule compound initially derived from Tripterygium wilfordii Hook. f used in traditional Chinese medicine. However, its potential antitumor mechanisms are still far from adequately understood. The purpose of this research is to elucidate the antitumor and pharmacological effects of TN on esophageal squamous cell carcinoma (ESCC). Functional assays, such as CCK-8 and colony formation assays, are used to evaluate the effects of TN on KYSE450 and KYSE510 cells. Subsequently, western blot analysis, Hoechst 33258 staining, flow cytometric analysis, autophagic flux detection, and transmission electron microscopy (TEM) are used to determine the effects of TN on apoptosis and autophagy in ESCC cells. Additionally, the autophagy inhibitor 3-methyladenine (3-MA) and the AMPK inhibitor dorsomorphin (Compound C, CC) are administered to explore the molecular mechanisms and crucial pathways in ESCC cells. Our findings provide strong evidence that TN induces autophagy-dependent apoptosis by targeting the AMPK-mTOR-ULK1 axis in ESCC cells. Collectively, this study sheds light on the anticancer mechanisms of TN in esophageal squamous cell carcinoma and suggests that TN is a promising candidate for the antitumor phytomedicine.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21331
BACKGROUND: Age-related degeneration is closely associated with bone metabolic imbalance. In the jaw, this manifests as alveolar bone resorption, tooth loosening, and even loss. Impaired osteogenic differentiation potential of senescent jaw bone marrow mesenchymal stem cells is a critical factor hindering jaw bone regeneration. Quercetin, a natural flavonoid compound, exhibits antioxidant, anti-inflammatory, and cell differentiation-regulating properties, yet effect and mechanism of quercetin in osteogenic differentiation of senescent jaw bone marrow mesenchymal stem cells remain unclear. OBJECTIVE: To investigate the effects of quercetin on the proliferation, migration, osteogenic differentiation, and senescence of aged jaw bone marrow mesenchymal stem cells. METHODS: Jaw bone marrow mesenchymal stem cells were isolated from the mandibles of 10 8-week-old SD rats and cultured using a combination of bone marrow flushing and bone slice digestion. Jaw bone marrow mesenchymal stem cells were subcultured to the third and seventh passages, serving as the young and senescent groups, respectively. The quercetin group was treated with quercetin based on the senescent group. CCK-8 assay was used to detect the effects of 0.01, 0.1, 1, 10, 100 μmol/L quercetin on proliferation of senescent jaw bone marrow mesenchymal stem cells, and the optimal concentration was selected. Cell scratch assay was used to observe cell migration ability. RT-qPCR and western blot were used to detect the expression of senescence markers. β-galactosidase staining was used to observe the proportion of positive cells. After 7 days of osteogenic induction, RT-qPCR and western blot were used to detect the expression of osteogenic markers. After 14 days of osteogenic induction, alkaline phosphatase staining was performed. After 21 days of osteogenic induction, alizarin red staining was performed. Western blot was used to detect the expression of phosphorylated protein kinase B, protein kinase B, phosphorylated mammalian target of rapamycin, and mammalian target of rapamycin. RESULTS AND CONCLUSION: Compared with the young group, the proliferation ability of the senescent group decreased. Compared with the senescent group, 1 μmol/L quercetin significantly promoted the proliferation of senescent jaw bone marrow mesenchymal stem cells (P < 0.01). Compared with the senescent group, the migration ability of senescent jaw bone marrow mesenchymal stem cells in the quercetin group was improved, the proportion of β-galactosidase positive cells was significantly reduced, and the mRNA and protein expression of senescence-related P16, P53, and P21 were decreased (P < 0.05). After osteogenic induction, compared with the senescent group, the quercetin group showed increased calcium nodule formation, alkaline phosphatase staining area, and mRNA and protein expression of alkaline phosphatase, osteopontin, and Runt-related transcription factor 2 (P < 0.05). Compared with the senescent group, the phosphorylation levels of protein kinase B and mammalian target of rapamycin in the quercetin group were significantly reduced (P < 0.05). These results indicate that quercetin can inhibit senescence of jaw bone marrow mesenchymal stem cells caused by multiple passages and promote osteogenic differentiation by regulating the protein kinase B/mammalian target of rapamycin signaling pathway.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025056
Esophageal squamous cell carcinoma (ESCC) accounts for 90% of global esophageal cancer cases, with high prevalence in Asia, East Africa, and South America. Current treatment relies on esophagectomy, which carries significant morbidity and limited efficacy, underscoring the urgent need for novel therapeutic agents. Triptonide (TN), a small-molecule compound derived from Tripterygium wilfordii Hook. f, exhibits potent antitumor activity, but its mechanisms in ESCC remain undefined. This study investigated TN-induced cytotoxicity in KYSE450 and KYSE510 ESCC cell lines using CCK-8 and colony formation assays. Western blot analysis, Hoechst 33258 staining, flow cytometry, autophagic flux detection, and transmission electron microscopy (TEM) assessed apoptosis and autophagy. Pharmacological inhibitors 3-methyladenine (3-MA) and dorsomorphin (Compound C, CC) were employed to dissect pathway involvement. Results demonstrate that TN induces autophagy-dependent apoptosis by targeting the AMPK-mTOR-ULK1 axis. TN treatment led to AMPK activation, mTOR inhibition, and ULK1 phosphorylation, triggering autophagic flux and subsequent apoptosis. Inhibition of autophagy with 3-MA or AMPK with CC attenuated TN-induced apoptosis, confirming the mechanistic dependency. These findings establish TN as a promising candidate for ESCC therapy and elucidate a novel molecular pathway for its anticancer effects.