MCTS1 Re-Initiation and Release Factor (MCTS1) has been characterised as an oncoprotein in some cancers. In this study, we explored the expression of MCTS1 in laryngeal squamous cell carcinoma (LSCC) and its regulatory effects on the proliferation and cell-cycle progression of tumour cells, as well as the underlying mechanisms.The data from the Cancer Genome Atlas was used to analyse MCTS1 expression and its correlation with survival outcomes in LSCC patients. Subsequent in vitro cellular and molecular studies were performed based on representative LSCC cell lines.Results showed that the upregulation of MCTS1 in LSCC is linked to poor progression-free survival (PFS) and disease-specific survival (DSS). In TU177 and AMC-HN-8 cells, MCTS1 exerted positive regulations on cell viability, colony formation, cell cycle progression, and the expression of CDK1, CDK2, cyclin A2, and cyclin B1. Co-IP assay confirmed mutual interaction between MCTS1 and LARP7, mainly in the cytoplasm. Cycloheximide (CHX) chase and co-IP assay of ubiquitination showed that MCTS1 could increase LARP7 protein half-life and reduce its poly-ubiquitination.LARP7 overexpression enhanced the viability and colony formation of LSCC cells and also elevated the expression of CDK1, CDK2, cyclin A2, and cyclin B1. In addition, its overexpression partly reversed the negative influence of MCTS1 knockdown. In summary, this study confirmed that the expression of MCTS1 might be an indicator of unfavourable prognosis for patients with LSCC. Mechanically, it promotes LSCC cell viability and proliferation via interacting with LARP7 and reducing its proteasomalmediated degradation.
Objectives: Study on the chemotherapeutic effect and mechanism of cucurbitacin E (CuE) on laryngeal cancer stem cells. Methods: We used flow cytometry to sort out CD133 + laryngeal cancer stem cells; trypan blue rejection assay to detect the survival rate of laryngeal cancer stem cells; Cell counting kit-8 (CCK-8) assay to detect the effect of CuE on the proliferation ability of stem cells and the chemotherapeutic potentiation of doxorubicin; Transwell assay to observe the effect of CuE on the migration ability of stem cells; and Western Blot to detect the effect of CuE on the expression level of stem cell-associated proteins. The tumor volume of nude mice was measured at the end of the experiment, and paraffin sections of nude mice tumor tissues were prepared and stained with Hematoxylin and eosin (H&E). The expression of c-MYC in tumor tissues of nude mice was further detected by immunohistochemistry, and the effect of CuE on the expression level of related proteins in tumor tissues of nude mice was detected by Western Blot.Results: CuE reduced the survival rate, proliferation ability, and migration ability of laryngeal cancer stem cells in vitro, and that CuE had a chemotherapeutic potentiating effect on doxorubicin. The possible mechanism of the chemotherapeutic effect of CuE was to reduce the expression of c-MYC protein, and the possible mechanism of chemotherapy synergy was to reduce the expression of ABCG2 and P-gp protein. Conclusion:CuE has a chemotherapeutic effect on laryngeal cancer stem cells, as well as a chemotherapy synergy.
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