| Objective: To investigate the molecular mechanism of coenzyme Q10B (COQ10B) affecting the malignant biological behavior of esophageal squamous cell carcinoma (ESCC) cells. Methods: qRT-PCR was used to detect the expression of COQ10B in 3 human ESCC cell lines (KYSE150, KYSE450 and TE-1) and esophageal epithelial cell line Het-1A. To knock on reducing COQ10B KYSE150 cells RNA sequencing screening differentially expressed genes, through KEGG pathway analysis for closely related signaling pathway. KYSE150 and TE-1 cells stably overexpressing COQ10B were constructed by lentivirus, and the overexpression efficiency was detected by Western blot. CCK8 method is used to detect the PI3K/AKT signal pathway inhibitor LY294002 for esophageal squamous cancer cells half inhibitory concentration (IC50) values. Through EDU, plate cloning experiment, flow cytometry, wound healing, Transwell invasion chamber express COQ10B and tested after joining LY294002 inhibitors of esophageal squamous cancer cell proliferation, apoptosis, migration and invasion ability of influence. Western blot was used to detect the effects of COQ10B overexpression and LY294002 inhibitor on the expression of PI3K/AKT signaling pathway related proteins (PI3K, AKT, p-PI3K and p-AKT) in KYSE150 esophageal squamous cell carcinoma cells. Results: COQ10B was relatively highly expressed in ESCC cell lines. Based on RNA transcriptome sequencing technology, a total of 319 significantly differentially expressed genes were screened. Cut genes which raised 285 genes, 34, through KEGG pathway analysis selecting and PI3K/AKT signaling pathway as the research object of subsequent molecular mechanisms. LY294002, an inhibitor of PI3K/AKT signaling pathway, showed an enhanced toxic effect on ESCC cells with the increase of concentration and action time. In the subsequent experiments, the IC50 value of LY294002 for 48 hours was used. Compared with the negative control group, overexpression of COQ10B in KYSE150 and TE-1 cells significantly enhanced cell proliferation, migration and invasion and inhibited cell apoptosis. However, after treatment with LY294002 inhibitor for 48h, the proliferation, migration and invasion abilities of ESCC cells enhanced by COQ10B overexpression were significantly inhibited, while the inhibition of apoptosis ability was reversed. Compared with the negative control group, the expression levels of p-PI3K and p-AKT in PI3K/AKT signaling pathway were significantly increased after overexpression of COQ10B in KYSE150 esophageal squamous cell carcinoma cells. Treatment with LY294002 inhibitor for 48 hours significantly inhibited the expression of p-PI3K and p-AKT, which were related proteins in PI3K/AKT signaling pathway enhanced by COQ10B overexpression. Conclusion: COQ10B can through the activation of PI3K/AKT signaling pathway to promote esophageal squamous cancer cell proliferation, migration, invasion, and inhibiting apoptosis. |
