| Objective: To investigate the effect of 3-BrPA on proliferation,migration and invasion of human Hep2 laryngeal carcinoma cells by culturing human Hep2 laryngeal carcinoma cells in vitro,and to explore the possible mechanism of 3-Br PA based on PI3K/AKT/m TOR signaling pathway,providing experimental basis for early diagnosis and precise treatment of laryngeal carcinoma.Methods: Screening for differential proteins in laryngeal cancer with glycolytic profiles based on the TCGA database and proteins associated with the glycolytic pathway.STRING database was used to analyze the correlation between PI3 K,AKT,m TOR and HK-2 and TGFBI proteins.Human Hep2 laryngeal carcinoma cells were cultured in vitro and divided into control,3-BrPA 95,115,and 135 μmol/L groups.CCK-8 assay was applied to examine survival rate of Hep2 laryngeal carcinoma cells,and the morphological variations of Hep2 laryngeal carcinoma cells were observed by inverted phase contrast microscope.The effect of3-Br PA on the migration of Hep2 laryngeal carcinoma cells was observed by scratch assay,and the effect of 3-Br PA on the invasion of Hep2 laryngeal carcinoma cells was observed by Transwell assay.The expression of PI3 K,AKT,m TOR,HK-2 and TGFBI m RNA and protein were detected by RT-q PCR and Western bolt assay,respectively.Results: Bioinformatics analysis showed that TGFBI was highly expressed in laryngeal carcinoma tissues with glycolytic metabolism characteristics.STRING database showed that AKT,m TOR and HK-2 were highly correlated.The results of CCK-8 experiment showed that the proliferation viability of laryngeal cancer cells was significantly decreased in the experimental group compared with the control group,and the difference was statistically significant(p<0.05,p<0.01).3-Br PA effect on Hep2 laryngeal carcinoma After 24 h,the cell morphology changed significantly: compared with the control group,with the increase of drug concentration,the cells gradually became smaller,rounded and shrunken,the cell gap gradually widened,vacuoles were visible in the cytoplasm,and the number of dead suspended cells gradually increased.Transwell invasion assay showed that the number of Hep2 laryngeal cancer cells invading at 24 h decreased gradually with the increase of drug concentration,and the decrease of migration rate was statistically significant compared with the control group(p<0.01).RT-q PCR results showed that the expression of PI3 K,AKT,m TOR,HK-2 and TGFBI m RNA decreased in each experimental group compared with the control group after3-Br PA treatment of laryngeal cancer cells for 24h(p<0.05,p<0.01).Western bolt results showed that the expression of PI3 K,AKT,p-AKT and m TOR,HK-2 and TGFBI proteins were significantly reduced in each experimental group after 3-Br PA treatment of laryngeal cancer cells for 24 h compared with the control group(p<0.01).Conclusion: 1.3-BrPA inhibited the proliferation,migration and invasion of human Hep2 laryngeal carcinoma cells;2.The mechanism of action of 3-Br PA to inhibit the proliferation,migration and invasion of human Hep2 laryngeal cancer cells may be related to the regulation of PI3K/AKT/m TOR signaling pathway,down-regulation of HK-2 and TGFBI expression,and inhibition of glycolysis. |
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