Background Circular RNA hsa_circ_0003340 (circ-OGDH) has been uncovered to be involved in esophageal squamous cell carcinoma (ESCC) progression. However, the mechanism by which circ-OGDH regulates ESCC progression is unclear. Methods Expression levels of circ-OGDH, microRNA (miR)-615-5p, and PDX1 (pancreatic and duodenal homeobox 1) mRNA were evaluated with quantitative real-time polymerase chain reaction (qRT-PCR). The proliferation, apoptosis, migration, invasion, and cell cycle progression of ESCC cells were analyzed by MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide), colony formation, flow cytometry, and transwell assays. Measurement of glutamine consumption, α-KG (α-ketoglutarate) production, and ATP (Adenosine Triphosphate) content using corresponding kits. Protein levels were analyzed by Western blotting. The targeting relationship between circ-OGDH or PDX1 and miR-615-5p was verified by dual-luciferase reporter and RNA immunoprecipitation (RIP) assays. The function of circ-OGDH in ESCC was confirmed by animal experiments. Results Circ-OGDH was upregulated in ESCC. Circ-OGDH inhibition reduced ESCC growth in vivo and accelerated cell apoptosis, cell cycle arrest, repressed cell proliferation, migration, invasion, and reduced cell glutamine metabolism in ESCC cells in vitro. MiR-615-5p was downregulated in ESCC, while PDX1 had an opposite result. Circ-OGDH sponged miR-615-5p to regulate PDX1 expression. MiR-615-5p inhibitor neutralized the repressive effect of circ-OGDH knockdown on malignancy and glutamine metabolism of ESCC cells. PDX1 overexpression counteracted the inhibitory impact of miR-615-5p mimic on malignancy and glutamine metabolism of ESCC cells. Conclusion Circ-OGDH sponged miR-615-5p to elevate PDX1 expression, thus elevating glutamine metabolism and promoting tumor growth in ESCC. The study offered evidence to support circ-OGDH as a promising target for ESCC therapy.
Lung cancer is a leading cause of cancer-associated mortality and morbidity worldwide. Previous studies have suggested that ATP-binding cassette transporter E1 (ABCE1) acetylation is upregulated in the tissues and cells of lung cancer and is associated with the prognosis of patients with lung cancer. The aim of the present study was to investigate the association between Tat interactive protein 60 kDa (Tip60) expression and ABCE1 acetylation, and the effect of Tip60 on the biological functions of A549 lung carcinoma cells. The expression levels of Tip60 and ABCE1 acetylation were examined using western blot and co-immunoprecipitation (Co-IP) assays in normal bronchial epithelial (HBE) and human lung cancer (A549) cells. The expression of Tip60 then was downregulated in A549 cells using small interfering RNA. Wound healing and Transwell assays were used to assess cell invasion and migration. The biological effects of Tip60 in lung cancer cells were investigated using MTT and flow cytometric assays. Subsequently, tumor xenografts were established to observe the effect of Tip60 on lung cancer in vivo. Western blot and Co-IP assays were performed to investigate the mechanism of Tip60 in A549 cells. Tip60 expression and ABCE1 acetylation were upregulated in the lung cancer cells compared with the normal bronchial epithelial cells. Downregulation of Tip60 decreased the acetylation of ABCE1 and inhibited cell proliferation, invasion and migration. Furthermore, the downregulation of Tip60 activated the apoptotic pathway in order to achieve its suppressive function. In the xenografts, the tumor weight and volume were notably reduced due to the downregulation of Tip60 expression. The results of the present study strongly suggest that Tip60 is a novel target in the prevention and treatment of lung cancer.
Background: Esophageal cancer is one of the primary death causes leading by cancer in the world, which is high morbidity and mortality. Epigenetic acetylation modification participates in and regulates the proliferation, invasion, and metastasis of various tumor cells, and the acetylation modification of tumor proteins involved by acetyltransferases may be one of the important mechanisms of esophageal carcinogenesis. The aim of this study was to investigate the correlation of acetyltransferase P300 and Survivin acetylation in esophageal cancer pathogenesis and its molecular mechanism. Methods: Fifty-five cases of esophageal cancer tissues and adjacent cancer tissues were collected, Survivin and P300 protein expression was measured by immunohistochemistry (SP) and protein blotting (Western Blot); Survivin acetylated protein levels were measured by coimmunoprecipitation (Co-IP); bioinformatics predicted the relationship between P300 and Survivin as the substrate, and fluorescence immunohistochemistry (IF) to verify the localization and expression of Survivin and P300 in esophageal cancer tissues; the correlation of Survivin acetylation, P300 and clinical cases characteristics was analyzed by statistics. P300 siRNA sequences were structured and transfected into EC109 cells. P300 protein expression and Survivin acetylated protein levels were determined by Co-IP. Cell viability was determined by the MTT assay, Scratch healing and Transwell chamber assay examined cell migration and invasion ability. Results: Survivin and P300 protein expression was significantly increased in human esophageal cancer tissues and EC109 cells. The Survivin protein was acetylated in esophageal cancer tissues and EC109 cells, and its protein acetylation rate was significantly increased; bioinformatics predicted that the acetyltransferase P300 could catalyze the acetylation of Survivin as a substrate, and the fluorescence immunohistochemistry confirmed that both Survivin and P300 simultaneously showed a high expression state in cancer
Objective: To investigate the effect of Tip60 gene silencing on the ABCE1 acetylation level and cell proliferation, migration and invasion in TE-1 cells of oesophageal cancer. Methods: The siRNA sequence of Tip60 was transfected with esophageal cancer TE-1 cells. Transfected siRNA vector cells were used as experimental group (si-T), siRNA no-loaded somatic cells were transfected as control group (si-NC), and untransfected TE-1 cells were used as blank group (Group N). ABCE1 mRNA was detected by qRT-PCR, the expression of ABCE1 protein, proliferation-related protein β catenin (β-catenin), GSK3β, and c-myc by Western blot, the protein acetylation level by immunoprecipitation, MTT assay for cell viability, scratch healing and Transwell compartment assay for migration and invasion ability. Results: After 48 h downregulation of the Tip60 gene, TE-1 cells showed no significant changes in the ABCE1 mRNA and protein expression. The acetylation level of ABCE1 decreased significantly, compared with the control group and the blank group.After Tip60 gene silencing, the expression of β-catenin and c-myc protein decreased, while the expression of GSK-3β protein increased. Cytofunctology experiments showed that the proliferative activity, migration and invasion ability of TE-1 cells in the experimental group were significantly inhibited. Conclusion: Down regulation of Tip60 gene can deacetylate ABCE1 protein and inhibit the proliferation activity, migration and invasion ability of esophageal cancer by blocking the conduction of Wnt signaling pathway.
Objective: The incidence of primary lung adenocarcinoma is increasing year by year in worldwide, and it has surpassed squamous cell carcinoma to become the most common pathological type of lung cancer. Advances in medical technology have made the overall survival rate of lung cancer patients improve, but it is still very low. In the study, we investigated the methyltransferase SETDB1 expression and the methylation level of SPG20, and analyzed the correlation and its clinical significance in the pathogenesis of lung adenocarcinoma. Methods: 60 lung adenocarcinoma and normal lung tissues were selected.The expression of SPG20 and SETDB1 mRNA was examined by RT-qPCR; Protein expression of SPG20 and SETDB1 was determined by immunohistochemistry(SP) and immune protein imprinting(Western blot);Methylation level of SPG20 gene was determined by pyrosequencing. Correlation between SETDB1 and SPG20 methylation levels and both and clinical case characteristics was analysed by statistics. Western blot detection of SETDB1 and SPG20 protein expression in lung adenocarcinoma cells and normal bronchial mucosal epithelial cells.The small interfering RNA of SETDB1 was transfected into lung adenocarcinoma A549 cells by liposome-mediated method, the mRNA and protein expression levels of SETDB1 were detected by RT-qPCR and Wesrern blot, and the methylation level of SPG20 gene was detected by pyrosequencing. Results:Relative expression of SPG20 mRNA was significantly lower in lung adenocarcinoma tissue than in normal lung tissue, while relative SETDB1 mRNA expression was higher in cancer tissue than in normal lung tissue. SETDB1 protein expression was significantly higher in lung adenocarcinoma tissue than in normal lung tissue, while SPG20 protein showed lower expression in cancer tissue than in normal lung tissue. The methylation rate of the SPG20 gene was significantly higher in cancerous tissues than in normal lung tissue, the difference was statistically significant. SPG20 methylation in cancer tissues showed a correlation with SETDB1 and showed a positive correlation; SPG20 methylation and SETDB1 are closely related with TNM stage, tissue differentiation, and lymph node metastasis in lung adenocarcinoma. The vitro experiments showed that SETDB1 was highly expressed, while SPG20 was lowIn lung adenocarcinoma cells, SETDB1 was low and SPG20 was high in normal bronchial epithelial cells. RNA interference with SETDB1 could significantly reduce the mRNA and protein expression of SETDB1 in A549 cells, the methylation rate of SPG20 gene was significantly decreased, and the protein expression was increased. Conclusion:There is a significant correlation between methyltransferases SETDB1 and SPG20 methylation in lung adenocarcinoma, and high SETDB1 expression may be the upstream molecule of SPG20 methylation, working together in promoting the occurrence and development of lung adenocarcinoma.
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