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Objective To investigate the impacts of forkhead box M1(FOXM1)on the proliferation,invasion,and drug resistance of gastric cancer cells by regulating the circular RNA circ_NOTCH1.Methods Western blotting and real-time quantitative PCR were performed to determine the expression of FOXM1 protein and circ_NOTCH1,respectively,in the gastric cancer tissue,para-carcinoma tissue,human normal gastric mucosa epithelial cell line GES-1 and gastric cancer cell lines MGC-803,HGC-27,and BGC-823.BGC-823 cells were classified into the following groups:control,short hairpin RNA FOXM1(sh-FOXM1)and negative control(sh-NC),small interfering RNA circ_NOTCH1(si-circ_NOTCH1)and negative control(si-NC),and sh-FOXM1+circ_NOTCH1 overexpression plasmid(sh-FOXM1+pcDNA-circ_NOTCH1)and sh-FOXM1+negative control(sh-FOXM1+pcDNA).CCK-8 assay and clone formation assay were employed to measure the cell proliferation,and Transwell assay to measure cell invasion.After treatment with 1.0 mg/L adriamycin for 48 h,the cell resistance in each group was analyzed.Western blotting was employed to determine the expression levels of FOXM1,proliferating cell nuclear antigen(PCNA),Bax,multi-drug resistance-associated protein 1(MRP1),and multi-drug resistance gene 1(MDR1).RNA pull-down and RNA immunoprecipitation were employed to examine the binding of circ_NOTCH1 to FOXM1 protein.Results Compared with those in the para-carcinoma tissue,the expression levels of FOXM1 protein and circ_NOTCH1 in the gastric cancer tissue were up-regulated(all P<0.001).Compared with GES-1 cells,MGC-803,HGC-27,and BGC-823 cells showed up-regulated expression levels of FOXM1 protein and circ_NOTCH1(all P<0.001).Compared with the control group and sh-NC group,the sh-FOXM1 group with down-regulated expression of FOXM1 protein and circ_NOTCH1 showed decreased optical density value,clone formation rate,cell invasion number,and cell viability,down-regulated expression of PCNA,MRP1,and MDR1,and up-regulated expression of Bax protein in BGC-823 cells(all P<0.001).Compared with the control group and the si-NC group,the si-circ_NOTCH1 group with down-regulated expression of circ_NOTCH1 showed decreased optical density value,clone formation rate,cell invasion number,and cell viability,down-regulated expression of PCNA,MRP1,and MDR1,and up-regulated expression of Bax protein in BGC-823 cells(all P<0.001).Compared with sh-FOXM1 group and sh-FOXM1+pcDNA group,the sh-FOXM1+pcDNA-circ_NOTCH1 group with up-regulated expression of circ_NOTCH1 showed increased optical density value,clone formation rate,cell invasion number,and cell viability,up-regulated expression of PCNA,MRP1,and MDR1,and down-regulated expression of Bax protein(all P<0.001).FOXM1 protein was able to interact with circ_NOTCH1.Conclusion Interference with FOXM1 may inhibit the proliferation,invasion,and drug resistance of gastric cancer cells by silencing circ_NOTCH1 expression.
Subject(s)
Humans , bcl-2-Associated X Protein/metabolism , Carcinoma , Cell Line, Tumor , Cell Proliferation/genetics , Drug Resistance , Forkhead Box Protein M1/metabolism , Gene Expression Regulation, Neoplastic , MicroRNAs/genetics , Proliferating Cell Nuclear Antigen/metabolism , Receptor, Notch1/metabolism , RNA, Small Interfering/genetics , Stomach Neoplasms/geneticsABSTRACT
Objective@#To check Forkhead box M1 (FoxM1) expression in nasal mucosal of chronic rhinosinusitis (CRS) patients and the effect of inflammatory factors on FoxM1 expression, in order to research the significance of FoxM1 in CRS.@*Methods@#From January to October of 2018, 50 patients hospitalized in the Department of Otorhinolaryngology, Head and Neck Surgery, First Affiliated Hospital of Nanchang University were enrolled in this study. Twenty CRS patients with polyps (CRSwNP), 20 CRS patients without nasal polyps (CRSsNP) and 10 patients with simple deviation of nasal septum (the control groups) were selected. The expression of FoxM1 in nasal mucosa of these patients was detected by immunohistochemistry (IHC) and real-time fluorescent quantitative PCR (qRT-PCR). Meanwhile, HE stain was used to observe the pathologic changes in each sample. By establishing human nasal epithelium cells cultivating model in vitro and identifying via immumofluorescence method, experimental group and control group were set up, then activation factors including interleukin (IL)-1β, IL-4, IL-5, IL-17, interferon-γ (IFN-γ) and staphylococcal entemtoxin B (SEB) were added in the models after stabilizing passage, and qRT-PRC and Western blot method were applied to check the expressing change of FoxM1. Software SPSS 18.0 was used for statistical analysis.@*Results@#HE stain showed that the mainly pathologic change in nasal mucosa of CRS patients with or without nasal polyp was mucosal epithelial cells, goblet cell and submucosal gland hyperplasia, accompanied by a large number of inflammatory cells infiltration. The result of IHC demonstrated that both of the expression of FoxM1 in nasal mucosal tissue of CRS patients in the CRSwNP and CRSsNP groups exceed that of the control group (80% vs 75% vs 20%, χ2 value was 10.000, 8.213, respectively, all P<0.05); there was no difference of expression between the two groups of CRS patients (χ2=0.143, P>0.05). The result of qRT-PCR demonstrated that the expression of FoxM1 mRNA in nasal mucosa of CRSwNP and CRSsNP was increased compared with that of the control group (3.309±1.511 vs 3.261±1.336 vs 1.000±0.774, t value was 4.519, 4.928, respectively, all P<0.05), but the difference between the two groups of CRS patients had no statistic significance (t=0.107, P=0.909). Nasal mucosa epithelial cells cultivating models was established successfully. Q-RT PCR and Western blot were conducted after stimulation of 100 ng/ml IL-1β, IL-4, IL-5, IL-17, IFN-γ and SEB for 36 h, and the proteins expression levels of FoxM1 exceeded the groups without stimulation with statistic significance.@*Conclusions@#The expression of FoxM1 in CRS increases and many types of cytokine can induce the increase of FoxM1 in human nasal epithelial cells. FoxM1 may participate in the process of pathogenesis in CRS.
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Objective: To investigate the effects of silencing the expressionof Forkhead box M1 (FOXM1) gene by the specific siRNA on proliferation, apoptosis and chemosensitivity of human nasopharyngeal carcinoma cells, and to explore the molecular mechanisms. Methods: The specific siRNA fragments targeting FOXM1 gene (FOXM1-siRNA) was transfected into nasopharyngeal carcinoma 5-8F cells, then the silencing efficiency of FOXM1 gene expression was detected by RT-PCR, real-time fluorescent quantitative PCR and Western blotting, respectively. The changes of proliferation ability, cell cycle distribution, apoptosis rate, and paclitaxel sensitivity of 5-8F cells after FOXM1-siRNA transfection were detected by MTT assay, FCM, and AnnexinV-FITC/PI staining assay, respectively. The expressions of relative proteins in 5-8F cells with FOXM1 gene silencing were detected by Western blotting. Results: The expressions of FOXM1 mRNA and protein in 5-8F cells after FOXM1-siRNA transfection were significantly decreased (both P < 0.01). After FOXM1 gene silencing, the proliferation ability of 5-8F cells was decreased (P < 0.05), and the expression level of proliferating cell nuclear antigen (PCNA) protein was significantly decreased (P < 0.01). Meanwhile the proportion of cells in G1 phase after FOXM1 gene silencing was increased (P < 0.01), while the proportion of cells in S phase was decreased (P < 0.05), and the expression of Cyclin D1 was down-regulated (P < 0.01). The apoptosis rate of 5-8F cells with FOXM1 gene silencing was significantly increased (P < 0.01), the expression level of Bcl-2 was down-regulated (P < 0.01), and the expression level of Bax was up-regulated (P < 0.01). Furthermore, the sensitivity of 5-8F cells to paclitaxel was significantly enhanced (P < 0.01), and the expression of multidrug resistance-associated protein 1 (MRP1) was down-regulated (P < 0.01) after FOXM1-siRNA transfection. Conclusion: The specific siRNA silencing FOXM1 gene expression can effectively inhibit the proliferation of nasopharyngeal carcinoma 5-8F cells, promote cell apoptosis, and enhance the sensitivity to paclitaxel. These effects may be related to the down-regulation of PCNA, Cyclin D1, MRP1 and Bcl-2 expressions as well as the up-regulation of Bax expression in 5-8F cells.
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The proliferation, progression and metastasis of colorectal cancer is a pathological process involving many factors. Human epidermal growth factor receptor 2 (HER2) and forkhead box M1 (FOXM1) are two oncogenes, and their encoded proteins play important roles in cell metabolism, proliferation, apoptosis, migration and invasion processes. FOXM1 may be a downstream effector regulated by HER2. Both of HER2 and FOXM1 are highly expressed in colorectal cancer, and correlated with clinicopathological parameters and prognosis. Therefore, HER2 and FOXM1 maybe have important reference value in the diagnosis, treatment and prognosis of colorectal cancer, and may be new therapeutic targets for colorectal cancer. This paper reviews the mechanisms of HER2 and FOXM1 action in colorectal cancer, the clinical significance of their expressions, and the correlation between the two proteins.
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AIM: To investigate the role of Forkhead box M 1 ( FoxM1 ) and B-cell leukemia/lymphoma-2 (Bcl-2) in the pathogenesis of acute myeloid leukemia (AML).METHODS:RT-qPCR and immunofluorescence analysis were used to determine the expression of FoxM 1 at mRNA and protein levels in AML-de novo patients, AML-complete re-mission (CR) patients, AML-refractoriness and relapse (RR) patients and healthy controls.HL60 cells and K562 cells were transfected with FoxM1 siRNA.The cell proliferation was detected by cell proliferation assay and colony formation as-say on soft agar, and the cell apoptosis was determined by flow cytometry .The expression of FoxM1 and Bcl-2 at mRNA and protein levels was detected by RT-qPCR and Western blotting .The activity of bcl-2 promoter was examined by lucifer-ase reporter assay with FoxM1 targetting.RESULTS:FoxM1 expression level in the AML-de novo patients was significant-ly higher than that in the healthy controls .As compared with the AML-de novo patients, FoxM1 expression in the AML-CR patients was reduced , and the FoxM1 expression level was the highest in the AML-RR patients .FoxM1 expression was in-hibited in the HL60 cells and K562 cells transfected with FoxM1 siRNA.Transfection with FoxM1 siRNA in the HL60 cells and K562 cells inhibited the proliferation as compared with NC siRNA transfection , and impaired the colony formation abili-ty.On the contrary , transfection with FoxM1 siRNA promoted the cell apoptosis .FoxM1 regulated bcl-2 expression posi-tively.CONCLUSION:FoxM1 promotes the development of AML by regulating bcl-2 expression.Silencing of FoxM1 ex-pression suppresses cell proliferation and promotes cell apoptosis .FoxM1 is a potential target for AML treatment .
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FOXM1 (Forkhead box M1), one factor of the Forkhead family, has three subtypes (FOXM1a, FOXM1b, FOXM1c). The current study focuses on FOXM1b and FOXM1c. FOXM1 regulates transcription of prolifemtion-nssociated genes and plays a vital role in embryogenesis as well as reorgani-zation. Recent studies have shown that FOXM1 is closely related with tumor occurrence and development. Tumors with high expression of FOXM1 are often poorly differentiated, highly malignant, distantly metastat-ic, poorly predicted. FOXM1 has a influence on the tumor proliferation, invasion, metastasis and angiogene-sis through regulating its downstream tumor-related genes. At present, the synthesis and study of the anti-tumor chemicals targeting FOXM1 offer a possibility of the FOXM1 in clinical applications. More and more researchers attach importance to its tumor therapeutic value both at home and aboard. This paper will make a review about the lastest FOXM1 research in oncology.