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Objective To explore the effect of FBXW7 on ferroptosis in head and neck squamous cell carcinoma. Methods Head and neck squamous cell lines HN4 and HN6 were cultured in vitro. FBXW7 and SOX2 overexpression plasmids were constructed, and the plasmids were stably transfected into cell lines. The overexpression transfection efficiency was verified at the transcription level and protein level by qRT-PCR and Western blot experiments, respectively. The lipid peroxidation levels of head and neck squamous cell carcinoma cells with overexpressing FBXW7 were verified by measuring malondialdehyde(MDA), glutathione(GSH), and reactive oxygen species(ROS) levels. After treating cells with ferroptosis inhibitor Fer-1, the changes in cell viability were further detected to verify the effect of FBXW7 on ferroptosis. The effect of transfection of the overexpressed plasmid on cellular pathways was detected by Western blot. Results HN4 and HN6 cell lines showed increased levels of lipid peroxidation after overexpression of FBXW7, and the ferroptosis inhibitor Fer-1 was able to effectively reverse the ferroptosis induced by overexpression of FBXW7. Western blot assay results showed that overexpression of FBXW7 reduced the expression of c-Myc, SOX2 and SLC7A11. Conclusion FBXW7 regulates the expression of SOX2-SLC7A11 by degrading c-Myc, thereby effectively regulating ferroptosis in HNSCC.

Objective: Our study aimed to probe the potential molecular mechanism of long non ⁃ coding RNA (lncRNA) VIM Antisense RNA 1 (VIM⁃AS1) in diabetic retinopathy. Methods: LncRNA VIM⁃AS1, miR⁃497⁃5p and FBXW7 mRNA expressions were determined using qRT⁃PCR. The FBXW7 protein level was also detected using western blotting. The cell viability, migration and apoptosis were evaluated using CCK⁃8 assay, wound healing assay and flow cytometry analysis, respectively. Additionally, the binding relationships among lncRNA VIM⁃AS1, miR⁃497⁃5p and FBXW7 were verified by dual luciferase reporter assaies. Results: LncRNA VIM⁃AS1 and FBXW7 expressions were remarkably reduced in HG⁃treated ARPE⁃19 cells, while miR⁃497⁃5p was upregulated. LncRNA VIM ⁃ AS1 could upregulate the expression of FBXW7 by competitively binding to miR ⁃ 497 ⁃ 5p. LncRNA VIM ⁃ AS1 overexpression promoted cell proliferation and migration, and inhibited cell apoptosis in HG⁃induced ARPE⁃19 cells, while miR⁃497⁃ 5p overexpression abolished the effects of lncRNA VIM⁃AS1 overexpression on HG⁃induced ARPE⁃19 cells. Furthermore, FBXW7 knockdown abrogated the effects of miR⁃497⁃5p inhibition on cell phenotypes of HG⁃treated ARPE⁃19 cells. Conclusion: LncRNA VIM⁃AS1 could promote the proliferation and migration, while inhibited cell apoptosis of HG⁃treated ARPE⁃19 cells by regulation of miR⁃497⁃5p/FBXW7 axis, suggesting that lncRNA VIM⁃AS1 might have great potential as therapeutic target for diabetic retinopathy. [ABSTRACT FROM AUTHOR]

Objective To explore the effect of zerumbone on proliferation, migration and invasion of non-small cell lung cancer cell line and underline mechanism. Methods Human non-small cell lung cancer cells line A549 was divided into control group, zingerone-low dose group, zingerone-medium dose group, zingerone-high dose group, pcDNA group, pcDNA-FBXW7 group, and zingerone-high dose group +si-NC group, zingerone-high dose +si-FBXW7 group. Cell proliferation inhibition was detected by methyl thiazolyl tetrazolium (MTT) method, cell migration and invasion were detected by Transwell assays, and the expression of F-box with 7 tandem WD40 (FBXW7) mRNA and protein were detected by Western blot and real-time(RT-qPCR). Results Compared with control group, the proliferation, migration and invasion of A549 cells in the zingerone-low, medium and high dose groups were significantly reduced (P＜0.05), the expression of FBXW7 mRNA and protein were significantly increased (P＜0.05). Compared with the pcDNA group, the proliferation, migration and invasion of A549 cells in the pcDNA-FBXW7 group were significantly reduced (P＜0.05). Compared with the Zingerone-high-dose+si-NC group, the cell proliferation, migration and invasion ability of A549 in the Zingerone-high-dose+si-FBXW7 group was significantly increased (P＜0.05). Conclusions Zerumbone reduces proliferation, migration and invasion of non-small cell lung cancer cells, and the underlying mechanism is related to the up-regulation of FBXW7 expression.

Objective To investigate the effect of FBXW7 on the expression and localization of HSF1 in colorectal cancer cells. Methods The expression levels of HSF1 and pHSF1Ser326 protein in FBXW7 deletion (KO) and wild-type (WT) FBXW7-expressing counterpart colorectal cancer cells were detected by Western blot. The nucleoprotein expression and localization of pHSF1Ser326 in heat-shocked or recovery stage cells were observed by Western blot and immunofluorescence method. Results The HSF1 expression level in DLD1 cells transfected with FBXW7α was decreased significantly (P < 0.01). The expression levels of HSF1 and pHSF1Ser326 protein in FBXW7 KO cells were higher than those in WT cells (all P < 0.05). HSF1 and pHSF1Ser326 in FBXW7 KO cells were mainly expressed in nucleus and weakly expressed in cytoplasm. After warm stimulation, the expression of HSF1 and pHSF1Ser326 in WT cells recovered to the unstimulated level, while the expression of HSF1 and pHSF1Ser326 in FBXW7 KO cells were higher in the nucleus (all P < 0.01). Conclusion Loss of FBXW7 could affect the nuclear HSF1 recovery after warm stimulation. It may be associated with FBXW7 deletion inhibiting the degradation of nuclear HSF1.

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Objective To investigate the effects of microRNAs (miRNAs) on the proliferation and apoptosis of colorectal cancer HCT116 cells and its mechanism. Methods The FBXW7 mRNA and protein levels were detected by realtime quantitative PCR and Western blot assay in 20 clinical specimens. After predicted by TargetScan tool, verified by quantitative PCR and luciferase activity assay, the miRNAs combined with FBXW7 were identified. The FBXW7 mRNA and protein levels were detected after transient transfection of miR-223 and its control miCtr and inhibitor (inhibitor) into HCT116 cells. CCK-8 and flow cytometry were used respectively to detect the cell viability and apoptosis rate. In addition, the level of miR-223 and the number of apoptosis were detected after down-regulating the expression of Notch3 by siRNA. Results The FBXW7 mRNA and protein levels were lower in colorectal tumor tissues than those in adjacent tissues (P＜ 0.05). It was predicted and confirmed that miR-223 and miR-25 could specifically bind to FBXW7 gene. After transfection of miR-223 in HCT116 cells, the FBXW7 mRNA and protein levels were down-regulated (P＜0.01), the cell viability was increased (P＜0.05) and the number of apoptotic cells was decreased (P＜0.01). After down-regulation of Notch3 pathway, miR-223 mRNA levels decreased (P＜0.001), FBXW7 mRNA levels increased (P＜0.01) and apoptotic cells decreased (P＜ 0.05). Conclusion Notch pathway upregulates the miR-223 level, which inhibits the expression of FBXW7 and eventually promotes the proliferation and inhibits its apoptosis of colon cancer cell line HCT116. [ABSTRACT FROM AUTHOR]

Objective: To investigate the expression of microRNA-182 (miR-182) in the non-small cell lung cancer (NSCLC) tissues and its effect and mechanism on the proliferation of NSCLC cells. Methods: The expression of miR-182 in the malignant NSCLC tissue and adjacent normal tissue was estimated by quantitative reverse-transcription polymerase chain reaction (qRT-PCR). The Levels of FBXW7 and corresponding substrates were detected by western blot. Cell growth was detected by counting kit (CCK)-8 reagent and colony formation experiment. Cell cycle progression and cell apoptosis were determined through flow cytometry. The effect of miR-182 on the FBXW7 was determined using the Dual-Luciferase Reporter Assay System. Results: Compared with the adjacent normal tissues,the expression of miR-182 was significantly upregulated in the NSCLC tissues(P<0.05). Compared with the control group, the capacity of cell growth, colony formation capacity and cell cycle progression were dramatically increased, the apoptosis was decreased in the transfection group(P<0.05). In addition, the expression of FBXW7 protein was downregulated in the NSCLC tissus(P<0.05). The activity of luciferase in wild-type FBXW7 plasmid was significantly decreased by miRNA-182 mimics. However, the activity of luciferase hadn’t been affected by miRNA-182 mimics after mutation of binding site. Conclusions: The expression of miR-182 was up-regulated in the NSCLC tissues, which can promote the proliferation of NSCLC cells and participates in the occurrence and development of tumors by down-regulating the expression of FBXW7. [ABSTRACT FROM AUTHOR]