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Objective@#To explore the role and underlying molecular mechanisms of fibroblast growth factor 21 (FGF21) in the development of melanoma.@*Methods@#Quantitative RT-PCR was applied to investigate the mRNA levels of FGF21 in cells. FGF21 overexpression plasmid was used to transfect FGF21 in melanoma cells, and shRNA technology was used to knock down FGF21 expression in melanoma cells. CCK8 assay and BrdU staining were used to detect the abilities of cell growth and proliferation. RT-qPCR was used to evaluate the expression levels of fat acids-oxidation-associated genes PGC1a, CD36, CPT1a, CPT1b and CPT2. Melanoma cells with or without FGF21 overexpression were subcutaneously injected into nude mice to observe the tumorigenic ability of the cells.@*Results@#The mRNA expression level of FGF21 was about 10 times higher than that of melanocytes (P<0.05). After the specific knockdown of FGF21 or overexpression of FGF21 in melanoma cells, cell proliferation levels were significantly reduced or increased (P<0.05). In vitro experiments showed that the overexpression of FGF21 significantly promoted the fatty acid oxidation in melanoma cells in terms of increased PGC1a, CD36, CPT1a, CPT1b and CPT2 gene expression. Further studies found that after the inhibition of intracellular fatty acid oxidation by small molecule Etomoxir, the cell proliferation ability of Etomoxir group was significantly lower than that of the control group by CCK8 assay and BrdU staining (P<0.05). Meanwhile, the tumor growth of the nude mice with FGF21-overexpressed melanoma cells was dramatically enhanced, and the tumor diameters were significantly increased.@*Conclusions@#FGF21 could promote the growth and proliferation of melanoma cells through regulating intracellular fatty acid oxidation, which accelerates the deterioration of melanoma.
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Objective TostudytheeffectofmicroRNA(miRNA) on the expression ofVEGF in ovarian cancer cells and its effect on tumor cell proliferation,in order to provide a new way for clinical diagnosis and treatment.Methods The two transfected cells were divided into HO-8910-H1-VEGF group and HO-8910-H1 group,and untransfected cells were referred to as the control group(HO-8910 group) by MTr(tetrazolium salt) method.The effect of miRNA on the proliferation of ovarian cancer cells was observed by periodic analysis.Results The expression of VEGF in HO-8910 cells was significantly decreased after transfection with specific small hairpin miRNA plasmid expression vector,ie,the expression of VEGF in HO-8910 H1-VEGF group was decreased compared with HO-8910 H1 group and HO-8910 group;Indirect immunofluorescence assay showed that the HO-8910-H1-VEGF group showed weak fluorescence intensity,but the HO-8910 H1 group and HO-8910 group had stronger fluorescence intensity.Cells in the HO-8910-H1-VEGF group that were blocked in the phase G1 (inter val phase 1) increased by 24％,and cells in phase S (DNA synthesis phase) showed a 21％ reduction.Conclusion MicroRNA can significantly inhibit the expression of VEGF in ovarian cancer HO-8910 cells,thereby blocking the proliferation of ovarian cancer cells and providing a theoretical basis for clinical gene therapy.
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Objective To explore the molecular mechanism by which YY1 associated factor 2(YAF2) up-regulates cyclin D1 expression in tumor cells as well as the effect of YAF2-cyclin D1 regulatory loop on tumor cell prolifera-tion. Methods Overexpression and knockdown experiments combined with Western blot and real-time quantitative PCR were used to detect the expression of YAF2 and cyclin D1;Dual luciferase reporter assay was performed to in-vestigate the effect of YAF2 on cyclin D1 promoter activity;Flow cytometry analysis was carried out to elucidate the effect of YAF2 on cell cycle progression through targeting cyclin D1;Colony formation assay was employed to deter-mine cell proliferation under different YAF2 and cyclin D1 expression level. Results YAF2 upregulated the ex-pression of cyclin D1 at both the mRNA(P<0.05) and protein level;YAF2 activated the promoter activity of cyc-lin D1 (P<0.05);YAF2 silencing increased significantly the proportion of cells in G0/G1phase(P<0.0001) and reduced the proportion of cells in S phase by regulating cyclin D1 (P<0.002); YAF2 facilitated the cell colony formation via targeting cyclin D1(P<0.05). Conclusions In tumor cells,YAF2 promotes the expression of cyclin D1,and enhances cell cycle progression and cell proliferation.