Objective To investigate the effect of inhibiting stearoyl-CoA desaturase 1(SCD1)activity on the biological behavior of cervical cancer cells and elucidate the molecular mechanisms related to fatty acid metabolism.Methods Cancer tissues were collected from 48 patients with cervical cancer,whereas normal cervical tissues were obtained from 48 patients with uterine fibroids.Immunohisto-chemical SP staining and real-time quantitative PCR were performed to assess differences in SCD1 expression.The MTS assay was used to determine cell viability,and EdU staining was conducted to evaluate cell proliferation.Cell migration and invasion abilities were analyzed using in vitro scratch assay and Transwell chamber assay.Immunofluorescence staining was performed to detect E-cadherin and vimentin expression.Real-time quantitative PCR was used to measure the mRNA expression levels of SREBP1,ACLY,ACC,and FAS.Nile red fluorescence staining was applied to observe intracellular lipid droplet content.Results The positive expression rate of SCD1 and the relative expression level of SCD1 mRNA were significantly higher in cervical cancer tissues than in normal cervical tissues(P＜0.05).Treatment of HeLa cells with varying concentrations of MF-438 led to significant reductions in cell viability,EdU-positive cell rate,scratch closure rate,and the number of migrating and invading cells.Additionally,the fluorescence intensity of E-cadherin increased,whereas that of vimentin decreased.The relative expression levels of SREBP1,A CL Y,ACC,and FAS mRNA were downregulated,and intracellular lipid droplet content decreased in a concentration-dependent manner(P＜0.05).Conclusion Inhibition of SCD1 activity significantly reduced lipid metabolism in HeLa cells and suppressed malignant biological behaviors,including proliferation,migration,invasion,and epithelial-mesenchymal transition.

Objective To investigate the effect of inhibiting stearoyl-CoA desaturase 1(SCD1)activity on the biological behavior of cervical cancer cells and elucidate the molecular mechanisms related to fatty acid metabolism.Methods Cancer tissues were collected from 48 patients with cervical cancer,whereas normal cervical tissues were obtained from 48 patients with uterine fibroids.Immunohisto-chemical SP staining and real-time quantitative PCR were performed to assess differences in SCD1 expression.The MTS assay was used to determine cell viability,and EdU staining was conducted to evaluate cell proliferation.Cell migration and invasion abilities were analyzed using in vitro scratch assay and Transwell chamber assay.Immunofluorescence staining was performed to detect E-cadherin and vimentin expression.Real-time quantitative PCR was used to measure the mRNA expression levels of SREBP1,ACLY,ACC,and FAS.Nile red fluorescence staining was applied to observe intracellular lipid droplet content.Results The positive expression rate of SCD1 and the relative expression level of SCD1 mRNA were significantly higher in cervical cancer tissues than in normal cervical tissues(P＜0.05).Treatment of HeLa cells with varying concentrations of MF-438 led to significant reductions in cell viability,EdU-positive cell rate,scratch closure rate,and the number of migrating and invading cells.Additionally,the fluorescence intensity of E-cadherin increased,whereas that of vimentin decreased.The relative expression levels of SREBP1,A CL Y,ACC,and FAS mRNA were downregulated,and intracellular lipid droplet content decreased in a concentration-dependent manner(P＜0.05).Conclusion Inhibition of SCD1 activity significantly reduced lipid metabolism in HeLa cells and suppressed malignant biological behaviors,including proliferation,migration,invasion,and epithelial-mesenchymal transition.

Objective To optimize the culture method for rheumatoid arthritis fibroblast-like synoviocytes in vitro,and observe the effect of Dishevelled (Dvl) 2 on vascular endothelial growth factor (VEGF) in rheumatoid arthritis fibroblast-like synoviocytes (RA-FLS).Methods Synovium from RA patients who underwent knee arthroplasties were cut into small piece,and RA-FLS were isolated and cultured in vitro using tissue block method.Dvl 2 lentivirus overexpressing plasmid was constructed and transfected into RAFLS.Q-polymerase chain reaction (PCR) and enzyme linked immunosorbent assay (ELISA) were used to detect the mRNA and protein expression levels of VEGF.Then we used 10 ng/ml tumor necrosis factor (TNF)-α recombinant protein to stimulate the transfected RA-FLS.24 h after stimulation,mRNA and protein expression of VEGF were detected again.Student's t test was used for two group analyses.Results RA-FLS was successfully isolated and cultured in vitro.The multiplicity of infection was 30 and was in conjunction with appropriate concentration of polybrene to promote transfection.Transfection efficiency could meet the test requirements.The mRNA of Dvl 2 increased for 79-fold than the control group.Compared with the control group,Dvl 2 could mildly inhibit RA-FLS secretion of VEGF.After TNF-α stimulation,Dvl 2 could significantly inhibit the VEGF's mRNA (2.15±0.10,2.92±0.47 fold,t=-3.924,P=0.003) and protein [(285±100) pg/ml,(155±61) pg/ml,t=-2.714,P=0.022] expression compared with the control group.Conclusion Dvl 2 can inhibit the effect of TNF-α induced secretion of VEGF in RA-FLS.The specific mechanism needs further study.