摘要
目的研究白杨素对肿瘤坏死因子-α(TNF-α)诱导慢性暴露转化生长因子-β(TGF-β)卵巢癌OVCAR-3细胞系球形成的影响,并探讨其作用机制是否涉及下调NF-κB(p65)蛋白表达。方法 TNF-α(10μg·L^(-1))处理TGF-β(5μg·L^(-1))预暴露12 d的卵巢癌OVCAR-3细胞24 h,球形成率测定法检测不同浓度(5.0、10.0、20.0μmol·L^(-1))白杨素对球形成率的影响;Western blot检测NF-κB(p65)蛋白表达;NF-κB(p65)siRNA转染探讨作用机制。结果 TNF-α(10μg·L^(-1))联合慢性暴露TGF-β(5μg·L^(-1))处理增高OVCAR-3细胞系球形成率。白杨素能有效地拮抗致炎因子诱导卵巢癌细胞自我更新作用,呈剂量依赖性(P<0.05),并伴随着NF-κB(p65)蛋白表达下调。与对照siRNA相比,NF-κB(p65)siRNA转染OVCAR-3细胞NF-κB(p65)蛋白表达下调,并明显增强白杨素抑制球形成作用。结论下调NF-κB(p65)蛋白表达参与白杨素抑制TNF-α(10μg·L^(-1))联合慢性暴露TGF-β(5μg·L^(-1))处理诱导卵巢癌OVCAR-3细胞系球形成作用。
Aim To investigate the effect of chrysin（ ChR） on sphere formation of ovarian cancer OVCAR -3 cell line induced by treatment with tumor necrosisfactor-a（TNF-a） following chronic exposure to transforminggrowth factor-β（ TGF-β ） ,and its mechanismunderlying whether this action is involved in down-regulationof NF-κB （p65） protein expression. MetliodsAfter pre-exposure to TGF-β （5μg·L^-1 ） for 12 d ,OVCAR-3 cells were treated with TNF-α （ 10μg·L^-1） for 24 h , tumor sphere-forming assay was used todetect the rate of tumor sphere formation of OVCAR-3cells treated with or without various concentrations ofChR（5.0 , 10.0 and 20.0μmol·L^-1）. The proteinexpression of NF-κB （p65） was analyzed using Westernblot. Transfection with NF-κB （ p65 ） siRNA was usedto explore the mechanism by which C h R inhibited selfrenewof OVCAR-3 cells. Results After co-treatmentof TNF-α（10μg·L^-1） and chronic exposure to TGF-β （5μg·L^-1）, the rate of sphere formation of ovariancancer OVCAR-3 cells was increased. C h R effectivelyreduced sphere-forming capacity of ovarian cancer cellsinduced by inflammatory factor in a dose dependentmanner（P 〈0. 05） , and the protein expression level of NF-κB（p65） was down-regulated. Compared with thecells transfected with control siRNA,the expression of NF-κB （ p65 ） protein was down-regulated in cellstransfected with NF-κB（p65） siRNA,and the NF-κB（ p65 ） siRNA significantly enhanced the inhibitoryeffect of C h R on the sphere -forming capacity of OVCAR-3 cells. Conclusions The inhibitory effects of ChR on sphere-forming capacity of ovarian cancerOVCAR-3 cells induced by co-treatmemt of TNF-cawith chronic exposure to TGF-β pare involved in downregulationof NF-κB（p65） protein expression.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2016年第8期1133-1137,共5页
Chinese Pharmacological Bulletin
基金
湖南师范大学青年科学基金资助项目(No医140666)
国家自然科学基金面上项目(No 81172375)
