摘要
目的观察干扰素诱导蛋白16(IFI16)对碱性成纤维细胞生长因子(bFGF)诱导的血管内皮细胞(VEC)增殖的影响及可能机制。方法培养人脐静脉内皮细胞,分为阴性对照组(对照组)、α干扰素(IFN-α)诱导组(IFN组)和IFI16小干扰RNA(siRNA)干扰组(siRNA组),各组均用bFGF(终浓度为100μg/L)刺激24h后IFN组加IFN-α、siRNA组转染IFI16基因的siRNA,分别干预6h,干预后24、48和72h,用MTT法测定细胞活力反映细胞增殖,流式细胞仪分析细胞周期,用半定量RT-PCR法和Western blot分别检测IFI16和p21 mRNA和蛋白表达,同时观察Ras蛋白的表达。结果 IFN-α可诱导VEC IFI16 mRNA和蛋白表达上调,降低VEC细胞活力和细胞周期G1/S转换[S期细胞(8.16±1.10)%比(13.84±0.92)%],伴p21 mRNA和蛋白表达上调及Ras蛋白表达下调。转染IFI16siRNA可抑制IFI16和p21表达,促进Ras蛋白表达,提高VEC细胞活力和促进细胞周期G1/S转换[S期细胞(17.03±0.52)%比(13.84±0.92)%]。结论 IFI16表达可抑制bFGF诱导的VEC的增殖,该效应可能与激活p21及抑制Ras的表达有关。
Objective To observe the effect of interferon-inducible protein 16（IFI16） on the proliferation of vascular endothelial cells （VECI induced by basic fibroblast growth factor （bFGF）. Methods Human umbilical vein endo- thelial cells （HUVEC） were cultured and divided into three groups, control group, interferon alpha group （IFN group） and small interference RNA targeting IFI16 gene group（siRNA group）. Cultured VEC was stimulated by 100μg/L bFGF in every group foilowed by IFN-α or Ifil6 siRNA intervention for 6h in IFN-α or siRNA group respectively. 24, 48 and 79. h after intervention, cells vitality was detected by MTT assay, cell cycle was analyzed by flow cytometry, the mRNA and protein expression of IFI16 and p21 were detected by semiquantitative RT-PCR and Western blot, the expression of Ras protein was detected by Western blot. Results In VEC, IFN-α significantly increased the expression of IFI16 at mRNA and protein levels, reduced the cell vitality and cell cycle transition at G1/S [-S （ 8. 16±1.10 ） % vs （ 13.84±0.92 ） % ]. Meanwhile, stimulated with IFN-α up-regulated the expression of p21, down-regulated the expression of Ras. Transfection of Ifi16 siRNA inhibited the expression of IFI16 and p21, increased the expression of Ras protein and the cell vitality, promoted cell cycle transition at G1/S [S（ 17.03±0.52） % vs （13.84±0.92） %]. Conclusion The increased expression of IFI16 inhibits the proliferation of VEC induced by bFGF, which may be related to the activating of p21 and suppressing of Ras expression.
出处
《中华高血压杂志》
CAS
CSCD
北大核心
2013年第1期29-33,共5页
Chinese Journal of Hypertension
基金
贵州省高层次人才科研条件特助经费项目(TZJF-2010-098)
