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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 investigate the effect of interferon inducible protein 16 (IFI16) on the proliferation of basic fibroblast growth factor (bFGF) -induced vascular endothelial cells (VECs) and its possible mechanism. Methods Human umbilical vein endothelial cells were cultured and divided into negative control group, IFN-α induction group and IFI16 siRNA interference group, IFN-α was stimulated with bFGF (final concentration of 100 μg / L), siRNA of siRNA was transfected into siRNA of IFI16 for 24 h, 24 h, 48 h and 72 h after intervention respectively. Cell viability was measured by MTT assay The cell cycle was analyzed by flow cytometry. The mRNA and protein expression of IFI16 and p21 were detected by semi-quantitative RT-PCR and Western blot, respectively. Meanwhile, the expression of Ras protein was also observed. Results IFN-α induced up-regulation of VEC IFI16 mRNA and protein expression, decreased VEC cell viability and cell cycle G1 / S conversion [8.16 ± 1.10% (13.84 ± 0.92)%], with p21 mRNA and protein expression Upregulation and down-regulation of Ras protein. Transfection of IFI16 siRNA inhibited the expression of IFI16 and p21, promoted the expression of Ras protein, increased the VEC cell viability and promoted the G1 / S transition of cell cycle [17.03 ± 0.52% (13.84 ± 0.92)%]. Conclusion IFI16 can inhibit the proliferation of VEC induced by bFGF, which may be related to the activation of p21 and the inhibition of Ras expression.