抑制干擾素誘導(dǎo)蛋白16表達(dá)對(duì)人腦血管外膜成纖維細(xì)胞增殖、凋亡及遷移的影響
發(fā)布時(shí)間：2018-10-22 06:51
【摘要】：目的探討抑制干擾素誘導(dǎo)蛋白16(IFI16)表達(dá)后對(duì)人腦血管外膜成纖維細(xì)胞(HBVAF)增殖、凋亡與遷移的影響及其可能機(jī)制。方法培養(yǎng)HBVAF分3組,未經(jīng)處理的HBVAF作為空白對(duì)照組,轉(zhuǎn)染非特異性小干擾RNA(siRNA)的HBVAF作為陰性對(duì)照組,轉(zhuǎn)染IFI16siRNA的HBVAF作為IFI16siRNA組。應(yīng)用蛋白免疫印跡和實(shí)時(shí)熒光定量聚合酶鏈反應(yīng)(PCR)測(cè)定細(xì)胞中IFI16、p53、p21蛋白和mRNA表達(dá)水平。用四甲基偶氮唑鹽(MTT)比色法檢測(cè)細(xì)胞活力,流式細(xì)胞術(shù)測(cè)定細(xì)胞凋亡,Transwell法測(cè)定細(xì)胞遷移情況。結(jié)果與陰性對(duì)照組比較,轉(zhuǎn)染IFI16siRNA后,HBVAF中IFI16、p53及p21蛋白和mRNA表達(dá)水平下調(diào),IFI16siRNA組MTT吸光度值增高(0.70±0.01比0.65±0.01,P0.05),IFI16siRNA組、陰性對(duì)照組與空白對(duì)照組之間在細(xì)胞遷移數(shù)目與凋亡比例上差異無統(tǒng)計(jì)學(xué)意義。結(jié)論抑制IFI16表達(dá)可促進(jìn)HBVAF增殖,其機(jī)制可能部分與抑制p53及p21表達(dá)有關(guān)。
[Abstract]:Objective to investigate the effect of inhibiting the expression of interferon inducible protein 16 (IFI16) on the proliferation, apoptosis and migration of human cerebrovascular adventitial fibroblasts (HBVAF) and its possible mechanism. Methods HBVAF were divided into three groups: untreated HBVAF as blank control group, HBVAF transfected with non-specific small interfering RNA (siRNA) as negative control group and IFI16siRNA HBVAF as IFI16siRNA group. Western blot and real-time fluorescence quantitative polymerase chain reaction (PCR) were used to detect the expression of IFI16,p53,p21 protein and mRNA. Cell viability was detected by (MTT) assay, apoptosis was detected by flow cytometry, and cell migration was measured by Transwell assay. Results compared with the negative control group, the expression of IFI16,p53 and p21 protein and mRNA in HBVAF were down-regulated after transfection of IFI16siRNA, and the absorbance of MTT in IFI16siRNA group was increased (0.70 鹵0.01 vs 0.65 鹵0.01 P 0.05). There was no significant difference in the number of cell migration and the proportion of apoptosis between the negative control group and the blank control group. Conclusion inhibiting the expression of IFI16 can promote the proliferation of HBVAF, and the mechanism may be related to the inhibition of p53 and p21 expression.
【作者單位】： 貴州省人民醫(yī)院心內(nèi)科;
【基金】：國家自然科學(xué)基金項(xiàng)目(81260030)
【分類號(hào)】：R54
本文編號(hào)：2286411
[Abstract]:Objective to investigate the effect of inhibiting the expression of interferon inducible protein 16 (IFI16) on the proliferation, apoptosis and migration of human cerebrovascular adventitial fibroblasts (HBVAF) and its possible mechanism. Methods HBVAF were divided into three groups: untreated HBVAF as blank control group, HBVAF transfected with non-specific small interfering RNA (siRNA) as negative control group and IFI16siRNA HBVAF as IFI16siRNA group. Western blot and real-time fluorescence quantitative polymerase chain reaction (PCR) were used to detect the expression of IFI16,p53,p21 protein and mRNA. Cell viability was detected by (MTT) assay, apoptosis was detected by flow cytometry, and cell migration was measured by Transwell assay. Results compared with the negative control group, the expression of IFI16,p53 and p21 protein and mRNA in HBVAF were down-regulated after transfection of IFI16siRNA, and the absorbance of MTT in IFI16siRNA group was increased (0.70 鹵0.01 vs 0.65 鹵0.01 P 0.05). There was no significant difference in the number of cell migration and the proportion of apoptosis between the negative control group and the blank control group. Conclusion inhibiting the expression of IFI16 can promote the proliferation of HBVAF, and the mechanism may be related to the inhibition of p53 and p21 expression.
【作者單位】： 貴州省人民醫(yī)院心內(nèi)科;
【基金】：國家自然科學(xué)基金項(xiàng)目(81260030)
【分類號(hào)】：R54
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