本文關(guān)鍵詞： 小干擾RNA 真核細(xì)胞翻譯起始因子E 喉腫瘤 Hep-細(xì)胞 細(xì)胞凋亡　出處：《吉林大學(xué)學(xué)報(醫(yī)學(xué)版)》2014年01期 　論文類型：期刊論文

【摘要】：目的:觀察真核細(xì)胞翻譯起始因子4E(eIF4E)基因的靶向小干擾RNA(siRNA)對喉癌Hep-2細(xì)胞中eIF4E基因表達(dá)的影響,探討其誘導(dǎo)喉癌細(xì)胞凋亡的作用機制。方法:采用脂質(zhì)體LipofectamineTM2000將siRNA-eIF4E轉(zhuǎn)染入人喉癌Hep-2細(xì)胞(siRNA-eIF4E組),同時設(shè)空白對照組和空質(zhì)粒組。MTT法檢測siRNA對Hep-2細(xì)胞增殖的抑制作用,羅丹明染色檢測轉(zhuǎn)染后細(xì)胞內(nèi)線粒體膜電位的變化,TUNEL染色法檢測細(xì)胞凋亡,RT-PCR和Western blotting法檢測凋亡相關(guān)基因轉(zhuǎn)錄和蛋白表達(dá)水平的變化。結(jié)果:與空白對照組及空質(zhì)粒組比較,siRNA-eIF4E組Hep-2細(xì)胞中eIF4E基因轉(zhuǎn)錄和表達(dá)水平明顯下調(diào)(P0.01),Hep-2細(xì)胞生存率下降(P0.05),細(xì)胞線粒體膜電位降低(P0.05),細(xì)胞凋亡率增加(P0.05),凋亡相關(guān)基因Bim、Bid及Caspase-3表達(dá)上調(diào)(P0.05)。結(jié)論:siRNA-eIF4E在體外可抑制人喉癌Hep-2細(xì)胞增殖,其機制可能是通過激活線粒體凋亡途徑誘導(dǎo)Hep-2細(xì)胞凋亡。
[Abstract]:Aim: to observe the effect of targeting small interfering RNAs (siRNAs) of eukaryotic cell translation initiation factor 4EIF4E on the expression of eIF4E gene in Hep-2 cells of laryngeal carcinoma. Methods: siRNA-eIF4E was transfected into human laryngeal carcinoma Hep-2 cells by lipofectin LipofectamineTM2000 into siRNA-eIF4E group. The inhibitory effect of siRNA on the proliferation of Hep-2 cells was detected by blank control group and empty plasmid group. The changes of mitochondrial membrane potential after transfection were detected by Rhodamine staining and the expression of apoptosis-related genes by RT-PCR and Western blotting were detected by Tunel staining. Results: compared with the blank control group and empty substance, the changes of apoptosis-related gene transcription and protein expression were detected by Tunel staining. The transcription and expression of eIF4E gene in Hep-2 cells were significantly down-regulated by siRNA-eIF4E group. The survival rate of Hep-2 cells was decreased, the mitochondrial membrane potential was decreased, the apoptosis rate was increased, and the expression of the apoptosis-related gene BimsiRNA-eIF4E was increased. Conclusion\% siRNA-eIF4E can increase the expression of BimsiRNA-eIF4E in Hep 2 cells. It can inhibit the proliferation of human laryngeal carcinoma Hep-2 cells in vitro. The mechanism may be the activation of mitochondrial apoptosis pathway to induce apoptosis of Hep-2 cells.
【作者單位】： 吉林大學(xué)第二醫(yī)院耳鼻咽喉-頭頸外科;吉林大學(xué)基礎(chǔ)醫(yī)學(xué)院病理生理學(xué)系;長春中醫(yī)藥大學(xué)研發(fā)中心;
【基金】：國家自然科學(xué)基金資助課題(81302206) 吉林省科技廳自然科學(xué)基金資助課題(201105106)
【分類號】：R739.65

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