RNA干擾靶向沉默誘騙受體3基因增加人胰腺癌細胞的放射敏感性
發(fā)布時間:2018-02-08 23:28
本文關(guān)鍵詞: 胰腺癌 AsPC-細胞 誘騙受體基因 RNA干擾 放射敏感性 出處:《中國腫瘤生物治療雜志》2017年08期 論文類型:期刊論文
【摘要】:目的:探討RNA干擾沉默誘騙受體3(decoy receptor 3,DcR3)基因?qū)σ认侔┘毎派涿舾行缘挠绊懠捌湎嚓P(guān)機制。方法:構(gòu)建帶有DcR3 siRNA序列的穩(wěn)定表達質(zhì)粒,通過脂質(zhì)體轉(zhuǎn)染至胰腺癌AsPC-1細胞株,設對照組、siRNA(-)陰性對照組和DcR3 siRNA組,應用Western blotting檢測AsPC-1細胞中DcR3表達的變化,平板克隆形成實驗檢測轉(zhuǎn)染DcR3 siRNA后AsPC-1細胞放射敏感性的變化,流式細胞術(shù)檢測細胞凋亡,RT-PCR和Western blotting檢測Caspase-8、Caspase-3和PARP-1表達的變化。結(jié)果:DcR3 siRNA組細胞中DcR3蛋白表達水平較對照或siRNA(-)組明顯降低(均P0.01);DcR3 siRNA組的克隆形成率明顯低于對照或siRNA(-)組,其存活分數(shù)(survival fraction,SF)降低、α/β比值升高(均P0.01);放射后DcR3 siRNA組腫瘤細胞凋亡率明顯高于對照或siRNA(-)組(均P0.01);轉(zhuǎn)染DcR3 siRNA后可以明顯上調(diào)Caspase-8、Caspase-3的表達和下調(diào)PARP-1的表達。結(jié)論:RNA干擾沉默DcR3基因通過激活凋亡因子Caspase-8和Caspase-3促進細胞凋亡,從而增加胰腺癌細胞對放射的敏感性。
[Abstract]:Aim: to investigate the effect of RNA interference silencing receptor 3 decoy receptor 3 (DcR3) gene on radiosensitivity of pancreatic cancer cell line. Methods: stable expression plasmid with DcR3 siRNA sequence was constructed and transfected into AsPC-1 cell line of pancreatic carcinoma by liposome. Western blotting was used to detect the expression of DcR3 in AsPC-1 cells and the radiosensitivity of AsPC-1 cells transfected with DcR3 siRNA was detected by plate clone formation assay. RT-PCR and Western blotting were used to detect the expression of Caspase-8, Caspase-3 and PARP-1. Results the expression of DcR3 protein in the cells of the control group and the siRNA-treated group was significantly lower than that of the control or siRNA-treated group (P 0.01) and the clone formation rate of the siRNA group was significantly lower than that of the control group or siRNA-group. The survival fraction and 偽 / 尾 ratio of DcR3 siRNA group were significantly higher than those of control group or siRNA-group (P 0.01). After transfection of DcR3 siRNA, the expression of Caspase-8 and Caspase-3 and down-regulated expression of PARP-1 were significantly up-regulated. Conclusion the expression of Caspase-8 caspase-3 and down-regulated expression of PARP-1 can be down-regulated by DcR3 siRNA transfection (P < 0.01), and the apoptotic rate of tumor cells in DcR3 siRNA group after irradiation is significantly higher than that in control group or siRNA-group (P < 0.01). Silencing DcR3 gene promotes apoptosis by activating apoptosis factors Caspase-8 and Caspase-3. Thus increasing the radiosensitivity of pancreatic cancer cells.
【作者單位】: 湘南學院附屬醫(yī)院腫瘤中心;密蘇里大學醫(yī)學院Ellis
【基金】:湖南省自然科學基金資助項目(No.14JJ3136) 郴州市科技局科研基金資助項目(No.CZ2013096)~~
【分類號】:R735.9
【參考文獻】
相關(guān)期刊論文 前3條
1 項金峰;施思;梁丁孔;虞先o,
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