RNA干扰靶向沉默诱骗受体3基因增加人胰腺癌细胞的放射敏感性

发布时间：2018-02-08 23:28

本文关键词： 胰腺癌 AsPC-细胞诱骗受体基因 RNA干扰放射敏感性　出处：《中国肿瘤生物治疗杂志》2017年08期 　论文类型：期刊论文

【摘要】：目的:探讨RNA干扰沉默诱骗受体3(decoy receptor 3,DcR3)基因对胰腺癌细胞放射敏感性的影响及其相关机制。方法:构建带有DcR3 siRNA序列的稳定表达质粒,通过脂质体转染至胰腺癌AsPC-1细胞株,设对照组、siRNA(-)阴性对照组和DcR3 siRNA组,应用Western blotting检测AsPC-1细胞中DcR3表达的变化,平板克隆形成实验检测转染DcR3 siRNA后AsPC-1细胞放射敏感性的变化,流式细胞术检测细胞凋亡,RT-PCR和Western blotting检测Caspase-8、Caspase-3和PARP-1表达的变化。结果:DcR3 siRNA组细胞中DcR3蛋白表达水平较对照或siRNA(-)组明显降低(均P0.01);DcR3 siRNA组的克隆形成率明显低于对照或siRNA(-)组,其存活分数(survival fraction,SF)降低、α/β比值升高(均P0.01);放射后DcR3 siRNA组肿瘤细胞凋亡率明显高于对照或siRNA(-)组(均P0.01);转染DcR3 siRNA后可以明显上调Caspase-8、Caspase-3的表达和下调PARP-1的表达。结论:RNA干扰沉默DcR3基因通过激活凋亡因子Caspase-8和Caspase-3促进细胞凋亡,从而增加胰腺癌细胞对放射的敏感性。
[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 伪 / 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.
【作者单位】：湘南学院附属医院肿瘤中心;密苏里大学医学院Ellis
【基金】：湖南省自然科学基金资助项目(No.14JJ3136) 郴州市科技局科研基金资助项目(No.CZ2013096)~~
【分类号】：R735.9

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