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目的:探讨Egr-1基因沉默对人肺腺癌A549细胞放射敏感性的影响。方法:选用A549细胞株作为研究对象,将其分成A、B、C三组,即空白对照组(只加入RPMI-1640培养)、阴性对照组(加入LV3-NC-sh RNA)、实验组(加入EGR1-homo-2294-sh RNA),采用慢病毒介导的sh RNA干扰技术使实验组细胞Egr-1基因沉默表达。利用荧光显微镜、自动化荧光定量细胞成像分析系统分析sh RNA转染,利用FQ-PCR分析Egr-1表达,再利用细胞克隆形成实验检测细胞放射敏感性参数的差异。结果:慢病毒介导的sh RNA成功转染阴性对照组、实验组细胞;空白对照组与阴性对照组Egr-1表达无差异(P>0.05),实验组与空白对照组、阴性对照组比较Egr-1均受到明显抑制(P<0.05);克隆形成实验中细胞放射敏感性参数D0、SF2实验组细胞与空白对照组、阴性对照组比较均存在明显差异(P<0.05)。结论:Egr-1基因沉默使A549细胞放射敏感性降低,Egr-1表达可能与肿瘤细胞对放射的敏感性有关。
Objective: To investigate the effect of Egr-1 gene silencing on the radiosensitivity of human lung adenocarcinoma A549 cells. Methods: The A549 cell line was selected as research object and divided into three groups: blank control group (only adding RPMI-1640 culture), negative control group (adding LV3-NC-sh RNA), experimental group EGR1-homo-2294-sh RNA was added), and the lentivirus-mediated sh RNA interference technique was used to silence the Egr-1 gene in the experimental group. The expression of Egr-1 was analyzed by FQ-PCR and the difference of radiosensitivity parameters was detected by cell clone formation assay. The fluorescent RNA was used to analyze sh RNA transfection by fluorescence quantitative cell imaging analysis system. Results: The lentivirus-mediated sh RNA was successfully transfected into the negative control group and the experimental group. The expression of Egr-1 in the blank control group and the negative control group was not significantly different (P> 0.05). Compared with the blank control group and the negative control group Egr-1 was significantly inhibited (P <0.05). There was significant difference in the cell radiosensitivity parameters D0 and SF2 between the control group and the blank control group (P <0.05). Conclusion: Egr-1 gene silencing reduces A549 cell radiosensitivity and Egr-1 expression may be related to the sensitivity of tumor cells to radiation.