^(125)I粒子治疗肝细胞癌的调控网络

Regulatory network of ^(125)I seeds for treatment of hepatocellular carcinoma

目的观察^(125)I粒子治疗肝细胞癌(HCC)的调控网络。方法以^(125)I粒子辐照HCC细胞(PLC/PRF/5细胞及HepG2细胞),通过平板克隆和免疫荧光染色实验检测辐照对HCC细胞的影响;以基因芯片筛选显著差异基因,采用Ingenuity Pathway Analysis(IPA)分析细胞信号通路及显著差异基因作用网络,并行qPCR及蛋白质印迹法检测,观察^(125)I粒子的调控网络。结果相比空白对照组,^(125)I粒子组PLC/PRF/5及HepG2细胞的平板克隆能力均显著降低,PLC/PRF/5细胞内γH2AX荧光明显增加。^(125)I粒子HCC细胞Acute Phase Response Signaling被抑制,106个基因上调,276个基因下调。qPCR及蛋白质印迹显示,辐照后HCC细胞乙酰基转移酶2(ESCO2)和双特异性磷酸酶1(DUSP1)表达下调,人DNA损伤诱导转录因子3(DDIT3)表达上调。结论^(125)I粒子治疗HCC的调控网络包括抑制Acute Phase Response Signaling、下调ESCO2表达使之无法及时修复细胞增殖时发生的DNA损伤,上调DDIT3表达、阻滞HCC细胞生长及下调DUSP1表达、诱导HCC细胞凋亡。

Objective To observe the regulatory network of ^(125)I seeds for treatment of hepatocellular carcinoma(HCC).Methods HCC cells(PLC/PRF/5 cells and HepG2 cells)were irradiated with ^(125)I seeds,and the impact of ^(125)I seeds on HCC cells was tested with colony forming assay and immunofluorescence staining.The significantly different genes were screened using gene chip,and signaling pathway and action network of significantly differential genes were analyzed with ingenuity pathway analysis(IPA).Then qPCR and Western blotting were performed,and the regulatory network of ^(125)I seeds was observed.Results Compared with blank control group,the plate cloning ability of PLC/PRF/5 and HepG2 cells of ^(125)I seeds group significantly reduced,whileγH2AX expression in PLC/PRF/5 cells of ^(125)I seeds group significantly increased.After ^(125)I seeds irradiation,Acute Phase Response Signaling was inhibited,and there were 106 up-regulated genes and 276 down-regulated genes.qPCR and Western blotting showed that the expression of establishment of sister chromatid cohesion N-acetytransferase 2(ESCO2)and dual specificity phosphatase 1(DUSP1)was down-regulated,while of DNA damage inducible transcript 3(DDIT3)was up-regulated after ^(125)I seeds irradiation.Conclusion The regulatory network of ^(125)I seeds for treatment of HCC included inhibited Acute Phase Response Signaling and down-regulated expression of ESCO2,resulting DNA damage during proliferation of HCC cells not be repaired in time,also up-regulated expression of DDIT3 that prevent the growth of HCC cells and down-regulated expression of DUSP1 that induce HCC cells apoptosis.