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目的:探讨H IV-1 Tat蛋白对细胞周期相关基因表达以及电离辐射诱发细胞周期阻滞的影响。方法:使用包含102个与DNA损伤修复和细胞周期相关的基因微阵列检测人横纹肌肉瘤细胞(TE671)及已转染tat基因的TE671细胞(TT2)基因表达谱的改变;使用流式细胞仪检测细胞周期变化;W estern印迹检测蛋白表达变化。结果:在基因芯片的检测中发现,与DNA损伤修复及细胞周期调控相关的6个基因Cdc25C,K IF2C,Cdc20,DNA-PKcs,CTS1,W ee1在转染tat基因的细胞中表达下调;细胞周期检测发现TE671细胞和TT2细胞经4 Gyγ射线照射后表现出不同程度的G2/M期阻滞,但表达Tat的TT2细胞G2/M阻滞出现较TE671细胞晚,而在照射后48 h时TE671细胞G2/M期阻滞已恢复,但TT2细胞阻滞仍很显著。另外,TT2细胞S期阻滞时间延长。研究进一步发现细胞周期蛋白(cyc lin)B1在TT2细胞中表达增强。结论:H IV-1Tat蛋白导致G2/M检验点功能紊乱,将影响细胞的辐射敏感性,本研究为了解AIDS合并肿瘤患者对放射治疗敏感性提供了重要实验数据。
Objective: To investigate the effects of H IV-1 Tat on cell cycle related gene expression and ionizing radiation-induced cell cycle arrest. Methods: The gene expression profiles of TE671 cells and TE671 cells (TT2) of human rhabdomyosarcoma cells (TE671) and the transfected tat gene were detected by using 102 microarray including DNA microbiology and cell cycle-related gene microarrays. Flow cytometry Cell cycle changes; Western blotting detected protein expression changes. Results: Six genes Cdc25C, KIF2C, Cdc20, DNA-PKcs, CTS1 and W ee1 related to DNA damage repair and cell cycle regulation were down-regulated in the gene transfected cells. Cytometry showed that G2 / M arrest in TE671 cells and TT2 cells after irradiation with 4 Gy γ-rays showed different degrees of G2 / M arrest, whereas G2 / M arrest in TT2 cells expressing Tat appeared later than that of TE671 cells. However, at 48 h after irradiation, The G2 / M arrest of TE671 cells has been restored, but the arrest of TT2 cells is still significant. In addition, TT2 cells S-phase prolongation. The study further found that cyclin B1 expression in TT2 cells increased. Conclusion: H IV-1Tat protein causes G2 / M checkpoint dysfunction and affects cell radiation sensitivity. This study provides important experimental data to understand the sensitivity of patients with AIDS-associated tumors to radiotherapy.