论文部分内容阅读
目的探讨照射剂量和照射时间对电离辐射诱导L02人正常肝细胞(简称“L02细胞”)的线粒体编码基因细胞色素c氧化酶(COX)Ⅱ基因表达、三磷酸腺苷(ATP)水平和细胞活力变化的影响。方法 采用2×7析因设计方法。0、1、3、5、8、10、15 Gy剂量60Coγ射线分别照射L02细胞24、48 h后,采用逆转录聚合酶链反应(PCR)和实时荧光定量PCR法检测COXⅡmRNA表达水平,蛋白质印迹法检测COXⅡ蛋白表达水平,ATP发光检测试剂盒和CCK-8试剂盒检测细胞内ATP水平和细胞活力。结果 照射时间与照射剂量对L02细胞的COXⅡmRNA及蛋白表达水平上调、ATP水平升高和细胞活力下降的影响存在交互作用(F值分别为92.43、267.40、6.99、116.11,P<0.05或P<0.001)。在一定照射剂量范围内,照射后24 h COXⅡ蛋白表达水平、照射后24、48 h ATP水平和细胞活力分别与照射剂量存在剂量-效应关系(P<0.05或P<0.01)。结论 照射剂量和照射时间的交互作用影响60Coγ照射诱导的L02细胞COXⅡ基因表达和细胞内ATP水平与细胞活力。
Objective To investigate the effect of irradiation dose and irradiation time on the expression of cytochrome c oxidase (COX) Ⅱ gene, the level of ATP and the cell viability in mitochondria of L02 human normal hepatocytes (“L02 cells”) under ionizing radiation Impact. Methods 2 × 7 factorial design method. L02 cells were irradiated with 60Coγ ray at dosages of 0, 1, 3, 5, 8, 10 and 15 Gy for 24 and 48 h respectively. The expression of COX Ⅱ mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR) and real- COXⅡprotein expression was detected by enzyme-linked immunosorbent assay (ELISA), and the ATP level and cell viability were detected by ATP Luminescence assay and CCK-8 assay. Results There was interaction between COX Ⅱ mRNA and protein expression level, ATP level and cell viability in L02 cells (F = 92.43, 267.40, 6.99, 116.11 respectively, P <0.05 or P <0.001 ). The COXⅡprotein expression at 24 h after irradiation and the dose-response relationship (P <0.05 or P <0.01) between the levels of ATP and cell viability at 24 h and 48 h after irradiation were dose-dependent and dose-dependent, respectively. Conclusion The interaction between irradiation dose and irradiation time affects COXⅡ gene expression and intracellular ATP level and cell viability induced by 60Coγ irradiation in L02 cells.