激光共聚焦显微镜观察^(60)Co γ射线对CNI-H446 DNA/RNA比值及三维结构的影响

Effects of irradiation with ^(60)Co γ-rays on the ratio of DNA and RNA in CNI-H446 cells and its three-dimensional structure by laser scanning confocal microscopy

目的 探讨电离辐射对小细胞肺癌细胞株 (CNI H4 4 6 )DNA RNA代谢比值的影响及观察细胞三维结构的改变。方法 利用共聚焦显微镜双重荧光标记、全自动三维图像分析CNI H4 4 6细胞受到不同剂量6 0 Coγ射线照射后DNA和RNA含量的比值。结果 对照组细胞DNA RNA比值为 1 15± 0 0 7,当受到 1Gy以上剂量照射时 ,DNA含量明显降低 ,诱导RNA合成逐渐增加 ,DNA降低的程度大于RNA ,1,3Gy组与对照组相比差异具有非常显著性 (P <0 0 1)。三维图像显示RNA聚集 ,RNA荧光强度明显增强 ,而DNA均匀散在于细胞内。当剂量 >8Gy时 ,RNA合成受到显著抑制 ,两者比值明显增加 ,细胞体积增大 ,其比值与照射剂量存在一定的联系。结论 CNI H4 4 6细胞受到不同剂量6 0 Coγ射线照射后DNA

Objective To investigate the effect of γ-radiation on DNA and RNA metabolism in cells of small cell lung cancer (SCLC) strain (CNI-H446) and their three-dimensional structure. Methods Laser scanning confocal microscopy (LSCM) and acridine orange (AO) staining was used to investigate the DNA/RNA changes in the cells,and automatic three-dimensional image scan was used to fine the change of its structure. Results The DNA/RNA of the control group was 1 15±0 07.When the dose of γ-radiation was more than 1 Gy,DNA decreased obviously and the extent of the decrease of DNA was larger than that of RNA.The values of DNA/RNA of the groups that were irradiated with 1 Gy and 3 Gy were significantly different from those of the control group ( P <0 01).The three-dimensional image manifested that after large dose of irradiation,the RNA in the cells gathered obviously while DNA were disrupted.When the dose was 8 Gy,the value of DNA/RNA was not different from that of the control group.When the dose was 8 Gy or higher,RNA was suppressed as seriously as DNA,and the value of DNA/RNA increased.There is certain relationship between the value of DNA/RNA and the dose of radiation. Conclusion After irradiated with different dose of γ-radiation,the ratio of DNA and RNA and the size of cells would change correspondingly.