Akimoto T, Mitsuhashi N, Saito Y, Ebara T, Niibe H
Department of Radiology and Radiation Oncology, Gunma University School of Medicine, Maebashi, Japan.
Int J Radiat Oncol Biol Phys. 1998 Jul 15;41(5):1171-6. doi: 10.1016/s0360-3016(98)00176-x.
To investigate the effect of radiation on E-cadherin and alpha-catenin expression in a human lung cancer cell line, and also evaluate invasive capacity in the membrane invasion culture system using the Boyden Chamber.
The immunoblot and immunofluorescence analyses were performed using the human lung cancer cell line A549 to examine altered expression of E-cadherin and alpha-catenin after irradiation. We also compared invasive capacity of untreated cells with that of irradiated cells.
Immunoblot analysis revealed that the expression of E-cadherin increased after irradiation. In a time-course analysis, the expression was increased 6 h after irradiation with 10 Gy and reached its peak level at 24 h, being 2.3 times the control value, whereas expression at 1 and 3 h after irradiation was almost equivalent to that of the control. A slight increase in expression was observed after irradiation of 2 Gy and the expression reached peak levels after 5 Gy. After fractionated irradiation, the increase in expression of both E-cadherin and alpha-catenin was observed, and the alteration of alpha-catenin was more prominent than that after a single irradiation of the same total dose. In the immunofluorescence study for E-cadherin antibody analyzed by confocal laser scanning microscopy, increased intensity in irradiated cells produced as a nondisrupted and continuous line at cell-cell contact sites. In an invasive assay, the number of migrated cells in irradiated cells after a dose of 5 and 10 Gy was reduced significantly compared to untreated cells.
The results indicate that irradiation of A549 increased the expression of E-cadherin, possibly preserving their functional property.
研究辐射对人肺癌细胞系中E-钙黏蛋白和α-连环蛋白表达的影响，并使用博伊登小室在膜侵袭培养系统中评估侵袭能力。
使用人肺癌细胞系A549进行免疫印迹和免疫荧光分析，以检测辐射后E-钙黏蛋白和α-连环蛋白表达的变化。我们还比较了未处理细胞与辐射后细胞的侵袭能力。
免疫印迹分析显示，辐射后E-钙黏蛋白的表达增加。在时间进程分析中，用10 Gy辐射后6小时表达增加，并在24小时达到峰值水平，为对照值的2.3倍，而辐射后1小时和3小时的表达几乎与对照相当。2 Gy辐射后观察到表达略有增加，5 Gy辐射后表达达到峰值水平。分次照射后，观察到E-钙黏蛋白和α-连环蛋白的表达均增加，且α-连环蛋白的变化比相同总剂量单次照射后更明显。在通过共聚焦激光扫描显微镜分析的E-钙黏蛋白抗体免疫荧光研究中，辐射细胞中强度增加，在细胞间接触部位形成不间断的连续线。在侵袭试验中，与未处理细胞相比，5 Gy和10 Gy剂量辐射后细胞的迁移细胞数量显著减少。
结果表明，A549细胞经辐射后E-钙黏蛋白表达增加，可能保留了其功能特性。
