ROS影响有丝分裂时期细胞内线粒体的形态及分布

Morphology and location of mitochondria during mitosis impacted by ROS

目的利用稳定表达线粒体荧光信号的细胞系,检测活性氧(reactive oxygen species,ROS)对有丝分裂时期细胞的线粒体形态及分布的影响。方法构建表达线粒体荧光信号的病毒表达载体,经测序鉴定和Western印迹检测正确后,通过病毒包装及感染的方法构建稳定表达线粒体荧光信号的He La s3细胞系He La s3-COX4tp-EGFP;进一步用免疫荧光法观察线粒体在ROS影响下形态及分布的变化。结果成功构建表达线粒体荧光信号的病毒表达载体及稳定细胞系,通过激光共聚焦显微镜观察,发现H2O2处理能显著影响有丝分裂时期细胞线粒体的形态及分布。结论初步证明ROS可影响线粒体有丝分裂时期的形态及分布,为后续研究线粒体功能与细胞有丝分裂的调控奠定基础。

Objective To detect the impact of reactive oxygen species（ ROS） on mitochondrial morphology and distribution during mitosis. Methods A viral vector in which the fluorescence gene was specifically under the control of mitochondrial promoter was constructed and confirmed through DNA sequencing and Western blotting. After transfecting HeLa s3 cell with packaged virus,the HeLa s3-COX4tp-EGFP cell line stably expressing the mitochondrial fluorescence signal was obtained. With immunofluorescent staining,the impact of ROS on the morphology and distribution of mitochondria during mitosis was inspected. Result The cell line constantly expressing mitochondrial fluorescence signals was successfully constructed. Meanwhile,it was found that H2O2 treatment could significantly change the morphology and distribution of mitochondria during mitosis by confocal microscopy. Conclusion Our study demonstrates that ROS can affect the morphology and distribution of mitochondria during mitosis. This research help study the relationship between the mitochondrial function and the regulation of mitosis in the future.