工频磁场对中国仓鼠肺成纤维细胞F-actin细胞骨架及其相关信号分子的影响

发布时间：2019-02-23 22:27

【摘要】： 随着电力事业的发展,极低频磁场(extremely low frequency magnetic fields, ELF MF)对生物体的可能影响及其可能致癌性受到广泛关注,国际癌症研究组织(IARC)于2001年将其定义为人类可疑致癌源。但对ELF MF的流行病学研究结论并不一致,ELF MF与肿瘤的关系还需要体内外实验证据的支持。肿瘤发生发展是一个复杂的多阶段过程,其中包括浸润和迁入周围组织、血管新生、淋巴细胞的迁移运输等生理活动,这些过程都需要细胞增殖和迁移的参与。而微丝细胞骨架的重组和循环运动则为其提供了动力和物质基础。已经发现,在多种肿瘤细胞中,肌动蛋白微丝被大量修饰或出现过表达现象,并发现与微丝功能相关的一些重要的信号分子活性或协同表达的上调等与肿瘤的发生和转移密切相关。本研究选择在肿瘤发生发展中与细胞增殖、迁移、信号转导均有密切关系的微丝为主要研究对象。首先采用免疫荧光法,使用罗丹明标记的鬼笔环肽(TRITC-Phalloidin)与纤维化肌动蛋白(F-actin)特异性结合,观察不同强度和时间工频磁场辐照对中国仓鼠肺成纤维细胞(Chinese Hamster Lung, CHL)细胞F-actin的影响；第二,扫描电子显微镜(SEM)观察工频磁场辐照对CHL细胞表面形态的影响；第三,采用划痕实验和MTT法观察与肌动蛋白(actin)相关的细胞迁移和增殖的变化；第四,通过免疫印迹法对其可能的信号通路机制进行初步探索。结果显示50 Hz不同强度工频磁场暴露不同时间可以诱导CHL细胞出现不同程度的F-actin骨架重组和边缘化现象,同时导致细胞周围伪足增多,形态扁平。0.4 mT，，24 h暴露可显著增加细胞迁移速率,但本实验条件下ELF MF并不能引起PI3K和磷酸化Akt蛋白表达的显著变化。
[Abstract]:With the development of electric power industry, the possible effect of very low frequency magnetic field (extremely low frequency magnetic fields, ELF MF) on organisms and its possible carcinogenicity have been paid more and more attention. The (IARC) of the International Organization for Research on Cancer (IARC) defined it as a suspected human carcinogenic source in 2001. However, the epidemiological findings of ELF MF do not agree with the relationship between, ELF MF and tumor, and need to be supported by in vivo and in vitro experimental evidence. Tumorigenesis and development is a complex multi-stage process, including infiltration and migration into surrounding tissues, angiogenesis, lymphocyte migration and other physiological activities, all of which require the participation of cell proliferation and migration. The recombination and cyclic movement of the microfilament cytoskeleton provide the power and material basis for the microfilament cytoskeleton. It has been found that actin microfilaments have been extensively modified or overexpressed in various tumor cells. It was also found that some important signal molecules related to the function of microfilaments or the up-regulation of co-expression were closely related to the occurrence and metastasis of tumor. In this study, microfilaments, which are closely related to cell proliferation, migration and signal transduction, were selected as the main research objects. Firstly, the specific binding of Rhodamine labeled TRITC-Phalloidin to fibroactin (F-actin) was used to observe the effect of different intensity and time frequency magnetic field irradiation on (Chinese Hamster Lung, of Chinese hamster lung fibroblasts by immunofluorescence method. CHL) cell F-actin; Secondly, scanning electron microscope (SEM) was used to observe the effect of power frequency magnetic field irradiation on the surface morphology of CHL cells, third, the changes of cell migration and proliferation associated with actin (actin) were observed by scratch test and MTT method. Fourthly, the possible mechanism of signal pathway was preliminarily explored by Western blotting. The results showed that 50 Hz exposure to 50 Hz power frequency magnetic field could induce the recombination and marginalization of F-actin cytoskeleton in CHL cells in different degrees, at the same time, the pseudopodia around the cells increased and the morphology was flat 0.4 mT,. 24 h exposure significantly increased the cell migration rate, but ELF MF did not cause significant changes in the expression of PI3K and phosphorylated Akt.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R35

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