细胞种类及孵育时间对血卟啉单甲醚在细胞内分布的影响

Influence of Cell Type and Incubating Time on Intracellular Distribution of Hematoporphrin Monomethyl Ether

目的探讨细胞种类和孵育时间对光敏剂血卟啉单甲醚(HMME)在细胞内分布的影响。方法传代培养小鼠肺内皮细胞(1H11)和A549肺癌细胞,分别与光敏剂HMME孵育2h和12h。应用由荧光显微镜及CCD组成的高分辨率荧光显微成像系统,结合荧光探针标记技术,采用细胞器-细胞荧光强度比值法研究HMME在不同条件下的亚细胞分布情况。结果在2h和12h2个孵育时间点高尔基体的J1/J2值都最高;随着孵育时间延长,A549细胞的4种细胞器的J1/J2值都升高且溶酶体幅度最大,而小鼠肺内皮细胞只有溶酶体的J1/J2值呈升高趋势,高尔基体、内质网和线粒体等细胞器的J1/J2值均呈下降趋势;A549细胞溶酶体的J1/J2值升高的幅度明显高于鼠肺内皮细胞。结论HMME在不同种类细胞内的亚细胞分布是不同的。孵育时间对HMME亚细胞分布的影响显著。随着孵育时间的延长,A549肺癌细胞各细胞器吸收HMME能力逐渐增强,尤以溶酶体显著;小鼠肺内皮细胞中除溶酶体外各细胞器吸收HMME能力逐渐减弱。孵育12h后,A549肺癌细胞各细胞器吸收HMME能力均强于鼠肺内皮细胞。

Objective To discuss the influence of cell type and incubating time on intracellular distribution of hematoporphrin monomethyl ether(HMME).Methods Murine lung endothelial cells and A549 cells were subcultured, then incubated with HMME for 2h and 12h.High-resolution fluorescence microimaging system consisted of fluorescence microscope and CCD and labeling techonology of fluorescence probes were applied and organelle-cell fluorescence intensity ratio analysis was adopted to study subcellular localization of HMME in various situation.Results J_1/J_2 of Golgi was the highest both at 2h and 12h. With incubating time prolonging, J_1/J_2 of A549 cells' four organelles was increasd , and J_1/J_2 of endothelial cells' lysosome was increased, while that of three other organelles including Golgi, ER and mitochondria was decreased; the increase of J_1/J_2 of A549 cells' lysosome was higher than that of endothelial cells'.Conclusions Intracellular distribution of different cell types is different;the influence of incubating time on subcellular distribution of HMME is significant. With incubating time prolonging, four organelles' ability to absorb HMME is enhancing in A549 cells, especially does lysosome; three organelles' ability to absorb HMME is weakening in endothelial cells, except lysosome. After incubating for 12h, A549 cells' four organelles' ability to absorb HMME is more power than that of endothelial cells.