雷帕霉素诱导自噬对结肠癌细胞侵袭、迁移的影响及其可能机制

Effect of autophagy on colon cells' invasion and migration induced by rapamycin and its possible mechanisms

目的观察雷帕霉素(rapamycin,RAPA)诱导结肠癌SW480细胞自噬后对细胞侵袭、迁移能力的影响,并探讨其可能的机制。方法采用免疫荧光观察自噬泡,透射电子显微镜观察细胞超微结构,免疫印记法检测LC3水平,Boyden小室观察细胞的侵袭,划痕实验观察细胞迁移,流式细胞术检测细胞内活性氧(reactive oxygen species,ROS)水平。结果 RAPA处理后结肠癌细胞发生自噬,且自噬水平随处理时间的延长而升高;RAPA诱导自噬组侵袭能力增强(P<0.01),且迁移能力增加(P<0.05);RAPA处理组细胞内ROS水平增高。结论 RAPA可诱导结肠癌细胞发生自噬,使细胞侵袭力增强、迁移力增加,且涉及胞内ROS水平的改变。

Purpose To observe the effects of rapamycin-induced autophagy on the invasion, migration of SW480 colon cancer cells and to investigate the possible mechanism. Methods The autophagy vesicles were detected by monodansyleadaverin （MDC） fluores- cence. The uhrastructure was observed under transmission electron microscope （TEM）. The level of LC3 induced by rapamyein was detected by Western blotting. The invasion of SW480 colon cells was measured by Boyden chamber. Cell wound healing assay was used to observe the migration of SW480 colon cells. The level of ROS was analyzed using flow cytometry. Results The SW480 colon cells were purified with autophagic vesicle under rapamycin environment and autophagy level also increased in a time-dependent manner. The number of SW480 moving into the second floor was increased under the rapamycin environment （ P 〈 0. 01 ）, while the migration was also improved （P 〈 0. 05 ）. The production of ROS increased as well. Conclusions Rapamyein can induce autophagy on SW480 cell line. For SW480 colon cancer cells, the invasion and migration could be enhanced under the autophagy environment induced by ra- pamycin. The level of ROS also changes under this environment.