摘要
本实验采用体外培养的人胃癌细胞,利用免疫荧光技术和FRAP技术研究细胞骨架和膜表面ConA受体复合物的侧向扩散运动.实验分成5组:①用CB处理细胞5小时;②CB处理5小时后,除去药物,换正常培养液培养1小时;③用秋水仙素处理5小时;④秋水仙素处理细胞5小时后,除去药物,换正常培养液培养2小时;⑤对照组.经CB处理细胞后,胞质微丝减少或消失,但是膜ConA受体复合物的侧向扩散运动增加〔D〕=1.05×10^(-1)cm^2/sec,当除去CB后,微丝重新出现,膜ConA受体复合物的侧向扩散运动又减慢〔D〕=0.81×10^(-1)cm^2/sec,与对照组相似.而无论是用秋水仙素处理细胞,还是撤药后微管收复,都不能看到膜表面ConA受体复合物侧向扩散运动的变化.以上结果表明:影响膜表面ConA受体复合物的侧向扩散运动主要在于微丝的作动.
The purpose of this study is to probe into the cytoskeleton and lateral mobility of Concanavalin A (ConA) receptor complex on the membrane of human gastric cancer cell line MGC 80-3 in vitro by using technique of immunofluorescence and fluorescent recovery after photobleaching (FRAP) .The cytoskeleton was observed with Rhodamine-Phalloidin staining and lateral mobility of ConA receptor complex was determined by the method of FRAP. The experiment was divide into 5 groups. 1) .Cell were treated with cytochalasin B(CB) 1.5 μg/ml for 5 h. 2).Cell were treated with CB, and then cultured again with normal medium for 1 h. 3).Cells were treated with colchicine 0.1 μg/ml for 5h. 4).Cells were treated with colchicine then cultured with normal medium for 2 h. 5) .Control group.We have found that microfilament of cytoplasm was decreased or disappeared and rate of lateral diffusion of ConA r ceptor was increased (D = 1.05 ×10-10cm2/ sec)after the cells were treated with CB. The microfilaments and the rate of lateral diffusion of ConA receptor on cell membrane were recovered (D = 0.81 ? 10-10 cm2 /sec) after CB was removed from culture medium. The rate of lateral diffusion of ConA receptor was not influenced, either the cells were treated by using colchicine or recovered by removing the colchicine from the medium. Results indicate that lateral mobility of cell membrane molecules are influenced by microfilaments but not by microtubules.
出处
《生物物理学报》
CAS
CSCD
北大核心
1991年第4期577-581,共5页
Acta Biophysica Sinica
