Goshima K, Masuda A, Owaribe K
J Cell Biol. 1984 Mar;98(3):801-9. doi: 10.1083/jcb.98.3.801.
Insulin induced the formation of ruffling membranes in cultured KB cells (a cell strain derived from human epidermoid carcinoma) within 1-2 min after its addition. The ruffled regions were stained strongly with antibody to actin but not that to tubulin. Pretreatment of KB cells with agents disrupting microfilaments (cytochalasins), but not with those disrupting microtubules (colcemid, nocodazole, and colchicine) completely inhibited the formation of ruffling membranes. Pretreatment of KB cells with dibutyryl cyclic AMP, but not with dibutyryl cyclic GMP, also inhibited the formation of ruffling membranes. Addition of insulin enhanced Na+-dependent uptake of a system A amino acid (alpha-amino isobutyric acid; AIB) by the cells within 5 min after the addition, and decreased the cyclic AMP content of the cells. Treatments that inhibited insulin-induced formation of ruffling membranes of KB cells also inhibited insulin-induced enhancement of their AIB uptake. From these observations, the mechanism of insulin-induced formation of ruffling membranes and its close correlation with AIB transport are discussed.
胰岛素在添加到培养的KB细胞(一种源自人表皮样癌的细胞系)后1 - 2分钟内诱导了褶皱膜的形成。褶皱区域用抗肌动蛋白抗体染色强烈,但用抗微管蛋白抗体染色不强烈。用破坏微丝的试剂(细胞松弛素)预处理KB细胞,但不用破坏微管的试剂(秋水仙酰胺、诺考达唑和秋水仙碱)预处理,完全抑制了褶皱膜的形成。用二丁酰环磷酸腺苷预处理KB细胞,但不用二丁酰环磷酸鸟苷预处理,也抑制了褶皱膜的形成。添加胰岛素后5分钟内增强了细胞对系统A氨基酸(α-氨基异丁酸;AIB)的钠依赖性摄取,并降低了细胞的环磷酸腺苷含量。抑制胰岛素诱导的KB细胞褶皱膜形成的处理也抑制了胰岛素诱导的其AIB摄取增强。基于这些观察结果,讨论了胰岛素诱导褶皱膜形成的机制及其与AIB转运的密切关系。