Ganglioside-mediated modulation of cell growth, growth factor binding, and receptor phosphorylation.
Ganglioside-mediated modulation of cell growth, growth factor binding, and receptor phosphorylation.
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DOI：
10.1016/s0021-9258(17)39801-0
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发表时间：
1984-06
期刊：
影响因子：
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通讯作者：
E. Bremer;S. Hakomori;D. Bowen-Pope;E. Raines;R. Ross
中科院分区：
文献类型：
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作者：
E. Bremer;S. Hakomori;D. Bowen-Pope;E. Raines;R. Ross
细胞培养中外源性添加的鞘糖脂会缓慢地融入质膜，抑制细胞生长，并改变生长行为（Laine， R. A., and Hakomori， S., 1973）。Biophys。《平民法典》54,1039-1045；Keenan， T. W., Schmid， E., Franke， W. W., and Wiegandt， H. (1975) Exp. Cell, 1992,259 -270)。随着近年来纯化生长因子和无血清培养条件的可用性，我们已经能够在小鼠Swiss 3T3细胞中更详细地研究这一现象，结果如下。1)无血清培养基中的细胞生长（细胞数量增加）受到GM1的特异性抑制，GM3的抑制程度较低，但NeuAcnLc4没有，尽管神经节苷类物质同样很好地融入细胞膜。GM3抑制血小板衍生生长因子(PDGF)-和表皮生长因子刺激的有丝分裂，而GM1只能抑制PDGF-刺激的有丝分裂。NeuAcnLc4对有丝分裂原刺激的胸腺嘧啶掺入无影响。2) 125I-PDGF与细胞的浓度依赖性结合表明，与未添加神经节苷脂的细胞相比，GM1或GM3抑制生长的细胞对PDGF的亲和力增加，而受体总数保持不变。神经节苷脂的加入不影响125I-EGF的结合。3)没有观察到神经节苷脂和生长因子之间的直接相互作用，这是因为含有神经节苷脂的脂质体没有竞争125I生长因子的细胞结合。4) GM1和GM3，而NeuAcnLc4和Gb4都不能抑制PDGF刺激的酪氨酸磷酸化，通过膜法制备一个分子量为170,000的蛋白，该蛋白可能是PDGF受体。因此，膜中神经节苷脂GM1和GM3的水平可能通过影响酪氨酸磷酸化的程度来调节PDGF受体的功能，并可能改变受体对PDGF的亲和力。
Glycosphingolipids added exogenously in cell culture are slowly incorporated into plasma membranes, inhibit cell growth, and modify growth behavior ( Laine , R. A., and Hakomori, S. (1973) Biochem. Biophys. Res. Commun. 54, 1039-1045; Keenan , T. W., Schmid, E., Franke , W. W., and Wiegandt , H. (1975) Exp. Cell Res. 92, 259-270). With the availability of purified growth factors and serum-free culture conditions in recent years, we have been able to examine this phenomenon in mouse Swiss 3T3 cells in greater detail with the following results. 1) Cell growth (cell number increase) in serum-free medium was specifically inhibited by the presence of GM1 and to a lesser extent by GM3, but not by NeuAcnLc4 , although the gangliosides were incorporated equally well into cell membranes. GM3 inhibited both platelet-derived growth factor (PDGF)- and epidermal growth factor-stimulated mitogenesis determined by thymidine incorporation, while GM1 could only inhibit PDGF-stimulated mitogenesis. NeuAcnLc4 had no effect on mitogen-stimulated thymidine incorporation. 2) The concentration-dependent binding of 125I-PDGF binding to cells indicated that cells whose growth was inhibited by GM1 or GM3 showed an increased affinity for PDGF as compared to cells grown without addition of ganglioside, while the total number of receptors stayed the same. Addition of ganglioside did not affect the binding of 125I-EGF. 3) No direct interaction was observed between gangliosides and growth factors as evidenced by the lack of competition by ganglioside-containing liposomes for cellular binding of 125I growth factors. 4) GM1 and GM3, but neither NeuAcnLc4 nor Gb4 , inhibited the PDGF-stimulated tyrosine phosphorylation by membrane preparations of a 170,000 molecular weight protein, which is probably the PDGF receptor. Thus, the level of gangliosides GM1 and GM3 in membranes may modulate PDGF receptor function by affecting the degree of tyrosine phosphorylation and may alter the affinity of the receptor for PDGF.