Kato G, Wakabayashi K
Department of Biochemistry, Yamanashi Medical College, Japan.
In Vitro Cell Dev Biol. 1988 Apr;24(4):274-80. doi: 10.1007/BF02628827.
We have studied the responses of three human retinoblastoma cell lines (GM1232, Y79, and WERI-Rb1) to substratum-bound laminin (LN), fibronectin (FN), or collagen (CN) in serum-containing medium. About 95% of the cells attached to poly-D-lysine (PN)-pretreated plastic surfaces either unbound or protein-bound within 1 h after plating. With PN-bound LN, GM1232 cells showed an outgrowth of processes and cell spread within 1 d, but very little was seen on PN-bound FN, CN, or unbound PN even by Day 4. Both the percentage of cells with processes and the number of processes/cell were dependent on the amount of LN bound to the surfaces at Day 4. On LN surfaces without PN pretreatment, by Day 2 most cells had formed floating aggregates and were not attached to the surfaces. By Day 4 only a part of the peripheral cells of attached aggregates displayed process outgrowth and cell spreading. Dibutyryl cyclic AMP (dbcAMP) maintained these effects of PN-bound LN, and promoted process branching, cell spreading, and elongation with concomitant inhibition of cell growth. The percentage of cells with processes was 83%. Y79 and WERI-Rb1 cells, passed for several years, showed little and weak response to PN-bound LN either in the absence or presence of dbcAMP. These results indicate that the morphologic differentiation of GM1232 cells is elicited specifically by PN-bound LN, and that dbcAMP maintains and promotes this differentiated status.
我们研究了三种人视网膜母细胞瘤细胞系(GM1232、Y79和WERI-Rb1)在含血清培养基中对基质结合层粘连蛋白(LN)、纤连蛋白(FN)或胶原蛋白(CN)的反应。接种后1小时内,约95%的细胞附着在聚-D-赖氨酸(PN)预处理的塑料表面,这些表面要么未结合蛋白质,要么结合了蛋白质。对于PN结合的LN,GM1232细胞在1天内出现突起生长和细胞铺展,但即使到第4天,在PN结合的FN、CN或未结合PN的表面上也很少见到这种情况。到第4天,有突起的细胞百分比和每个细胞的突起数量都取决于表面结合的LN量。在未经PN预处理的LN表面,到第2天,大多数细胞形成了漂浮聚集体,未附着在表面。到第4天,只有附着聚集体周边的部分细胞显示出突起生长和细胞铺展。二丁酰环磷腺苷(dbcAMP)维持了PN结合的LN的这些作用,并促进了突起分支、细胞铺展和伸长,同时抑制细胞生长。有突起的细胞百分比为83%。经过数年传代的Y79和WERI-Rb1细胞,无论在有无dbcAMP的情况下,对PN结合的LN的反应都很小且微弱。这些结果表明,GM1232细胞的形态分化是由PN结合的LN特异性引发的,并且dbcAMP维持并促进了这种分化状态。