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向“接触抑制”的3T3细胞中添加血清后迅速引发转运变化。

Transport changes rapidly initiated by serum addition to "contact inhibited" 3T3 cells.

作者信息

Cunningham D D, Pardee A B

出版信息

Proc Natl Acad Sci U S A. 1969 Nov;64(3):1049-56. doi: 10.1073/pnas.64.3.1049.

Abstract

Two- to fourfold increases in uridine and phosphate uptake were brought about within 10 to 15 minutes after adding fresh serum to confluent 3T3 cells. The stimulation of transport was specific, since no serum effect was observed for 3-O-methyl-D-glucose or amino acids. The early increase in RNA labeling, previously identified by (14)C-uridine incorporation, could be accounted for by this transport effect. Increased labeling with (32)P-phosphate of at least five phospholipids occurred soon after adding serum. This increase, however, could not be accounted for by increased transport. Both of these results suggest that specific early membrane changes are involved in "contact inhibition."The following results are consistent with this suggestion. Uptake of uridine and phosphate by nonconfluent 3T3 cells was higher than by confluent cells and was only slightly increased by fresh serum. In contrast, uptake of these substrates by Polyoma virus-transformed 3T3 cells, which are not subject to "contact inhibition," was not significantly decreased after the cells became confluent, and serum addition had no effect on transport by either nonconfluent or confluent Polyoma virus-transformed 3T3 cells. Fractionation of serum on Sephadex G-200 demonstrated that the factor which stimulated phosphate transport was different from the previously identified factor which stimulates DNA synthesis by confluent 3T3 cells.

摘要

向汇合的3T3细胞添加新鲜血清后10至15分钟内,尿苷和磷酸盐摄取增加了2至4倍。转运的刺激是特异性的,因为未观察到3-O-甲基-D-葡萄糖或氨基酸有血清效应。先前通过(14)C-尿苷掺入确定的RNA标记早期增加可由这种转运效应来解释。添加血清后不久,至少五种磷脂的(32)P-磷酸盐标记增加。然而,这种增加不能用转运增加来解释。这两个结果都表明,特定的早期膜变化与“接触抑制”有关。以下结果与该建议一致。未汇合的3T3细胞对尿苷和磷酸盐的摄取高于汇合细胞,并且新鲜血清仅使其略有增加。相反,多瘤病毒转化的3T3细胞(不受“接触抑制”)对这些底物的摄取在细胞汇合后没有明显降低,并且添加血清对未汇合或汇合的多瘤病毒转化的3T3细胞的转运均无影响。在Sephadex G-200上对血清进行分级分离表明,刺激磷酸盐转运的因子与先前鉴定的刺激汇合3T3细胞DNA合成的因子不同。

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