Harshman S, Stoller D, Harshman D L
J Membr Biol. 1979 Oct 15;50(2):165-75. doi: 10.1007/BF01868946.
Contact-inhibited 3T3 mouse fibroblast cells, in contrast to logarithmically growing 3T3 cells and SV-3T3 transformed cells, have increased levels of plasma membrane-bound phosphodiesterase (oligonucleotidase, E.C.3.1.4.19; nucleotide pyrophosphatase, E.C. 3.6.1.9) activity. The increase in enzyme, recorded as increased specific activity, is reversible, as evidenced by the return to normal values following dilution of confluent 3T2 cells and re-initiation of growth. Increased enzyme activity is induced again when the cells regain the confluent state. Transformed SV-3T3 cells can be induced to mimic the contact inhibited state, including increased plasma membrane phosphodiesterase activity, by exposure to a combination of: (i) agents that are known to induce increased intracellular cAMP levels and (ii) additions of purified 3T3 or SV-3T3 plasma membranes. Additions of either alone fails to induce the increase in membrane phosphodiesterase activity, although each alone can significantly suppress cell growth, as measured by incorporation of 3H amino acids. We suggest that the elevation of plasma membrane phosphodiesterase activity may serve as a measure of conversion to the contact-inhibited state in both normal cells and phenotypically reverted transformed cells.
与对数生长期的3T3细胞和SV - 3T3转化细胞相比,接触抑制的3T3小鼠成纤维细胞的质膜结合磷酸二酯酶(寡核苷酸酶,E.C.3.1.4.19;核苷酸焦磷酸酶,E.C. 3.6.1.9)活性水平升高。以比活性增加记录的酶活性增加是可逆的,汇合的3T2细胞稀释并重新开始生长后恢复到正常值就证明了这一点。当细胞重新达到汇合状态时,酶活性再次增加。通过暴露于以下组合,可以诱导转化的SV - 3T3细胞模拟接触抑制状态,包括质膜磷酸二酯酶活性增加:(i)已知可诱导细胞内cAMP水平升高的试剂和(ii)添加纯化的3T3或SV - 3T3质膜。单独添加任何一种都不能诱导膜磷酸二酯酶活性增加,尽管单独使用每种都可以通过3H氨基酸掺入来显著抑制细胞生长。我们认为质膜磷酸二酯酶活性的升高可能作为正常细胞和表型逆转的转化细胞向接触抑制状态转化的一种度量。