Persistent induction of cyclooxygenase in p60v-src-transformed 3T3 fibroblasts.
Persistent induction of cyclooxygenase in p60v-src-transformed 3T3 fibroblasts.
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DOI：
10.1073/pnas.87.9.3373
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发表时间：
1990-05
影响因子：
11.1
通讯作者：
Jiawen Han;H. Sadowski;D. Young;I. Macara
中科院分区：
文献类型：
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作者：
Jiawen Han;H. Sadowski;D. Young;I. Macara
用温度敏感性劳斯肉瘤病毒株LA 90感染的BALB/c 3 T3细胞系已被用于研究基因表达（蛋白质合成）的早期p60 v-src依赖性变化。巨型双向电泳可从[35 S]甲硫氨酸标记的细胞裂解物中分离出3000多个多肽，用于检测p60 v-src在35 ℃活化后p72-74（72-74 kDa）双联体（pI 7.5）的诱导。抗环氧合酶（前列腺素合酶或前列腺素内过氧化物合酶）的抗血清特异性免疫沉淀p72-74双联体。血小板衍生生长因子和佛波醇12-肉豆蔻酸酯13-乙酸酯也诱导p72-74双联体，并且在野生型src转化的NIH 3 T3细胞系中升高。p60 v-src的激活引起p72-74的持续增加，而生长因子的作用是短暂的。这些不同的诱导动力学是由环氧合酶活性的变化来解释的。蛋白激酶C的下调抑制随后的诱导环氧合酶的佛波酯肉豆蔻酸酯醋酸酯，但不阻止诱导p60 v-src。糖皮质激素激动剂地塞米松抑制p60 v-src对环氧合酶的诱导。虽然这种酶的诱导可能不直接参与转化，但数据支持这样的观点，即致癌转化可能不是由转化特异性基因的表达引起的，而是由通常在细胞周期G 0期通过期间仅瞬时诱导的基因表达的持续变化引起的。
A BALB/c 3T3 cell line infected with the temperature-sensitive Rous sarcoma virus strain LA90 has been used to investigate early, p60v-src-dependent changes in gene expression (protein synthesis). Giant two-dimensional electrophoresis, which can resolve greater than 3000 polypeptides from [35S]methionine-labeled cell lysates, was used to detect the induction of a p72-74 (72-74 kDa) doublet (pI 7.5) after activation of p60v-src at 35 degrees C. Antiserum against cyclooxygenase (prostaglandin synthase or prostaglandin endoperoxide synthase) specifically immunoprecipitated the p72-74 doublet. The p72-74 doublet was also induced by platelet-derived growth factor and by phorbol 12-myristate 13-acetate and was elevated in an NIH 3T3 cell line transformed by wild-type src. Activation of p60v-src caused a persistent increase in p72-74, whereas the effect of the growth factor was transient. These dissimilar kinetics of induction were paralleled by changes in cyclooxygenase activity. Down-regulation of protein kinase C inhibited subsequent induction of cyclooxygenase by phorbol myristate acetate but did not block induction by p60v-src. The glucocorticoid agonist dexamethasone inhibited induction of cyclooxygenase by p60v-src. Although induction of this enzyme may not be directly involved in transformation, the data support the view that oncogenic transformation may result, not from expression of transformation-specific genes, but from persistent changes in the expression of genes normally induced only transiently during passage from the G0 stage of the cell cycle.