Tumorigenic and molecular characterization of novel phorbol ester-resistant and -sensitive lines of mice.
Tumorigenic and molecular characterization of novel phorbol ester-resistant and -sensitive lines of mice.
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新型佛波酯抗性和敏感小鼠系的致瘤和分子特征。
DOI：
10.1002/mc.2940010207
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发表时间：
1988
影响因子：
4.6
通讯作者：
Wille,JJ
中科院分区：
文献类型：
--
作者：
Cope,FO;Wagner,FJ;Conway,T;Burns,PD;Johnston,D;Murphy,P;Triggs,G;Wille,JJ
使用雄性和雌性在起始（二甲基苯并[a]蒽；DMBA）、促进（12-O-十四烷酰佛波醇-13-乙酸酯；TPA）挑战、肿瘤发生后育种方案中培育出两个 CD-1 小鼠的近交系。我们的结果表明，佛波酯抗性（PESTI）品系产生肿瘤的速度比 CD-1 亲本品系快 4.1 倍，而佛波酯抗性（PERTI）品系产生肿瘤的速度比 CD-1 亲本品系慢 36 倍。在 PESTI 系中，每只小鼠的平均肿瘤数量在 12 周时达到 27.5 个水平，在 PERTI 系中在 16 周时达到 0.1 个，在 CD-1 系中在 16 周时达到 6.7 个。生化测试表明，PESTI 系对 TPA 具有高基础水平和增强的表皮鸟氨酸脱羧酶 (E.C. 4.1.1.17) 反应，后者是最大剂量下 PERTI 系的九倍。体内表皮细胞蛋白磷酸化的放射自显影分析表明基础蛋白磷酸化谱存在显着差异（高磷酸盐掺入，PERTI，112.7、95.5、64.4、40.8、18.6、17.4 和 12.3 kDa；PESTI，64.4、40.8、31.8 和 12.3 kDa）以及这些谱中 TPA 依赖性变化（与基础水平的差异，PERTI，31.8 和 12.8 kDa；PESTI，139.6、126.3、37.2 和 18.6 kDa）。这些异质谱表明这些蛋白激酶 C 靶底物存在强烈的遗传分离。这种基于皮肤病学的选择和育种过程还导致脑蛋白激酶 C 的 TPA 结合常数 (Kd) 发生共分离遗传变化，而在 PESTI（细胞溶质，0.50 nM；质膜，1.89 nM）和 PERTI（细胞溶质，2.02 nM；质膜，0.51 nM）小鼠中，细胞溶质和质膜部分发生了逆转。此外，仅 PERTI 系小鼠大脑中 TPA 诱导的 43 和 110 kDa 蛋白质体内磷酸化表明，PESTI 系具有高水平的内源磷酸化或这些蛋白质的表达减少。相反，脑胞质蛋白的体外筛选表明，两个细胞系都具有120 kDa的蛋白质，其磷酸化可以由TPA诱导，而PESTI细胞系具有14 kDa的蛋白质，其磷酸化调节控制与PERTI细胞系不同。因此，这些小鼠系提供了用于定义皮肤肿瘤发生过程并可能将启动促进观察和概念扩展到其他组织的实验系统。
Two outbred lines of CD‐1 mice were developed using males and females in an initiation (dimethyl‐benz[a]anthracene; DMBA), promotion (12‐O‐tetradecanoylphorbol‐13‐acetate; TPA) challenge, posttumorigenesis breeding protocol. Our results indicate that the phorbol ester‐resistive (PESTI) line developed tumors at a rate 4.1 times faster than the CD‐1 parental line, while the phorbol ester‐resistant (PERTI) line developed tumors at a rate 36 times slower than the CD‐1 parents. The average number of tumors per mouse reached levels of 27.5 at 12 wk in the PESTI line, 0.1 at 16 wk in the PERTI line, and 6.7 at 16 wk in the CD‐1 line. Biochemical tests showed that the PESTI line had both a high basal level and an enhanced epidermal ornithine decarboxylase (E.C. 4.1.1.17) response to TPA, the latter being nine times that of the PERTI line at their maximum dosages. An autoradiographic analysis of in vivo epidermal cell protein phosphorylation indicated marked differences in basal protein phosphorylation profiles (with high phosphate incorporation, PERTI, 112.7, 95.5, 64.4, 40.8, 18.6, 17.4, and 12.3 kDa; PESTI, 64.4, 40.8, 31.8, and 12.3 kDa) as well as TPA‐dependent changes in these profiles (difference from basal levels, PERTI, 31.8 and 12.8 kDa; PESTI, 139.6, 126.3, 37.2, and 18.6 kDa). These heterogeneous profiles indicate strong genetic segregation of these protein kinase C target substrates. This dermatologically based selection and breeding process also resulted in cosegregating genetic changes in TPA binding constants (Kd) for brain protein kinase C, which were reversed for cytosolic and plasma membrane fractions in the PESTI (cytosolic, 0.50 nM; plasma membrane, 1.89 nM) and PERTI (cytosolic, 2.02 nM; plasma membrane, 0.51 nM) mice. Moreover, TPA‐inducible phosphorylation of 43 and 110 kDa proteins in vivo in the brains of only the PERTI line mice suggests that the PESTI line has either high levels of endogenous phosphorylation or reduced expression of these proteins. In contrast, in vitro screening of brain cytosolic proteins indicated that both lines have a 120 kDa protein whose phosphorylation can be induced by TPA while the PESTI line has a 14 kDa protein whose phosphorylation regulatory controls differ from the PERTI line. Thus, these lines of mice provide experimental systems for defining the skin tumorigenesis process and possibly extending initiationpromotion observations and concepts to other tissues.