Differential gene expression in p53-mediated apoptosis-resistant vs. apoptosis-sensitive tumor cell lines
Differential gene expression in p53-mediated apoptosis-resistant vs. apoptosis-sensitive tumor cell lines
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DOI：
10.1073/pnas.230445997
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发表时间：
2000-11-21
影响因子：
11.1
通讯作者：
Davis, GE
中科院分区：
文献类型：
--
作者：
Maxwell, SA;Davis, GE
用p53重组腺病毒（Ad5CMV-p53）在ECV-304膀胱癌细胞株中诱导野生型p53，导致几乎所有细胞广泛凋亡和最终死亡。为了确定这些转化细胞中p53介导的凋亡的重要分子事件，我们通过Ad5CMV-p53反复感染，选择对p53有抗性的ECV-304细胞，通过逆转录pcr、Northern blotting和DNA微阵列分析，比较了5330个基因在p53抗性（DECV）和p53敏感的ECV-304细胞中的表达。在两种p53感染细胞系中，480个基因的表达差异为2倍或更多。许多p53的潜在靶点被发现在细胞周期调节、DNA修复、氧化还原控制、细胞粘附、细胞凋亡和分化中发挥作用。脯氨酸氧化酶，一种参与脯氨酸/吡啶-5-羧酸氧化还原循环的线粒体酶，在ECV细胞中被p53上调，而在DECV细胞中没有上调。脯氨酸氧化酶产生的脯氨酸衍生代谢物pyroline -5-carboxylate （P5C）抑制ECV-304和DECV细胞的增殖和存活，并诱导两种细胞系的凋亡。一种在线粒体氨基端标记绿色荧光蛋白的重组脯氨酸氧化酶蛋白诱导p53缺失的H1299非小细胞肺癌细胞凋亡。结果直接提示脯氨酸氧化酶和脯氨酸/P5C通路参与p53诱导的生长抑制和凋亡。
Induction of wild-type p53 in the ECV-304 bladder carcinoma cell line by infection with a p53 recombinant adenovirus (Ad5CMV-p53) resulted in extensive apoptosis and eventual death of nearly all of the cells. As a strategy to determine the molecular events important to p53-mediated apoptosis in these transformed cells, ECV-304 cells were selected for resistance to p53 by repeated infections with Ad5CMV-p53, We compared the expression of 5,730 genes in p53-resistant (DECV) and p53-sensitive ECV-304 cells by reverse transcription-PCR, Northern blotting, and DNA microarray analysis. The expression of 480 genes differed by 2-fold or more between the two p53-infected cell lines. A number of potential targets for p53 were identified that play roles in cell cycle regulation, DNA repair, redox control, cell adhesion, apoptosis, and differentiation. Proline oxidase, a mitochondrial enzyme involved in the proline/pyrroline-5-carboxylate redox cycle, was up-regulated by p53 in ECV but not in DECV cells. Pyrroline-5-carboxylate (P5C), a proline-derived metabolite generated by proline oxidase, inhibited the proliferation and survival of ECV-304 and DECV cells and induced apoptosis in both cell lines. A recombinant proline oxidase protein tagged with a green fluorescent protein at the amino terminus localized to mitochondria and induced apoptosis in p53-null H1299 non-small cell lung carcinoma cells. The results directly implicate proline oxidase and the proline/P5C pathway in p53-induced growth suppression and apoptosis.