Glycolytic cancer associated fibroblasts promote breast cancer tumor growth, without a measurable increase in angiogenesis Evidence for stromal-epithelial metabolic coupling
Glycolytic cancer associated fibroblasts promote breast cancer tumor growth, without a measurable increase in angiogenesis Evidence for stromal-epithelial metabolic coupling
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DOI：
10.4161/cc.9.12.11989
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发表时间：
2010-06-15
期刊：
影响因子：
4.3
通讯作者：
Lisanti, Michael P.
中科院分区：
文献类型：
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作者：
Migneco, Gemma;Whitaker-Menezes, Diana;Lisanti, Michael P.
此前，我们提出了一个新的模型来理解华宝效应在肿瘤发生和转移中的作用。在这个模型中，间质成纤维细胞将经历有氧糖酵解(也称为Warburg效应)-产生和分泌增加的丙酮酸/乳酸，然后被邻近的上皮癌细胞用作线粒体TCA循环、氧化磷酸化和ATP产生的燃料。为了更直接地测试这一模型，我们在这里使用了一组代谢特征良好的永生化成纤维细胞，这些成纤维细胞在单个基因上存在差异。CL3成纤维细胞向氧化代谢转变，线粒体质量增加。相反，CL4成纤维细胞表现出向有氧糖酵解的转变，并且线粒体质量减少。我们通过无偏倚的蛋白质组学分析验证了CL3和CL4成纤维细胞中的这些差异，显示了CL4成纤维细胞中4种糖酵解酶，即ENO1，ALDOA，LDHA和TPI1的功能上调。正如无偏倚蛋白质组学所见，在CL4成纤维细胞中上调的许多蛋白质，在人类乳腺癌的间质中也转录上调，特别是在容易转移的患者中。重要的是，当CL4成纤维细胞与人乳腺癌细胞(MDA-MB-231)在异种移植模型中共同注射时，肿瘤生长显著增强。CL4成纤维细胞诱导的肿瘤质量增加了4倍，肿瘤体积增加了近8倍，但在肿瘤血管生成方面没有任何可测量的增加。平行地，当单独注射CL3和CL4成纤维细胞时，它们都没有形成肿瘤，没有上皮癌细胞。机制上，在共培养条件下，CL4糖酵解成纤维细胞增加了邻近乳腺癌细胞(相对于CL3细胞)的线粒体活性，这与“反向Warburg效应”是一致的。值得注意的是，对CL4成纤维细胞的Western印迹分析显示，小窝蛋白-1(Cav-1)的蛋白水平显著降低。在人类乳腺癌患者中，间质Cav-1的缺失与早期肿瘤复发、转移、他莫昔芬耐药和不良临床结果的风险增加相关。因此，间质Cav-1的缺失可能是预测乳腺癌患者间质中“反向Warburg效应”的有效标记物。因此，CL4成纤维细胞是模拟癌症相关成纤维细胞“糖酵解表型”的一种新的有吸引力的模型。从糖酵解癌症相关成纤维细胞中提取的营养物质可以通过有效地减少癌细胞对血管血液供应的依赖，在抗血管生成治疗中提供一种逃逸机制来产生耐药性。
Previously, we proposed a new model for understanding the Warburg effect in tumorigenesis and metastasis. In this model, the stromal fibroblasts would undergo aerobic glycolysis (a.k.a., the Warburg effect)-producing and secreting increased pyruvate/lactate that could then be used by adjacent epithelial cancer cells as "fuel" for the mitochondrial TCA cycle, oxidative phosphorylation, and ATP production. to test this model more directly, here we used a matched set of metabolically well-characterized immortalized fibroblasts that differ in a single gene. CL3 fibroblasts show a shift towards oxidative metabolism, and have an increased mitochondrial mass. In contrast, CL4 fibroblasts show a shift towards aerobic glycolysis, and have a reduced mitochondrial mass. We validated these differences in CL3 and CL4 fibroblasts by performing an unbiased proteomics analysis, showing the functional upregulation of 4 glycolytic enzymes, namely ENO1, ALDOA, LDHA and TPI1, in CL4 fibroblasts. Many of the proteins that were upregulated in CL4 fibroblasts, as seen by unbiased proteomics, were also transcriptionally upregulated in the stroma of human breast cancers, especially in the patients that were prone to metastasis. Importantly, when CL4 fibroblasts were co-injected with human breast cancer cells (MDA-MB-231) in a xenograft model, tumor growth was dramatically enhanced. CL4 fibroblasts induced a >4-fold increase in tumor mass, and a near 8-fold increase in tumor volume, without any measurable increases in tumor angiogenesis. In parallel, CL3 and CL4 fibroblasts both failed to form tumors when they were injected alone, without epithelial cancer cells. Mechanistically, under co-culture conditions, CL4 glycolytic fibroblasts increased mitochondrial activity in adjacent breast cancer cells (relative to CL3 cells), consistent with the "Reverse Warburg effect". Notably, Western blot analysis of CL4 fibroblasts revealed a significant reduction in caveolin-1 (Cav-1) protein levels. In human breast cancer patients, a loss of stromal Cav-1 is associated with an increased risk of early tumor recurrence, metastasis, tamoxifen-resistance, and poor clinical outcome. thus, loss of stromal Cav-1 may be an effective marker for predicting the "Reverse Warburg effect" in the stroma of human breast cancer patients. As such, CL4 fibroblasts are a new attractive model for mimicking the "glycolytic phenotype" of cancer-associated fibroblasts. Nutrients derived from glycolytic cancer associated fibroblasts could provide an escape mechanism to confer drug-resistance during anti-angiogenic therapy, by effectively reducing the dependence of cancer cells on a vascular blood supply.