Metabolism and Cancer Susceptibility Section, Laboratory of Comparative Carcinogenesis, Center for Cancer Research, National Cancer Institute, Frederick, MD 21702-1201, USA.
Carcinogenesis. 2010 Mar;31(3):446-54. doi: 10.1093/carcin/bgp299. Epub 2009 Nov 25.
Epidemiological studies showed that high levels of oxidized low-density lipoproteins (oxLDLs) are associated with increased cancer risk. We examined the direct effect of physiologic concentrations oxLDL on cancer cells. OxLDLs were cytotoxic and activate both apoptosis and autophagy. OxLDLs have ligands for peroxisome proliferator-activated receptor gamma and upregulated proline oxidase (POX) through this nuclear receptor. We identified 7-ketocholesterol (7KC) as a main component responsible for the latter. To elucidate the role of POX in oxLDL-mediated cytotoxicity, we knocked down POX via small interfering RNA and found that this (i) further reduced viability of cancer cells treated with oxLDL; (ii) decreased oxLDL-associated reactive oxygen species generation; (iii) decreased autophagy measured via beclin-1 protein level and light-chain 3 protein (LC3)-I into LC3-II conversion. Using POX-expressing cell model, we established that single POX overexpression was sufficient to activate autophagy. Thus, it led to autophagosomes accumulation and increased conversion of LC3-I into LC3-II. Moreover, beclin-1 gene expression was directly dependent on POX catalytic activity, namely the generation of POX-dependent superoxide. We conclude that POX is critical in the cellular response to the noxious effects of oxLDL by activating protective autophagy.
流行病学研究表明,氧化型低密度脂蛋白(oxLDL)水平升高与癌症风险增加有关。我们研究了生理浓度 oxLDL 对癌细胞的直接作用。oxLDL 具有细胞毒性,并激活细胞凋亡和自噬。oxLDL 是过氧化物酶体增殖物激活受体 γ(peroxisome proliferator-activated receptor gamma,PPARγ)的配体,并通过该核受体上调脯氨酸氧化酶(proline oxidase,POX)。我们确定 7-酮胆固醇(7-ketocholesterol,7KC)是后者的主要成分。为了阐明 POX 在 oxLDL 介导的细胞毒性中的作用,我们通过小干扰 RNA(small interfering RNA,siRNA)敲低 POX,发现这(i)进一步降低了 oxLDL 处理的癌细胞活力;(ii)降低了 oxLDL 相关的活性氧(reactive oxygen species,ROS)生成;(iii)通过 beclin-1 蛋白水平和 LC3-I 到 LC3-II 转化测量的自噬减少。使用表达 POX 的细胞模型,我们证实单 POX 过表达足以激活自噬。因此,它导致自噬体积累,并增加了 LC3-I 到 LC3-II 的转化。此外,beclin-1 基因表达直接依赖于 POX 的催化活性,即 POX 依赖性超氧的产生。我们得出结论,POX 通过激活保护性自噬在细胞对 oxLDL 的有害作用的反应中至关重要。