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Abstract：
BACKGROUND: Metastasis is the leading cause of cancer-related death in non-small cell lung cancer (NSCLC) patients. We previously showed that low HERC5 expression predicts early tumor dissemination and a dismal prognosis in NSCLC patients. Here, we performed functional studies to unravel the mechanism underlying the \"metastasis-suppressor\" effect of HERC5, with a focus on mitochondrial metabolism pathways.
METHODS: We assessed cell proliferation, colony formation potential, anchorage-independent growth, migration, and wound healing in NSCLC cell line models with HERC5 overexpression (OE) or knockout (KO). To study early tumor cell dissemination, we used these cell line models in zebrafish experiments and performed intracardial injections in nude mice. Mass spectrometry (MS) was used to analyze protein changes in whole-cell extracts. Furthermore, electron microscopy (EM) imaging, cellular respiration, glycolytic activity, and lactate production were used to investigate the relationships with mitochondrial energy metabolism pathways.
RESULTS: Using different in vitro NSCLC cell line models, we showed that NSCLC cells with low HERC5 expression had increased malignant and invasive properties. Furthermore, two different in vivo models in zebrafish and a xenograft mouse model showed increased dissemination and metastasis formation (in particular in the brain). Functional enrichment clustering of MS data revealed an increase in mitochondrial proteins in vitro when HERC5 levels were high. Loss of HERC5 leads to an increased Warburg effect, leading to improved adaptation and survival under prolonged inhibition of oxidative phosphorylation.
CONCLUSIONS: Taken together, these results indicate that low HERC5 expression increases the metastatic potential of NSCLC in vitro and in vivo. Furthermore, HERC5-induced proteomic changes influence mitochondrial pathways, ultimately leading to alterations in energy metabolism and demonstrating its role as a new potential metastasis suppressor gene.
METHODS: We assessed cell proliferation, colony formation potential, anchorage-independent growth, migration, and wound healing in NSCLC cell line models with HERC5 overexpression (OE) or knockout (KO). To study early tumor cell dissemination, we used these cell line models in zebrafish experiments and performed intracardial injections in nude mice. Mass spectrometry (MS) was used to analyze protein changes in whole-cell extracts. Furthermore, electron microscopy (EM) imaging, cellular respiration, glycolytic activity, and lactate production were used to investigate the relationships with mitochondrial energy metabolism pathways.
RESULTS: Using different in vitro NSCLC cell line models, we showed that NSCLC cells with low HERC5 expression had increased malignant and invasive properties. Furthermore, two different in vivo models in zebrafish and a xenograft mouse model showed increased dissemination and metastasis formation (in particular in the brain). Functional enrichment clustering of MS data revealed an increase in mitochondrial proteins in vitro when HERC5 levels were high. Loss of HERC5 leads to an increased Warburg effect, leading to improved adaptation and survival under prolonged inhibition of oxidative phosphorylation.
CONCLUSIONS: Taken together, these results indicate that low HERC5 expression increases the metastatic potential of NSCLC in vitro and in vivo. Furthermore, HERC5-induced proteomic changes influence mitochondrial pathways, ultimately leading to alterations in energy metabolism and demonstrating its role as a new potential metastasis suppressor gene.
摘要：
背景：转移是非小细胞肺癌（NSCLC）患者癌症相关死亡的主要原因。我们先前表明，低HERC5表达预测NSCLC患者的早期肿瘤扩散和预后不良。这里,我们进行了功能研究,以阐明HERC5"转移抑制"效应的潜在机制,重点是线粒体代谢途径.
方法：我们评估了细胞增殖，菌落形成潜力，锚定独立生长，迁移,和具有HERC5过表达(OE)或敲除(KO)的NSCLC细胞系模型中的伤口愈合。为了研究早期肿瘤细胞播散，我们在斑马鱼实验中使用了这些细胞系模型，并在裸鼠中进行了心内注射。质谱(MS)用于分析全细胞提取物中的蛋白质变化。此外，电子显微镜（EM）成像，细胞呼吸，糖酵解活性,和乳酸的产生被用来研究与线粒体能量代谢途径的关系。
结果：使用不同的体外NSCLC细胞系模型，我们发现HERC5低表达的NSCLC细胞的恶性和侵袭性增加.此外，两种不同的斑马鱼体内模型和异种移植小鼠模型显示扩散和转移形成增加（特别是在大脑中）。MS数据的功能富集聚类显示，当HERC5水平高时，线粒体蛋白在体外增加。HERC5的丢失导致Warburg效应增加，导致在长期抑制氧化磷酸化的情况下改善适应性和存活。
结论：综合来看，这些结果表明低HERC5表达增加了NSCLC的体内外转移潜能.此外，HERC5诱导的蛋白质组变化影响线粒体途径，最终导致能量代谢的改变，并证明其作为新的潜在转移抑制基因的作用。
方法：我们评估了细胞增殖，菌落形成潜力，锚定独立生长，迁移,和具有HERC5过表达(OE)或敲除(KO)的NSCLC细胞系模型中的伤口愈合。为了研究早期肿瘤细胞播散，我们在斑马鱼实验中使用了这些细胞系模型，并在裸鼠中进行了心内注射。质谱(MS)用于分析全细胞提取物中的蛋白质变化。此外，电子显微镜（EM）成像，细胞呼吸，糖酵解活性,和乳酸的产生被用来研究与线粒体能量代谢途径的关系。
结果：使用不同的体外NSCLC细胞系模型，我们发现HERC5低表达的NSCLC细胞的恶性和侵袭性增加.此外，两种不同的斑马鱼体内模型和异种移植小鼠模型显示扩散和转移形成增加（特别是在大脑中）。MS数据的功能富集聚类显示，当HERC5水平高时，线粒体蛋白在体外增加。HERC5的丢失导致Warburg效应增加，导致在长期抑制氧化磷酸化的情况下改善适应性和存活。
结论：综合来看，这些结果表明低HERC5表达增加了NSCLC的体内外转移潜能.此外，HERC5诱导的蛋白质组变化影响线粒体途径，最终导致能量代谢的改变，并证明其作为新的潜在转移抑制基因的作用。
