BACKGROUND & AIMS: Metallothionein-3 (hMT3) is a structurally unique member of the metallothioneins family of low-mass cysteine-rich proteins. hMT3 has poorly characterized functions, and its importance for hepatocellular carcinoma (HCC) cells has not yet been elucidated. Therefore, we investigated the molecular mechanisms driven by hMT3 with a special emphasis on susceptibility to sorafenib. METHODS: Intrinsically sorafenib-resistant (BCLC-3) and sensitive (Huh7) cells with or without up-regulated hMT3 were examined using cDNA microarray and methods aimed at mitochondrial flux, oxidative status, cell death, and cell cycle. In addition, in ovo/ex ovo chick chorioallantoic membrane (CAM) assays were conducted to determine a role of hMT3 in resistance to sorafenib and associated cancer hallmarks, such as angiogenesis and metastastic spread. Molecular aspects of hMT3-mediated induction of sorafenib-resistant phenotype were delineated using mass-spectrometry-based proteomics. RESULTS: The phenotype of sensitive HCC cells can be remodeled into sorafenib-resistant one via up-regulation of hMT3. hMT3 has a profound effect on mitochondrial respiration, glycolysis, and redox homeostasis. Proteomic analyses revealed a number of hMT3-affected biological pathways, including exocytosis, glycolysis, apoptosis, angiogenesis, and cellular stress, which drive resistance to sorafenib. CONCLUSIONS: hMT3 acts as a multifunctional driver capable of inducing sorafenib-resistant phenotype of HCC cells. Our data suggest that hMT3 and related pathways could serve as possible druggable targets to improve therapeutic outcomes in patients with sorafenib-resistant HCC.

• Klíčová slova
• Hepatocellular carcinoma, Metallothionein-3, Resistance, Sorafenib,
• Publikační typ
• časopisecké články MeSH

Respiratory infections are a real threat for humans, and therefore the pig model is of interest for studies. As one of a case for studies, Actinobacillus pleuropneumoniae (APP) caused infections and still worries many pig breeders around the world. To better understand the influence of pathogenic effect of APP on a respiratory system-lungs and tracheobronchial lymph nodes (TBLN), we aimed to employ matrix-assisted laser desorption/ionization time-of-flight mass spectrometry imaging (MALDI-TOF MSI). In this study, six pigs were intranasally infected by APP and two were used as non-infected control, and 48 cryosections have been obtained. MALDI-TOF MSI and immunohistochemistry (IHC) were used to study spatial distribution of infectious markers, especially interleukins, in cryosections of porcine tissues of lungs (necrotic area, marginal zone) and tracheobronchial lymph nodes (TBLN) from pigs infected by APP. CD163, interleukin 1β (IL‑1β) and a protegrin-4 precursor were successfully detected based on their tryptic fragments. CD163 and IL‑1β were confirmed also by IHC. The protegrin-4 precursor was identified by MALDI-TOF/TOF directly on the tissue cryosections. CD163, IL‑1β and protegrin‑4 precursor were all significantly (p < 0.001) more expressed in necrotic areas of lungs infected by APP than in marginal zone, TBLN and in control lungs.

• Klíčová slova
• Actinobacillus pleuropneumoniae, CD163, MALDI MSI, interleukin 1β, lungs infection, pig model, protegrin‑4 precursor,
• MeSH
• Actinobacillus pleuropneumoniae patogenita   MeSH
• antigen CD163 MeSH
• antigeny diferenciační myelomonocytární metabolismus   MeSH
• biologické markery metabolismus   MeSH
• bronchy metabolismus   MeSH
• CD antigeny metabolismus   MeSH
• infekce bakteriemi rodu Actinobacillus metabolismus   mikrobiologie   MeSH
• infekce dýchací soustavy metabolismus   mikrobiologie   MeSH
• interleukin-1beta metabolismus   MeSH
• kationické antimikrobiální peptidy metabolismus   MeSH
• lymfatické uzliny metabolismus   MeSH
• plíce metabolismus   MeSH
• prasata MeSH
• receptory buněčného povrchu metabolismus   MeSH
• spektrometrie hmotnostní - ionizace laserem za účasti matrice metody   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antigen CD163 MeSH
• antigeny diferenciační myelomonocytární MeSH
• biologické markery MeSH
• CD antigeny MeSH
• interleukin-1beta MeSH
• kationické antimikrobiální peptidy MeSH
• protegrin-4 MeSH Prohlížeč
• receptory buněčného povrchu MeSH