Additional file 1 of Mir-675-5p supports hypoxia-induced drug resistance in colorectal cancer cells

Additional file 1.

Hypoxia-Induced miR-675-5p Supports β-Catenin Nuclear Localization by Regulating GSK3-β  Activity in Colorectal Cancer Cell Lines

The reduction of oxygen partial pressure in growing tumors triggers numerous survival strategies driven by the transcription factor complex HIF1 (Hypoxia Inducible Factor-1). Recent evidence revealed that HIF1 promotes rapid and effective phenotypic changes through the induction of non-coding RNAs, whose contribution has not yet been fully described. Here we investigated the role of the hypoxia-induced, long non-coding RNA H19 (lncH19) and its intragenic miRNA (miR-675-5p) into HIF1-Wnt crosstalk. During hypoxic stimulation, colorectal cancer cell lines up-regulated the levels of both the lncH19 and its intragenic miR-675-5p. Loss of expression experiments revealed that miR-675-5p inhibitio…

Mouse mesoangioblast behaviour when subjected to cellular stress

Mir-675-5p supports hypoxia-induced drug resistance in colorectal cancer cells.

Abstract Background The uncontrolled proliferation of cancer cells determines hypoxic conditions within the neoplastic mass with consequent activation of specific molecular pathways that allow cells to survive despite oxygen deprivation. The same molecular pathways are often the cause of chemoresistance. This study aims to investigate the role of the hypoxia-induced miR-675-5p in 5-Fluorouracil (5-FU) resistance on colorectal cancer (CRC) cells. Methods CRC cell lines were treated with 5-Fu and incubated in normoxic or hypoxic conditions; cell viability has been evaluated by MTT assay. MiR-675-5p levels were analysed by RT-PCR and loss and gain expression of the miRNA has been obtained by t…

Analisi dei fattori che rendono le cellule di adenocarcinoma colorettale resistenti o suscettibili all’azione di chemioterapici

Extracellular Vesicles: The New Frontier of Stem Cell Regenerative Medicine?

Regenerative medicine aims to repair damaged or missing cells, tissues or organs for the treatment of various diseases, poorly managed with conventional drugs and medical procedures. To date there are different approaches to obtain these results. Multimodal regenerative methods include transplant of healthy organs, tissues, or cells, body stimulation to activate a self healing response in damaged tissues, as well as the combined use of cells and bio-degradable scaffold to obtain functional tissues. Certainly, stem cells and derived products are promising tools in regenerative medicine due to their ability to induce de novo tissue formation and/or promote tissue and organ repair and regenera…

Effetti dell'espressione di K-RasG12V e K-RasG13D in cellule di adenocarcinoma colorettale HT29

RAS è una piccola proteina di 21kDa che si trova frequentemente mutata nei tumori. La famiglia dei geni ras consta di tre principali protoncogeni chiamati H-, K- e N-Ras. Le tre isoforme di Ras regolano la proliferazione, il differenziamento e la morte cellulare mediante l’attivazione di diversi pathways di trasduzione del segnale fra cui la cascata delle MAP chinasi e il pathway di PI3K/AKT. Le diverse isoforme di Ras attivano tutte gli stessi pathways, ma con diversa efficienza, e ciò potrebbe essere, almeno in parte, una conseguenza delle loro differenti modifiche post-traduzionali che determinano la localizzazione in specifici microdomini della membrana plasmatica. Diversi studi hanno d…

Another function of Hsp70 in mesoangioblast stem cells

In recent years, it has been demonstrated that Hsp70 is released in the extracellular space under normal cell culture conditions, and this release is mediated through exosomes. We have demonstrated that A6 cells, a clone of mouse mesoangioblasts, produce and release in the extracellular space membrane vesicles, independently of culture growth conditions. These vesicles contains both structural proteins and biological active molecules, such as FGF-2 and the metalloproteinases MMP 2 and 9. We have also demonstrated that A6 vesicles contain HspP70 and its release is highly regulated. Some of the intracellular Hsp70 is localized on lipid rafts and its concentration in insoluble fraction increas…

Paracrine roles of extracellular vesicles released by mouse mesoangioblasts

Extracellular vesicles (EV) represent an important mediator of cell-to-cell communication and are involved in both autocrine and paracrine signaling, with a critical role in a number of physiological and pathological conditions.1 The bioactive molecules contained within EV simultaneously activate several different pathways resulting in the synergistic stimulation of target cells. The discovery and characterization of EV have added a novel understanding to regenerative medicine, namely the finding that stem cells are an abundant source of EV.1-2 A6 mouse mesoangioblasts, vessel-associated multipotent progenitor stem cells that are capable of differentiating into different mesodermal cell typ…

Codon 12 and codon 13 mutations in K-RAS differentially affect therapies response of colorectal carcinoma cells

Oxidative stress preconditioning of mouse perivascular myogenic progenitors selects a subpopulation of cells with a distinct survival advantage in vitro and in vivo

AbstractCell engraftment, survival and integration during transplantation procedures represent the crux of cell-based therapies. Thus, there have been many studies focused on improving cell viability upon implantation. We used severe oxidative stress to select for a mouse mesoangioblast subpopulation in vitro and found that this subpopulation retained self-renewal and myogenic differentiation capacities while notably enhancing cell survival, proliferation and migration relative to unselected cells. Additionally, this subpopulation of cells presented different resistance and recovery properties upon oxidative stress treatment, demonstrating select advantages over parental mesoangioblasts in …

Effects of Flavonoids on Cancer, Cardiovascular and Neurodegenerative Diseases: Role of NF-κB Signaling Pathway

Flavonoids are polyphenolic phytochemical compounds found in many plants, fruits, vegetables, and leaves. They have a multitude of medicinal applications due to their anti-inflammatory, antioxidative, antiviral, and anticarcinogenic properties. Furthermore, they also have neuroprotective and cardioprotective effects. Their biological properties depend on the chemical structure of flavonoids, their mechanism of action, and their bioavailability. The beneficial effects of flavonoids have been proven for a variety of diseases. In the last few years, it is demonstrated that the effects of flavonoids are mediated by inhibiting the NF-κB (Nuclear Factor-κB) pathway. In this review, we have summar…

Role of Hsp70 in Multiple Sclerosis: An Overview

For many years heat shock protein 70 (Hsp70) was considered exclusively an intracellular chaperone contributing to protein proteostasis and in apoptotic pathway block. Lately it has been demonstrated that Hsp70 is actively released in the extracellular environment, thereby promoting the activation of the immune system by stimulating innate and adaptive responses through the activation of APCs. Its expression in the nervous system is induced in a variety of pathological conditions. Emerging evidences displayed that Hsp70 is a critical regulator in normal neural cells. Multiple sclerosis (MS) is an autoimmune disease of the central nervous system (CNS) directed against myelin antigens. In thi…

Analisi degli effetti del Cetuximab in linee cellulari di adenocarcinoma colorettale

Il carcinoma del colon-retto (CRC) è una malattia eterogenea, che si sviluppa in seguito a numerose alterazioni genetiche ed epigenetiche. Una famiglia genica che si trova frequentemente mutata nei tumori è quella di ras che consta di tre principali protoncogeni (H-, K- e N-Ras) localizzati su cromosomi differenti e codificanti proteine G del peso di 21 KDa. Il 50% dei casi di CRC presenta mutazioni puntiformi missense a carico del gene KRAS, il 90% delle quali si verifica a livello dei codoni 12 e 13 e rende la proteina costitutivamente attiva. Diversi studi dimostrano come mutazioni di differenti codoni di K-Ras potrebbero avere diverse conseguenze biologiche e determinare una diversa ris…

Differences in intercellular communication during clinical relapse and gadolinium-enhanced MRI in patients with relapsing remitting multiple sclerosis: A study of the composition of extracellular vesicles in cerebrospinal fluid

This study was designed based on the hypothesis that changes in both the levels and surface marker expression of extracellular vesicles (EVs) isolated from the cerebrospinal fluid (CSF) may be associated with the clinical form, disease activity, and severity of multiple sclerosis (MS). The analyzes were performed on subjects affected by MS or other neurological disorders. EVs, which were isolated by ultracentrifugation of CSF samples, were characterized by flow cytometry. A panel of fluorescent antibodies was used to identify the EV origin: CD4, CCR3, CCR5, CD19, and CD200, as well as isolectin IB4. The Mann–Whitney U-test and Kruskal–Wallis test were used for statistical analyzes. EVs isol…

Nobiletin and xanthohumol counteract the TNFα-mediated activation of endothelial cells through the inhibition of the NF-κB signaling pathway.

Angiogenesis, a process characterized by the formation of new blood vessels from pre-existing ones, is a crucial step in tumor growth and dissemination. Given the ability of tumors to interfere with multiple or different molecular pathways to promote angiogenesis, there is an increasing need to therapeutically block tumor progression by targeting multiple antiangiogenic pathways. Natural polyphenols present health-protective properties, which are likely attributed to their ability to activate multiple pathways involved in inflammation, carcinogenesis, and angiogenesis. Recently, increased attention has been addressed to the ability of flavonoids, the most abundant polyphenols in the diet, t…

Distinct biological effects are observed in HT-29 colorectal carcinoma cells induced to express K-RASG12V or K-RASG13D

Paracrine effect of membrane vesicles released by mouse mesoangioblast stem cells

Introduction: Mouse mesoangioblasts are vessel-associated multipotent progenitor stem cells, which are able to differentiate into different mesodermal cell types. In our previous paper, we have demonstrated that mesoangioblasts are able to shed in the extracellular environment membrane vesicles (EVs), which contain both structural proteins and biological factors such as FGF2 and the two gelatinases MMP2/9. We investigated whether these EV interact in a paracrine way with other cell types different from mesoangioblasts, and eventually the effects of this interaction. Methods: Mesoangioblast EVs were collected from conditioned media by ultracentrifugation. Total mRNAs from mesoangioblasts and…

Effects of H2O2 on mesoangioblast stem cells: survival and cell death

Hypoxia‐induced non‐coding rnas controlling cell viability in cancer

Hypoxia, a characteristic of the tumour microenvironment, plays a crucial role in cancer progression and therapeutic response. The hypoxia-inducible factors (HIF-1α, HIF-2α, and HIF-3α), are the master regulators in response to low oxygen partial pressure, modulating hypoxic gene expression and signalling transduction pathways. HIFs’ activation is sufficient to change the cell phenotype at multiple levels, by modulating several biological activities from metabolism to the cell cycle and providing the cell with new characteristics that make it more aggressive. In the past few decades, growing numbers of studies have revealed the importance of non-coding RNAs (ncRNAs) as molecular mediators i…

Mesoangioblast behaviour in oxidative stress condition

Mesenchymal and Induced Pluripotent Stem Cells-Derived Extracellular Vesicles: The New Frontier for Regenerative Medicine?

Regenerative medicine aims to repair damaged, tissues or organs for the treatment of various diseases, which have been poorly managed with conventional drugs and medical procedures. To date, multimodal regenerative methods include transplant of healthy organs, tissues, or cells, body stimulation to activate a self-healing response in damaged tissues, as well as the combined use of cells and bio-degradable scaffold to obtain functional tissues. Certainly, stem cells are promising tools in regenerative medicine due to their ability to induce de novo tissue formation and/or promote organ repair and regeneration. Currently, several studies have shown that the beneficial stem cell effects, espec…

Extracellular Hsp70 Enhances Mesoangioblast Migration via an Autocrine Signaling Pathway

Mouse mesoangioblasts are vessel-associated progenitor stem cells endowed with the ability of multipotent mesoderm differentiation. Therefore, they represent a promising tool in the regeneration of injured tissues. Several studies have demonstrated that homing of mesoangioblasts into blood and injured tissues are mainly controlled by cytokines/chemokines and other inflammatory factors. However, little is known about the molecular mechanisms regulating their ability to traverse the extracellular matrix (ECM). Here, we demonstrate that membrane vesicles released by mesoangioblasts contain Hsp70, and that the released Hsp70 is able to interact by an autocrine mechanism with Toll-like receptor …

Extracellular Hsp70 Enhances Mesoangioblast Migration via an Autocrine Signaling Pathway

Mouse mesoangioblasts are vessel-associated progenitor stem cells endowed with the ability of multipotent mesoderm differentiation. Therefore, they represent a promising tool in the regeneration of injured tissues. Several studies have demonstrated that homing of mesoangioblasts into blood and injured tissues are mainly controlled by cytokines/chemokines and other inflammatory factors. However, little is known about the molecular mechanisms regulating their ability to traverse the extracellular matrix (ECM). Here, we demonstrate that membrane vesicles released by mesoangioblasts contain Hsp70, and that the released Hsp70 is able to interact by an autocrine mechanism with Toll-like receptor …

H2O2 resistant mesoangioblast clone isolation with a distinct survival advantage in vitro and in vivo

The release of molecules from damaged tissues stimulates both resident and circulating stem cells to initiate a tissue repair programme. 1 However, during transplantation procedures the therapeutic efficacy of stem cells is compromised by reduced homing capability towards the target site.2,3 Furthermore, cell survival is very low and many studies focused on improving cell viability upon implantation. In this study, we performed in vitrosevere oxidative stress to select some more resistant mouse mesoangioblasts, vessel-associated progenitor stem cells endowed with the ability of multipotent mesoderm differentiation. We found that the selected subpopulation retains selfrenewal and myogenic di…

Rapamycin-Loaded Polymeric Nanoparticles as an Advanced Formulation for Macrophage Targeting in Atherosclerosis

Recently, rapamycin (Rapa) represents a potential drug treatment to induce regression of atherosclerotic plaques

Extracellular vesicles in multiple sclerosis as possible biomarkers: Dream or reality?

Extracellular vesicles are recently described as specialized structures for intercellular communication. Their role in the central nervous system was diffusely studied in both physiological and pathological condition. In particular, an increased extracellular vesicle number was detected in several autoimmune diseases, including multiple sclerosis, a chronic autoimmune, inflammatory, demyelinating and neurodegenerative disease. This chapter summarizes the available information on the involvement of the extracellular vesicles in multiple sclerosis pathogenesis and their possible use as biomarker of therapy efficacy.

Analysis of the mechanisms through which K-RASG12V and K-RASG13D regulate the proliferation and cell death in cells HT-29

Codon 12 and codon 13 mutations in K-RAS differentially affect colorectal carcinoma cells

Molecular Mediators of RNA Loading into Extracellular Vesicles

In the last decade, an increasing number of studies have demonstrated that non-coding RNA (ncRNAs) cooperate in the gene regulatory networks with other biomolecules, including coding RNAs, DNAs and proteins. Among them, microRNAs (miRNAs), long non-coding RNAs (lncRNAs) and circular RNAs (circRNAs) are involved in transcriptional and translation regulation at different levels. Intriguingly, ncRNAs can be packed in vesicles, released in the extracellular space, and finally internalized by receiving cells, thus affecting gene expression also at distance. This review focuses on the mechanisms through which the ncRNAs can be selectively packaged into extracellular vesicles (EVs).

Double Face of eHsp70 in Front of Different Situations

The Hsp70 family is one of the best conserved and abundant member of the heat shock proteins (HSP). This family includes several members and in particular one constitutively expressed member (Hsc70) and another one inducibly expressed under several stress conditions (Hsp70). To date, the intracellular functions of Hsp70 are well defined, and increasing evidences establish its roles in the extracellular environment, such as cytoprotection and immunomodulation. Increasing evidences suggest that several cell types are able to release Hsp70 in the extracellular environment, both under physiological and stress conditions. At the same time many release mechanisms have been identified. This chapte…