37 research outputs found
Oncostatic effect of melatonin in head and neck cancer cells: clonogenic assay
Background: The oncostatic effect of melatonin
has been previously described among different
neoplastic types. One of these is head and neck
squamous cell cancer (HNSCC) with a high rate of
mortality and morbidity. Melatonin (aMT) could cause
cell death in this neoplastic cell type. To determine
this, we performed a clonogenic assay with CAL-27
cells treated with melatonin and/or radiation.
Methods: Cells were plated in a 6-well plate, with 800
cells per well. Assays were performed at least twice
and each time in triplicates. Cells were allowed to
grow 15 days to form colonies of at least 50 cells
each one. Cells were treated with melatonin (100,
500, 1000, 1500 and 2000 ”M) alone or in combination
with irradiation (8 Gy). To visualize colonies, cells
were fixed in 100 % ethanol on days 12, 13, 14 and 15
after they were plated and stained with crystal violet
solution. Colonies were scored with Image J Software.
Results: The results clearly show that melatonin
inhibits colony growth of CAL-27 cells in a dosedependent manner in the groups treated with melatonin
alone 1500 ”M or in combination with irradiation.
Conclusion: The results show the capability of aMT
to prevent colony growth and causing cell death on
CAL-27 cancer cells, especially when combined with
radiation. This is consistent with previous studies on
aMT oncostatic effects and suggests that usage of
aMT in vivo should have future clinical application
Role of c-miR-21, c-miR-126, Redox Status, and Inflammatory Conditions as Potential Predictors of Vascular Damage in T2DM Patients
The development of type 2 diabetes mellitus (T2DM) vascular complications (VCs) is
associated with oxidative stress and chronic inflammation and can result in endothelial dysfunctions.
Circulating microRNAs play an important role in epigenetic regulation of the etiology of T2DM. We
studied 30 healthy volunteers, 26 T2DM patients with no complications, and 26 T2DM patients with
VCs, to look for new biomarkers indicating a risk of developing VCs in T2DM patients. Peripheral
blood samples were used to determine redox state, by measuring the endogenous antioxidant
defense system (superoxide dismutase, SOD; catalase, CAT; glutathione reductase, GRd; glutathione
peroxidase, GPx; and glucose-6-phosphate dehydrogenase, G6DP) and markers of oxidative damage
(advanced oxidation protein products, AOPP; lipid peroxidation, LPO). Additionally, inflammatory
marker levels (IL-1, IL-6, IL-18, and TNF- ), c-miR-21, and c-miR-126 expression were analyzed.
T2DM patients showed the highest oxidative damage with increased GSSG/GSH ratios, LPO, and
AOPP levels. In both diabetic groups, we found that diminished SOD activity was accompanied by
increased CAT and decreased GRd and G6PD activities. Diabetic patients presented with increased
relative expression of c-miR-21 and decreased relative expression of c-miR-126. Overall, c-miR-21,
SOD, CAT, and IL-6 had high predictive values for diabetes diagnoses. Finally, our data demonstrated
that IL-6 exhibited predictive value for VC development in the studied population. Moreover, c-miR-
21 and c-miR-126, along with GPx and AOPP levels, should be considered possible markers for VC
development in future studies.University of GranadaEugenio Rodriguez Pascual Foundation ERP-2021CIBERfes (ISCIII, Spain) CB16-10-0023
The Zebrafish, an Outstanding Model for Biomedical Research in the Field of Melatonin and Human Diseases
The zebrafish has become an excellent model for the study of human diseases because it offers many advantages over other vertebrate animal models. The pineal gland, as well as the biological clock and circadian rhythms, are highly conserved in zebrafish, and melatonin is produced in the pineal gland and in most organs and tissues of the body. Zebrafish have several copies of the clock genes and of aanat and asmt genes, the latter involved in melatonin synthesis. As in mammals, melatonin can act through its membrane receptors, as with zebrafish, and through mechanisms that are independent of receptors. Pineal melatonin regulates peripheral clocks and the circadian rhythms of the body, such as the sleep/wake rhythm, among others. Extrapineal melatonin functions include antioxidant activity, inducing the endogenous antioxidants enzymes, scavenging activity, removing free radicals, anti-inflammatory activity through the regulation of the NF-kappa B/NLRP3 inflammasome pathway, and a homeostatic role in mitochondria. In this review, we introduce the utility of zebrafish to analyze the mechanisms of action of melatonin. The data here presented showed that the zebrafish is a useful model to study human diseases and that melatonin exerts beneficial effects on many pathophysiological processes involved in these diseases.Junta de AndaluciaFEDER Regional Development Fund CTS.205
UGR18
PI 2018 P. Excelencia-P18-RT-69
Oral Mucositis: Melatonin Gel an Effective New Treatment
The current treatment for cervico-facial cancer involves radio and/or chemotherapy. Unfortunately, cancer therapies can lead to local and systemic complications such as mucositis, which is the most common dose-dependent complication in the oral cavity and gastrointestinal tract. Mucositis can cause a considerably reduced quality of life in cancer patients already suffering from physical and psychological exhaustion. However, the role of melatonin in the treatment of mucositis has recently been investigated, and offers an effective alternative therapy in the prevention and/or management of radio and/or chemotherapy-induced mucositis. This review focuses on the pathobiology and management of mucositis in order to improve the quality of cancer patientsâ lives.Part of this study was partially funded by the following grants: SAF2009-14037 from the Spanish Ministry of Economy and Competitivity (MINECO), CB/10/00238 from the Carlos III Health Institute, GREIB.PT_2010_04 from the CEIBiotic Program of the University of Granada, Spain, and CTS-101 from the Innovation, Science, and Business Council, Junta de AndalucĂa, Spain
Protective Effects of Melatonin on the Skin: Future Perspectives
When exposed to hostile environments such as radiation, physical injuries, chemicals,
pollution, and microorganisms, the skin requires protective chemical molecules and pathways.
Melatonin, a highly conserved ancient molecule, plays a crucial role in the maintenance of skin.
As human skin has functional melatonin receptors and also acts as a complete system that is capable of
producing and regulating melatonin synthesis, melatonin is a promising candidate for its maintenance
and protection. Below, we review the studies of new metabolic pathways involved in the protective
functions of melatonin in dermal cells. We also discuss the advantages of the topical use of melatonin
for therapeutic purposes and skin protection. In our view, endogenous intracutaneous melatonin
production, together with topically-applied exogenous melatonin and its metabolites, represent two
of the most potent defense systems against external damage to the skin.Part of this review was co-funded by the Spanish Ministry of the Economy and Competitiveness,
the FEDER Regional Development Fund (nos. SAF2013-49019 and SAF2017-85903), the Charles III Institute
(no. CB/10/00238), and the Economy, Innovation, Science and Employment Council of the Junta de AndalucĂa
(CTS-101). JF and LM are FPU (Professional University formation) Fellows of the Spanish Ministry of Education,
Culture and Sport
Analysis of Plasma MicroRNAs as Predictors and Biomarkers of Aging and Frailty in Humans
Although circulating microRNAs (miRNAs) can modulate gene expression and affect immune system response, little is known
about their participation in age-associated frailty syndrome and sarcopenia. The aim of this study was to determine miRNAs as
possible biomarkers of age and frailty and their correlation with oxidative and inflammatory state in human blood. Three
inflammation-related miRNAs (miR-21, miR-146a, and miR-223) and one miRNA related with the control of melatonin
synthesis (miR-483) were analyzed. Twenty-two healthy adults, 34 aged robust, and 40 aged fragile patients were selected for
this study. The expression of plasma miRNAs was assessed by RT-qPCR; plasma cytokines (IL-6, IL-8, IL-10, and TNFα) were
analyzed by commercial kits, and plasma advanced oxidation protein products (AOPP) and lipid oxidation (LPO) were
spectrophotometrically measured. Fragile subjects had higher miR-21 levels than control subjects, whereas miR-223 and miR-
483 levels increased at a similar extend in both aged groups. All cytokines measured increased in aged groups compared with
controls, without differences between robust and fragile subjects. The fragile group had a TNFα/IL-10 ratio significantly higher
than robust and control groups. Aged groups also had higher AOPP and LPO levels than controls. Women presented higher
AOPP and LPO levels and increased expression of miR-483 compared with men. Positive correlations between miR-21 and
AOPP and between miR-483 and IL-8 were detected. The expression of miR-21 and the TNFα/IL-10 ratio were correlated
positively with the presence of frailty, which suggests that these markers can be considered as possible biomarkers for
age-related frailty.This work was partially supported by grants from the
Ministerio de EconomĂa, Industria y Competitividad y por
el Fondo de Desarrollo Regional Feder, Spain nos. RD12/
0043/0005, PI13-00981, and CB16-10-00238 and from the
Universidad de Granada, Spain no. CEI2014-MPBS3
Intratumoral injection of melatonin enhances tumor regression in cell line-derived and patient-derived xenografts of head and neck cancer by increasing mitochondrial oxidative stress
Head and neck squamous cell carcinoma present a high mortality rate. Melatonin has been shown to have
oncostatic effects in different types of cancers. However, inconsistent results have been reported for in vivo
applications. Consequently, an alternative administration route is needed to improve bioavailability and
establish the optimal dosage of melatonin for cancer treatment. On the other hand, the use of patient-derived
tumor models has transformed the field of drug research because they reflect the heterogeneity of patient
tumor tissues. In the present study, we explore mechanisms for increasing melatonin bioavailability in tumors
and investigate its potential as an adjuvant to improve the therapeutic efficacy of cisplatin in the setting of both
xenotransplanted cell lines and primary human HNSCC. We analyzed the effect of two different formulations of
melatonin administered subcutaneously or intratumorally in Cal-27 and SCC-9 xenografts and in patient-derived
xenografts. Melatonin effects on tumor mitochondrial metabolism was also evaluated as well as melatonin actions
on tumor cell migration. In contrast to the results obtained with the subcutaneous melatonin, intratumoral
injection of melatonin drastically inhibited tumor progression in HNSCC-derived xenografts, as well as in patientderived
xenografts. Interestingly, intratumoral injection of melatonin potentiated CDDP effects, decreasing Cal-
27 tumor growth. We demonstrated that melatonin increases ROS production and apoptosis in tumors, targeting
mitochondria. Melatonin also reduces migration capacities and metastasis markers. These results illustrate the great clinical potential of intratumoral melatonin treatment and encourage a future clinical trial in cancer patients
to establish a proper clinical melatonin treatment.European Regional Development Fund (BâCTSâ071-UGR18)ConsejerĂa de EconomĂa, InnovaciĂłn, Ciencia y Empleo, Junta de AndalucĂa (P18-RTâ32222)Ministerio de Ciencia e InnovaciĂłn/AEI: Agencia Estatal de InvestigaciĂłn/10.13039/501100011033UniĂłn Europea âNextGenerationEUâ/ PRTR (SAF2017-85903âP; PID2020-115112RBâI00)University of Granada (Grant âUNETE,â UCEâPP2017-05)Ministerio de EducaciĂłn Cultura y Deporte, Spai
Melatonin Targets Metabolism in Head and Neck Cancer Cells by Regulating Mitochondrial Structure and Function
This study was funded by grants from the Ministerio de Economia, Industria y Competitividad
y por el Fondo de Desarrollo Regional FEDER, Spain nÂș SAF2013-49019, SAF2017-85903-P,
and from the ConsejerĂa de InnovaciĂłn, Ciencia y Empresa, Junta de AndalucĂa (P07- CTS- 03135,
P10- CTS- 5784, and CTS- 101), Spain. J.F. and L.M. have FPU fellowships from the Ministerio de
EducaciĂłn Cultura y Deporte, Spain. C.R.S. was a schorlarship holder from the Plan Propio de
InvestigaciĂłn of the University of Granada.We wish to thank Michael OâShea for proofreading the paper.Metabolic reprogramming, which is characteristic of cancer cells that rapidly adapt to the hypoxic microenvironment and is crucial for tumor growth and metastasis, is recognized as one of the major mechanisms underlying therapeutic resistance. Mitochondria, which are directly involved in metabolic reprogramming, are used to design novel mitochondria-targeted anticancer agents. Despite being targeted by melatonin, the functional role of mitochondria in melatonin's oncostatic activity remains unclear. In this study, we aim to investigate the role of melatonin in mitochondrial metabolism and its functional consequences in head and neck cancer. We analyzed the effects of melatonin on head and neck squamous cell carcinoma (HNSCC) cell lines (Cal-27 and SCC-9), which were treated with 100, 500, and 1500 mu M of melatonin for 1, 3, and 5 days, and found a connection between a change of metabolism following melatonin treatment and its effects on mitochondria. Our results demonstrate that melatonin induces a shift to an aerobic mitochondrial metabolism that is associated with changes in mitochondrial morphology, function, fusion, and fission in HNSCC. We found that melatonin increases oxidative phosphorylation (OXPHOS) and inhibits glycolysis in HNSCC, resulting in increased ROS production, apoptosis, and mitophagy, and decreased cell proliferation. Our findings highlight new molecular pathways involved in melatonin's oncostatic activity, suggesting that it could act as an adjuvant agent in a potential therapy for cancer patients. We also found that high doses of melatonin, such as those used in this study for its cytotoxic impact on HNSCC cells, might lead to additional effects through melatonin receptors.Ministerio de Economia, Industria y Competitividad y por el Fondo de Desarrollo Regional FEDER, Spain
SAF2013-49019
SAF2017-85903-PJunta de Andalucia
P07-CTS-03135
P10-CTS-5784
CTS-101Ministerio de Educacion Cultura y Deporte, SpainPlan Propio de Investigacion of the University of Granad
Melatonin Enhances Cisplatin and Radiation Cytotoxicity in Head and Neck Squamous Cell Carcinoma by Stimulating Mitochondrial ROS Generation, Apoptosis, and Autophagy
Head and neck cancer is the sixth leading cancer by incidence worldwide. Unfortunately, drug resistance and relapse are the
principal limitations of clinical oncology for many patients, and the failure of conventional treatments is an extremely
demoralizing experience. It is therefore crucial to find new therapeutic targets and drugs to enhance the cytotoxic effects of
conventional treatments without potentiating or offsetting the adverse effects. Melatonin has oncostatic effects, although the
mechanisms involved and doses required remain unclear. The purpose of this study is to determine the precise underlying
mitochondrial mechanisms of melatonin, which increase the cytotoxicity of oncological treatments, and also to propose new
melatonin treatments in order to alleviate and reverse radio- and chemoresistant processes. We analyzed the effects of
melatonin on head and neck squamous cell carcinoma (HNSCC) cell lines (Cal-27 and SCC-9), which were treated with 0.1, 0.5,
1, and 1.5mM melatonin combined with 8 Gy irradiation or 10 ÎŒM cisplatin. Clonogenic and MTT assays, as well as autophagy
and apoptosis, involving flow cytometry and western blot, were performed in order to determine the cytotoxic effects of the
treatments. Mitochondrial function was evaluated by measuring mitochondrial respiration, mtDNA content (RT-PCR), and
mitochondrial mass (NAO). ROS production, antioxidant enzyme activity, and GSH/GSSG levels were analyzed using a
fluorometric method. We show that high concentrations of melatonin potentiate the cytotoxic effects of radiotherapy and
CDDP in HNSCC, which are associated with increased mitochondrial function in these cells. In HNSCC, melatonin induces
intracellular ROS, whose accumulation plays an upstream role in mitochondria-mediated apoptosis and autophagy. Our
findings indicate that melatonin, at high concentrations, combined with cisplatin and radiotherapy to improve its effectiveness,
is a potential adjuvant agent.This study was partially supported by grants from the Ministerio
de EconomĂa y Competitividad, Spain, and the FEDER
Regional Development Fund (nos. SAF2013-49019 and
SAF2017-85903), from the Instituto de Salud Carlos III (no.
CB/10/00238), and from the ConsejerĂa de EconomĂa, InnovaciĂłn,
Ciencia y Empleo, Junta de AndalucĂa (CTS-101)