RESUMEN
Breast cancer is the most common cancers among women and is one of the main causes of morbidity and mortality in this population. In this study, we aimed to conjugate doxorubicin (DOX), a drug widely used in cancer chemotherapy, and folic acid (FA), a ligand targeted for cancer therapy, to lipid-core nanocapsules (LNC), and evaluate the efficacy of the nanoformulation against triple-negative breast cancer (TNBC) MDA-MB-231 cells that overexpress folate receptors (FRs). We performed cell viability assays, quantitative real-time PCR (qRT-PCR), cell migration assay, and clonogenic assay, as well as measured the levels of nitric oxide (NO) generated and cellular uptake. The results showed that the nanoformulation reduced cell viability. The results of qRT-PCR analysis revealed that the nanoformulation induced apoptosis of MDA-MB-231 cells. The mRNA expression levels of Cat and MnSod were increased when the nanoformulation was compared to the doxorubicin solution. Furthermore, the nanoformulation significantly decreased the migration of breast cancer cells in vitro and inhibited colony formation. Additionally, the expression of iNOS in MDA-MB-231 cells was higher when the nanoformulation was used compared to the doxorubicin solution. Cellular uptake was observed after incubating the MDA-MB-231 cells with the fluorescent-labeled nanoformulation. In conclusion, we developed a promising nanoformulation for the treatment of TNBC. Further studies are necessary to demonstrate the in vivo efficacy of this formulation.
Asunto(s)
Nanocápsulas , Neoplasias de la Mama Triple Negativas , Apoptosis , Línea Celular Tumoral , Doxorrubicina/uso terapéutico , Ácido Fólico , Humanos , Nanocápsulas/uso terapéutico , Neoplasias de la Mama Triple Negativas/tratamiento farmacológicoRESUMEN
PURPOSE: Folic acid-doxorubicin-double-functionalized-lipid-core nanocapsules (LNC-CS-L-Zn+2-DOX-FA) were prepared, characterized, and evaluated in vitro against ovarian and bladder cancer cell lines (OVCAR-3 and T24). METHODS: LNC-CS-L-Zn+2-DOX-FA was prepared by self-assembly and interfacial reactions, and characterized using liquid chromatography, particle sizing, transmission electron microscopy, and infrared spectroscopy. Cell viability and cellular uptake were studied using MTT assay and confocal microscopy. RESULTS: The presence of lecithin allows the formation of nanocapsules with a lower tendency of agglomeration, narrower size distributions, and smaller diameters due to an increase in hydrogen bonds at the surface. LNC-L-CS-Zn+2-DOX-FA, containing 98.00 ± 2.34 µg mL-1 of DOX and 105.00 ± 2.05 µg mL-1 of FA, had a mean diameter of 123 ± 4 nm and zeta potential of +12.0 ± 1.3 mV. After treatment with LNC-L-CS-Zn+2-DOX-FA (15 µmol L-1 of DOX), T24 cells had inhibition rates above 80% (24 h) and 90% (48 h), whereas OVCAR-3 cells showed inhibition rates of 68% (24 h) and 93% (48 h), showing higher cytotoxicity than DOX.HCl. The fluorescent-labeled formulation showed a higher capacity of internalization in OVCAR-3 compared to T24 cancer cells. CONCLUSION: Lecithin favored the increase of hydrogen bonds at the surface, leading to a lower tendency of agglomeration for nanocapsules. LNC-CS-L-Zn+2-DOX-FA is a promising therapeutic agent against tumor-overexpressing folate receptors.
Asunto(s)
Doxorrubicina/administración & dosificación , Portadores de Fármacos/química , Nanocápsulas/química , Neoplasias Ováricas/tratamiento farmacológico , Neoplasias de la Vejiga Urinaria/tratamiento farmacológico , Apoptosis/efectos de los fármacos , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Doxorrubicina/farmacocinética , Composición de Medicamentos/métodos , Liberación de Fármacos , Ensayos de Selección de Medicamentos Antitumorales , Femenino , Ácido Fólico/química , Humanos , Lecitinas/química , Neoplasias Ováricas/patología , Tamaño de la Partícula , Neoplasias de la Vejiga Urinaria/patologíaRESUMEN
Photodynamic therapy (PDT) is a current and innovative technique that can be applied in different areas, such as medical, biotechnological, veterinary, among others, both for the treatment of different pathologies, as well as for diagnosis. It is based on the action of light to activate photosensitizers that will perform their activity on target tissues, presenting high sensitivity and less adverse effects. Therefore, knowing that biotechnology aims to use processes to develop products aimed at improving the quality of life of human and the environment, and optimizing therapeutic actions, researchers have been used PDT as a tool of choice. This review aims to identify the impacts and perspectives and challenges of PDT in different areas of biotechnology, such as health and agriculture and oncology. Our search demonstrated that PDT has an important impact around oncology, minimizing the adverse effects and resistance to chemotherapeutic to the current treatments available for cancer. Veterinary medicine is another area with continuous interest in this therapy, since studies have shown promising results for the treatment of different animal pathologies such as Bovine mastitis, Malassezia, cutaneous hemangiosarcoma, among others. In agriculture, PDT has been used, for example, to remove traces of antibiotics of milk. The challenges, in general, of PDT in the field of biotechnology are mainly the development of effective and non-toxic or less toxic photosensitizers for humans, animals and plants. We believe that there is a current and future potential for PDT in different fields of biotechnology due to the existing demand.
Asunto(s)
Fotoquimioterapia/métodos , Fármacos Fotosensibilizantes/química , Animales , Antibacterianos/química , Antineoplásicos/química , Biotecnología , Bovinos , Inocuidad de los Alimentos , Humanos , Mastitis Bovina/tratamiento farmacológico , Leche , Fármacos Fotosensibilizantes/farmacología , Plantas , Calidad de Vida , Neoplasias Cutáneas/tratamiento farmacológico , Drogas Veterinarias/químicaRESUMEN
Photodynamic therapy (PDT) is an expanding treatment modality due to its minimally invasive localized activity and few adverse effects. This therapy requires photosensitive compounds, which have high sensitivity to light exposure. Thus, in this work, the in vitro antitumor activity of meso-tetra(3- and 4-pyridyl)porphyrins (3-TPyP and 4-TPyP) in metastatic melanoma cell (WM1366 line) and non-tumoral Ovarian lineage Chinese Hamister (CHO) was evaluated using photodynamic process. Cell viability tests, molecular docking, annexin V, confocal microscopy and qRT-PCR were performed. Our results show that both porphyrins inhibited the viability of metastatic melanoma cells when exposed to light and did not alter viability in the dark. In addition, they did not demonstrate cytotoxicity in non-tumor cells. Molecular coupling demonstrated platinum porphyrin affinity for the N-terminal region of APO B-100, LDL receptor, and therefore of the cells under study. Genes such as Caspase 3 and 9, P21, Bax / BCL2, MnSod and GSH showed increased expression. For meta isomer 3-PtTPyP treatment, caspase-9 and caspase-3 expression levels showed a 4.89 and 3.23-fold increase, respectively, while for the para isomer 4-PtTPyP, this change was 3.77 and 12.16-fold, respectively. We also observed an upregulated expression of p21, a protein well-known by its action in cell cycle arrest in a p53-dependent manner. Conclusion: 3-PtTPyP and 4-PtTPyP demonstrated antitumor effect on WM1366 cells, inducing apoptosis and significant alteration of cell cytoskeleton actin. Our work shows that platinum(II) porphyrins may be promising photosensitizers for the treatment of metastatic melanoma by PDT.
Asunto(s)
Portadores de Fármacos/química , Fármacos Fotosensibilizantes/química , Platino (Metal)/química , Porfirinas/química , Animales , Apoptosis/efectos de los fármacos , Sitios de Unión , Células CHO , Caspasa 3/genética , Caspasa 3/metabolismo , Cationes/química , Puntos de Control del Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Cricetinae , Cricetulus , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Luz , Melanoma/metabolismo , Melanoma/patología , Simulación del Acoplamiento Molecular , Fármacos Fotosensibilizantes/farmacología , Proteínas Proto-Oncogénicas c-bcl-2/genética , Proteínas Proto-Oncogénicas c-bcl-2/metabolismoRESUMEN
The global incidence of cancer is rising rapidly and continues to be one of the leading causes of death in the world. Melanoma deserves special attention since it represents one of the fastest growing types of cancer, with advanced metastatic forms presenting high mortality rates due to the development of drug resistance. The aim of this review is to evaluate how the screening of drugs and compounds for melanoma has been performed over the last seven decades. Thus, we performed literature searches to identify melanoma drug screening methods commonly used by research groups during this timeframe. In vitro and in vivo tests are essential for the development of new drugs; however, incorporation of in silico analyses increases the possibility of finding more suitable candidates for subsequent tests. In silico techniques, such as molecular docking, represent an important and necessary first step in the screening process. However, these techniques have not been widely used by research groups to date. Our research has shown that the vast majority of research groups still perform in vitro and in vivo tests, with emphasis on the use of in vitro enzymatic tests on melanoma cell lines such as SKMEL and in vivo tests using the B16 mouse model. We believe that the union of these three approaches (in silico, in vitro, and in vivo) is essential for improving the discovery and development of new molecules with potential antimelanoma action. This workflow would provide greater confidence and safety for preclinical trials, which will translate to more successful clinical trials and improve the translatability of new melanoma treatments into clinical practice while minimizing the unnecessary use of laboratory animals under the principles of the 3R's.