A characterization of four B16 murine melanoma cell sublines molecular fingerprint and proliferation behavior

Background: One of the most popular and versatile model of murine melanoma is by inoculating B16 cells in the syngeneic C57BL6J mouse strain. A characterization of different B16 modified cell sub-lines will be of real practical interest. For this aim, modern analytical tools like surface enhanced Raman spectroscopy/scattering (SERS) and MTT were employed to characterize both chemical composition and proliferation behavior of the selected cells. Methods: High quality SERS signal was recorded from each of the four types of B16 cell sub-lines: B164A5, B16GMCSF, B16FLT3, B16F10, in order to observe the differences between a parent cell line (B164A5) and other derived B16 cell sub-lines. Cells were incubated with silver nanoparticles of 50–100 nm diameter and the nanoparticles uptake inside the cells cytoplasm was proved by transmission electron microscopy (TEM) investigations. In order to characterize proliferation, growth curves of the four B16 cell lines, using different cell numbers and FCS concentration were obtained employing the MTT proliferation assay. For correlations doubling time were calculated. Results: SERS bands allowed the identification inside the cells of the main bio-molecular components such as: proteins, nucleic acids, and lipids. An "on and off" SERS effect was constantly present, which may be explained in terms of the employed laser power, as well as the possible different orientations of the adsorbed species in the cells in respect to the Ag nanoparticles. MTT results showed that among the four tested cell sub-lines B16 F10 is the most proliferative and B164A5 has the lower growth capacity. Regarding B16FLT3 cells and B16GMCSF cells, they present proliferation ability in between with slight slower potency for B16GMCSF cells. Conclusion: Molecular fingerprint and proliferation behavior of four B16 melanoma cell sub-lines were elucidated by associating SERS investigations with MTT proliferation assay

Betulin Complex in γ-Cyclodextrin Derivatives: Properties and Antineoplasic Activities in In Vitro and In Vivo Tumor Models

Given the present high incidence of melanoma and skin cancer, interest in potential drugs of plant origin has increased significantly. Pentacyclic lupane-type triterpenes are widely distributed in plants, offering numerous pharmacological benefits. Betulin is an important compound in the bark of Betula pendula Roth and has important therapeutic properties, including antitumor activities. Its biological effect is limited by its poor water solubility, which can be improved by cyclodextrin complexation. The best results have been obtained by using a novel cyclodextrin derivative, octakis-[6-deoxy-6-(2-sulfanyl ethanesulfonate)]-γ-CD. The complexes between betulin and the previously mentioned cyclodextrin were analyzed by scanning electron microscopy (SEM) and differential scanning calorimetry (DSC) and pharmacologically evaluated in vitro (MTT and immunocytochemistry tests) and in vivo in C57BL/6J mice. The solubility of betulin is improved by cyclodextrin complexation, which creates a stable complex that improves the in vitro and in vivo properties of the active compound

An Update On Natural Compounds and Their Modern Formulations for the Management of Malignant Melanoma

The chapter includes a brief presentation of the types of skin cancer. The most aggressive type of skin cancer, melanoma, is discussed from the point of view of incidence, molecular, and immunohistochemical mechanism along with the most important biomarkers for identification. Recent studies containing active phytocompounds with chemopreventive activity pointing toward phytochemicals used for melanoma prevention and therapy are reviewed. Modern physicochemical formulations for the enhancement of bioavailability of some active phytochemicals with chemopreventive activity for malignant melanoma are discussed

New Insights Regarding the Potential Health Benefits of Isoflavones

Isoflavones are a class of plant secondary metabolites, with an estrogen‐like structure presenting a plethora of biological activities. The chapter discusses important facts about this class of phytoestrogens, from biosynthesis to the latest research about their health benefits. The following major points discussed are: biosynthesis, regulation, isolation, metabolism and bioavailability, isoflavones in diet and intake, and new insights regarding the therapeutic effect including cancer chemoprevention. The chapter ends with a mini review of own research of the anti‐inflammatory and chemopreventive activity of isoflavonoid genistein alone and incorporated in modern pharmaceutical formulations. The chapter updates the interested researchers in the field with the latest progress regarding potential health benefits of isoflavones

Cardioprotective Effects of Cultivated Black Chokeberries (<em>Aronia</em> spp.): Traditional Uses, Phytochemistry and Therapeutic Effects

Cardiovascular diseases represent the main cause of morbidity and mortality worldwide. Obesity, sedentary life style, diet, smoking and stress are the principal inducers of hypertension, endothelium dysfunction and insulin resistance in the developed countries. The latest in vitro and in vivo studies on different type of extracts obtained from black-fruited Aronia highlight its excellent cardioprotective actions for the prevention and treatment of cardiovascular and metabolic disorders. So, this chapter aims to bring an up-to-date regarding the antioxidant, anti-inflammatory, anti-atherosclerotic, antiplatelet, blood pressure, glucose and lipid reduction properties of black-fruited Aronia, as a possible new therapeutic strategy for the primary and secondary prevention of cardiovascular pathologies

Phytocompounds Targeting Cancer Angiogenesis Using the Chorioallantoic Membrane Assay

Cancer is the second cause of mortality worldwide. Angiogenesis is an important process involved in the growth of primary tumors and metastasis. New approaches for controlling the cancer progression and invasiveness can be addressed by limiting the angiogenesis process. An increasingly large number of natural compounds are evaluated as angiogenesis inhibitors. The chorioallantoic membrane (CAM) assay represents an in vivo attractive experimental model for cancer and angiogenesis studies as prescreening to the murine models. Since the discovery of tumor angiogenesis, the CAM has been intensively used in cancer research. The advantages of this in vivo technique are in terms of low time-consuming, costs, and a lower number of sacrificed animals. Currently, a great number of natural compounds are being investigated for their effectiveness in controlling tumor angiogenesis. Potential reducing of angiogenesis has been investigated by our group for pentacyclic triterpenes, in various formulations, and differences in their mechanism were registered. This chapter aims to give an overview on a number of phytocompounds investigated using in vitro, murine models and the chorioallantoic membrane assay as well as to emphasize the use of CAM assay in the study of natural compounds with potential effects in malignancies

Lupan-Skeleton Pentacyclic Triterpenes with Activity against Skin Cancer: Preclinical Trials Evolution

Skin cancer is an increasingly frequent pathology, with a dangerous high percentage of malignant melanoma. The use of synthetic chemotherapy raises the problem of severe adverse effects and the development of resistance to treatment. Therefore, the use of natural therapies became the focus of numerous research groups due to their high efficacy and lower systemic adverse effects. Among natural products evaluated as therapeutical agents against skin cancer, betulinic acid was emphasized as a highly selective anti-melanoma agent and is currently undergoing phase II clinical trials as topical application. Several other pentacyclic triterpenes exhibit antiproliferative activities. This chapter aims to present the latest main discoveries in the class of pentacyclic triterenes with antitumor effect and the evolution of their preclinical trials. Furthermore, it includes reports on plant sources containing pentacyclic triterpenes, as well as the main possibilities of their water solubilization and cancer cell targeting. A review on recent data regarding mechanisms of action at cellular and molecular levels complements information on the outstanding medicinal potential of these compounds