The Expression of Markers of Acute Kidney Injury Kim1 and NGAL after Administration of High Doses of Lithium Carbonate in Mice with Engrafted Skin Melanoma B16.

The expression of marker proteins of acute kidney injury after administration of high doses of lithium carbonate was assessed to evaluate the possibility of lithium use in neutron capture therapy. In mice with implanted skin melanoma B16, the expression of Kim1 (kidney injury molecule 1) and NGAL (neutrophil gelatinase-associated lipocalin) proteins in the kidneys was evaluated immunohistochemically 15, 30, 90, 180 min, and 7 days after peroral administration of lithium carbonate at single doses of 300 and 400 mg/kg. An increase in the expression of the studied proteins was found in 30 and 90 min after administration of 400 mg/kg lithium carbonate, however, 7 days after the drug administration, the expression returned to the level observed in the control group. It can be suggested that single administration of lithium carbonate in the studied doses effective for lithium neutron capture therapy will not significantly affect the renal function.

Assessment of Autophagy in Tumor Cells of Human Skin Melanoma of Different Stages.

High levels of autophagy can increase the viability of tumor cells as well as their resistance to chemotherapy. Evaluation of the dynamics of autophagy processes at different stages of carcinogenesis can extend our understanding of melanoma pathogenesis to develop new therapeutic approaches. We performed a comparative study of tumor cell autophagy in stages II and III human skin melanoma. Tumor cells were characterized by high content of structures associated with autophagy (autophagosomes and autolysosomes). In stage III melanoma characterized by the presence of regional metastases in the lymph nodes, tumor cells showed higher expression of the autophagy marker protein LC3beta in comparison with stage II melanoma cells, which can indicate the involvement of autophagy processes in tumor progression and the formation of metastases in the lymph nodes.

Morphological Analysis of Blood Capillaries and Transport Function of Endothelial Cells in Hepatocellular Carcinoma-29.

Hepatocellular carcinoma is characterized by the pronounced vascularization, high levels of vascular endothelial growth factors, and intensive angiogenesis. Caveola-mediated endocytosis and clathrin-dependent endocytosis contribute to internalization of various growth factors, thus affecting blood supply to the tumor and neoangiogenesis. We performed a quantitative analysis of blood vessels in the tumor and evaluated the basal levels of endocytosis-associated vesicular structures in the endothelial cells of the hepatocellular carcinoma-29. The numbers of blood vessels and clathrin-coated vesicles in endothelial cells in the tumor tissue were significantly higher than in normal tissue. The association between clathrin-dependent endocytosis in the endotheliocytes of the capillaries and tumor angiogenesis can provide the basis for the development of new molecular targets for antiangiogenic antitumor therapy.

Ultrastructural Organization of Uveal Melanoma Stromal Cells.

Typical blood capillaries and vessels in uveal melanoma were shown and different types of uveal melanoma stromal cells were determined by electron microscopy and immunohistochemical analysis. Macrophages, fibroblasts of varying degrees of differentiation and endothelial-like cells with numerous caveolae in the cytoplasm were found in the channels of the extracellular matrix surrounding accumulations of tumor cells. The presence local structures positively stained for markers of the blood and lymphatic vessels (CD31 and podoplanin) in channels of the extracellular matrix suggests that the described endothelial-like cells can be the structural basis for blood and lymphatic vessels of the tumor.

Cytological Characteristics of a Heterogeneous Population of Hepatocellular Carcinoma-29 Cells after Injection of Lithium Carbonate in the Experiment.

Five cytological types of hepatocellular carcinoma-29 (G-29) grown in the muscle tissue of the thigh of experimental animals were identified by transmission electron microscopy; 89% of these were poorly differentiated type I-III cells. Lithium in a concentration of 20 mM produced a damaging effect on poorly differentiated G-29 cells: the number of cells with zones of intracellular component destruction and volume density of these zones increased, while volume density of cisterns of endoplasmic reticulum decreased. These results suggest that lithium carbonate can cause destructive changes in the heterogeneous population of G-29 cells during in vivo tumor development.

Ultrastructural Changes in Hepatocellular Carcinoma-29 Cells after Treatment with Lithium Carbonate.

We studied the effect of lithium carbonate on hepatocellular carcinoma-29 cells in CBA male mice after injection in a dose of 20 mM along the tumor periphery. Transmission electron microscopy revealed a decrease in the volume density of the granular endoplasmic reticulum in the cell cytoplasm and an increase in the total numerical and volume density of autophagosomes and autolysosomes and zones of destruction of intracellular organelles. The ability of lithium carbonate to activate intracellular degradation processes in tumor cells and to stimulate cell death can be used to develop new combined strategies in the chemotherapy for hepatocellular carcinoma.

Autophagy in Hepatocytes during Distant Tumor Growth.

Structural changes in the liver of CBA mice were studied during the development of experimental hepatocarcinoma-29 inoculated into the hip. A decrease in the volume density of hepatocyte cytoplasm, mitochondria, endoplasmic reticulum, and lipid inclusions and an increase in the volume density of lysosomal structures during tumor growth were observed. All stages of intracellular autophagy were recorded by the method of electron microscopy. These stages included the appearance of autophagosomes, autophagolysosomes, and secondary lysosomes in the hepatocyte cytoplasm. Fragments of cytoplasm, glycogen rosettes, mitochondria, and fragments of endoplasmic reticulum with ribosomes were found in autophagosomes. The obtained data indicate the development of non-selective autophagy in the liver during distant tumor growth in aimed at the maintenance of intracellular homeostasis in hepatocytes and energy and trophic homeostasis of organism.