seocalcitol and Body-Weight

1alpha,25-dihydroxyvitamin D3 and its analogue, Seocalcitol (EB1089), are able to reverse or slow the process of carcinogenesis in experimental models and cell cultures. The aim of this study was to investigate the effect of administration vitamin D or Seocalcitol to female Sprague-Dawley rats with 1-methyl-1-nitrosourea (MNU)-induced carcinogenesis of mammary glands on binding characteristics and mRNA levels of thyroid hormone receptors (TRs). Chemopreventive administration of vitamin D caused significant reduction of animal body weight. The expression of TRalpha mRNA was significantly higher in liver of animals treated with vitamin D after detection of first tumour. In our experiment, administration of vitamin D or Seocalcitol significantly reduced KA (group MNU+Seo; MNU+D) and increased Bmax (group MNU+Seo) of thyroid receptors in liver when compared to healthy animals. We show that the activity type I 5'-deiodinase was significantly decreased in livers of animals treated with vitamin D. The data from our in vivo experiment has clearly shown, for the first time, that vitamin D but not Seocalcitol i) may affect the body weight of animals, ii) can cause an increase in the expression of TRalpha in rat liver, remaining the functionality of the TRs unaffected, and iii) is responsible for type I iodothyronine 5'-deiodinase activity decrease in rat liver, remaining the expression of the enzyme unaffected.

Topics: Animals; Body Weight; Breast Neoplasms; Calcitriol; Cholecalciferol; DNA; Female; Gene Expression Regulation; Iodide Peroxidase; Liver; Methylnitrosourea; Organ Size; Rats; Rats, Sprague-Dawley; RNA, Messenger; Thyroid Hormone Receptors alpha

The aim of this study is to determine the effects of 1,25(OH)2D3 and its analogues on tumor growth and body weight, changes in plasma ionized calcium, parathyroid hormone-related protein (PTHrP) production, bone resorption, and the distribution of the 1,25(OH)2D3 receptor (VDR) on tumors in nude mice-bearing the canine adenocarcinoma (CAC-8). Thirty-seven nude mice were implanted subcutaneously with CAC-8. Two weeks after implantation, the mice were divided into 5 groups and injected intraperitoneally 3 times/week for 4 weeks with 5 different substrates. Group I (nontumor-bearing mice) were injected with vehicle. Groups II through V were CAC-8-bearing mice injected with the following: Grp. II, vehicle; Grp. III, analog V; Grp. IV, 1,25(OH)2D3; and Grp. V, EB1089. Our results showed that mice body weight (% change) of CAC-8-bearing mice was significantly lower than those of nontumor-bearing mice (p<0.05). CAC-8-bearing mice treated with analog V maintained their body weight better than CAC-8-bearing mice treated with either vehicle, 1,25(OH)2D3, or EB1089. A reduction of tumor growth was observed in CAC-8-bearing mice treated with 1,25(OH)2D3 and its analogues; however, the reduction was not statistically significant compared to the vehicle-treated CAC-8-bearing mice. All CAC-8-bearing mice increased osteoclastic bone resorption and hypercalcemia. Immunohistochemical staining of CAC-8 with VDR antibody demonstrated a positive reaction in nuclei of tumor cells. In conclusion, CAC-8-bearing mice treated with analog V were more active and maintained their body weight better than other CAC-8-bearing groups. Analog V-treated mice also showed no toxic side effects of hypercalcemia despite an increase in plasmaionized calcium comparable to nontumor-bearing mice. Tumor volumes of CAC-8-bearing mice treated with 1,25(OH)2D3 and its analogues were smaller than vehicle-treated CAC-8-bearing mice. This finding suggested an inhibitory effect on tumor cell growth.

Topics: Adenocarcinoma; Animals; Antineoplastic Agents; Body Weight; Bone and Bones; Bone Resorption; Calcitriol; Calcium; Dogs; Immunoenzyme Techniques; Immunohistochemistry; Mice; Mice, Nude; Neoplasm Transplantation; Parathyroid Hormone-Related Protein; Peptide Hormones; Radioimmunoassay; Receptors, Calcitriol

We examined parathyroid hormone-related peptide (PTHrP) production and regulation in both normal human melanocytes and in a human amelanotic melanoma cell line (A375). Northern blot and immunocytochemical analysis demonstrated that both cultured A375 cells and normal human melanocytes express PTHrP, but A375 cells expressed much higher levels of the peptide. PTHrP secretory rate increased at least 10-fold after treatment with 10% fetal bovine serum (100.2 +/- 2.8 pmol/10(6) cells vs. basal <15 pmol/10(6) cells) in proliferating A375 cells but only twofold in confluent cells. Treatment of A375 cells with increasing concentrations of 1, 25-dihydroxyvitamin D(3) [1,25-(OH)(2)D(3)] or its low-calcemic analog EB-1089 revealed that EB-1089 was 10-fold more potent than 1, 25-(OH)(2)D(3) on inhibition of both cell proliferation and PTHrP expression. Furthermore, inoculation of A375 cells into the mammary fat pad of female severe combined immunodeficiency mice resulted in the development of hypercalcemia and elevated concentrations of plasma immunoreactive PTHrP in the absence of detectable skeletal metastases. Our study, therefore, demonstrates a stepwise increase in PTHrP expression when cells progress from normal to malignant phenotype and suggests that EB-1089 should be further evaluated as a therapeutic agent in human melanoma.

Topics: Animals; Antineoplastic Agents; Blotting, Northern; Body Weight; Calcitriol; Calcium; Cell Division; Cell Line; Drug Implants; Female; Fibroblast Growth Factor 2; Humans; Hypercalcemia; Immunohistochemistry; Insulin; Melanocytes; Melanoma, Amelanotic; Mice; Mice, SCID; Neoplasm Transplantation; Parathyroid Hormone-Related Protein; Proteins; RNA, Messenger; Skin Neoplasms

1,25-Dihydroxyvitamin D3 [1,25-(OH)2D3] and its synthetic analog EB1089 induce characteristic morphological features of apoptosis in MCF-7 cells in vitro that coincide with up-regulation of clusterin and cathepsin B, proteins associated with apoptosis in the mammary gland, and with down-regulation of Bcl-2, an antiapoptotic protein. To determine whether vitamin D3 compounds could mediate apoptosis of breast tumors in vivo, we treated nude mice carrying established MCF-7 xenografts with the low calcemic vitamin D3 analog EB1089 via daily injection or sustained release pellets for up to 5 weeks. The volume of tumors from mice treated with 45 pmol/day EB1089 was 4-fold lower than that of tumors from vehicle-treated control mice after 5 weeks. The reduced growth of tumors from EB1089-treated mice was associated with characteristic apoptotic morphology and a marked reduction in the proportion of epithelial cells to stroma. After 5 weeks of treatment with EB1089, MCF-7 tumors exhibited a 6-fold increase in DNA fragmentation (as measured by in situ end labeling) relative to that in control tumors. The enhanced rate of apoptosis in tumors from EB1089-treated mice was coupled to a 2-fold reduction in proliferation (as measured by expression of proliferating cell nuclear antigen) compared with that in tumors from control mice. The antitumor effects of EB1089 were evident at doses that had minimal effects on serum calcium and body weight. EB1089 treatment did not alter the growth of xenografts derived from a vitamin D3-resistant variant of MCF-7 cells (MCF-7(D3Res) cells), which display resistance to EB1089 in vitro, indicating that resistance to EB1089 is maintained in vivo. Tumors derived from both MCF-7 and MCF-7(D3Res) cells underwent apoptotic regression upon estradiol withdrawal, indicating comparable estrogen dependence of tumors with differential sensitivity to vitamin D3 compounds. These are the first studies to demonstrate apoptotic morphology and regression of human breast tumors in response to treatment with a vitamin D3 analog in vivo and support the concept that vitamin D3 compounds can effectively target human breast cancer.

Topics: Animals; Antineoplastic Agents; Apoptosis; Body Weight; Breast Neoplasms; Calcitriol; Calcium; DNA Fragmentation; Female; Humans; Kinetics; Mammary Neoplasms, Experimental; Mice; Mice, Nude; Mitosis; Neoplasm Transplantation; Transplantation, Heterologous; Tumor Cells, Cultured

EB 1089 is a calcitriol analogue with low calcaemic activity, which inhibits parathyroid hormone-related peptide (PTHrP) secretion in rats implanted with the Leydig cell tumour H-500. We have studied its effect on maternal and fetal plasma PTHrP concentrations and on fetal calcium and phosphorus contents in rats. Three groups of six pregnant rats were injected I.P. daily with EB 1089 (0.10, 0.25 and 0.50 microgram kg-1 in groups 1, 2 and 3, respectively, from day 12 to day 20 of gestation). The control group received an equal volume of solvent alone. On day 21 of gestation the animals were anaesthetized with chloral hydrate and blood samples were taken. Fetuses were collected, weighed and ashed for Ca and P measurements. PTHrP concentrations were measured by radioimmunoassay (RIA) in maternal and fetal plasma. Calcium content (mg (g fetal wt)-1; mean +/- S.E.M.) in control fetuses was not different from that measured in fetuses from dams given the lowest dose of EB 1089, but was higher than that in fetuses from rats injected with EB 1089 at 0.25 microgram kg-1 (1.18 +/- 0.20; P < 0.05) or 0.50 microgram kg-1 (1.08 +/- 0.29; P < 0.05). There was no difference in fetal P content between any of the groups of rats. The PTHrP concentration (pg equivalents human PTHrP(1-34) fragment ml-1) in maternal plasma from control rats (1.70 +/- 0.50) was not different from that in maternal plasma of rats given the lowest dose of EB 1089 (1.44 +/- 0.63).(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Body Weight; Calcitriol; Calcium; Dose-Response Relationship, Drug; Female; Fetal Blood; Fetus; Parathyroid Hormone-Related Protein; Phosphates; Pregnancy; Proteins; Rats; Rats, Wistar

We have examined the effects of 1,25 dihydroxyvitamin D3 (1,25[OH]2D3) and a low calcemic analogue EB1089 on parathyroid hormone-related peptide (PTHRP) production and on the development of hypercalcemia in Fischer rats implanted with the Leydig cell tumor H-500. Leydig cell tumors were implanted subcutaneously into male Fischer rats, which received constant infusions intraperitoneally of either 1,25(OH)2D3 (50-200 pmol/24 h), EB1089 (50-400 pmol/24 h), or vehicle for up to 4 wk. A control group of animals received similar infusions without tumor implantation. Plasma calcium, plasma levels of immunoreactive iPTHRP, and tumor PTHRP mRNA levels were determined as well as tumor size, animal body weight, and animal survival time. Non-tumor-bearing animals receiving > 50 pmol/24 h of 1,25(OH)2D3 became hypercalcemic, whereas no significant change in plasma calcium was observed in animals receiving < or = 200 pmol/24 h of EB1089. Tumor-bearing animals receiving vehicle alone or > 50 pmol/24 h of 1,25(OH)2D3 became severely hypercalcemic within 15 d. However, animals treated with low dose 1,25(OH)2D3 and all doses of EB1089 maintained near-normal or normal levels of plasma calcium for up to 4 wk. Additionally, reduced levels of tumor PTHRP mRNA and of plasma iPTHRP were observed compared with controls in both vitamin D- and EB1089-treated rats. Infusion of 50 pmol/24 h of 1,25(OH)2D3 and 200 pmol/24 h of EB1089 significantly reduced tumor volume by the end of experiment. The analogue but not 1,25(OH)2D3 substantially prolonged survival time in tumor-bearing animals with longer survival achieved at the highest dose, 400 pmol/24 h, of EB1089. These studies demonstrate that 1,25(OH)2D3 and a low calcemic vitamin D analogue are potent inhibitors of PTHRP production in vivo. Low calcemic analogues may therefore represent important alternative therapy for malignancy-associated hypercalcemia.

Topics: Animals; Antineoplastic Agents; Body Weight; Calcitriol; Calcium; Hypercalcemia; Leydig Cell Tumor; Male; Parathyroid Hormone-Related Protein; Proteins; Rats; Rats, Inbred F344; Survival Analysis