cholecalciferol and thiazolyl-blue

Metformin is usually used for the treatment of type 2 diabetes. Recently, many studies suggest that metformin and vitamin D have broad-spectrum antitumor activities. Our aim in this research was to study the effects of vitamin D3 combined with metformin on the apoptosis induction and its mechanisms in the human breast cancer cell line MDA-MB-231. Cell proliferation was measured by methylthiazol tetrazolium (MTT) assay. The morphology of cell apoptosis was observed after Hoechst 33342 staining. Here we show that vitamin D3 280 μg/ml or vitamin D3 300 μg/ml or vitamin D3 320 μg/ml seperately combined with metformin 15000 μg/ml exhibited synergistic effects on cell proliferation and apoptosis. The underlying anti-tumor mechanisms may involve m-TOR related pathways, which are related to activating expression of cleaved caspase-3, Bax and p-AMPK, as well as inhibiting expressions of p-Bcl-2, c-Myc, p-IGF-IR, p-mTOR, p-P70S6K, p-S6.

Topics: Apoptosis; Breast Neoplasms; Cell Line, Tumor; Cell Proliferation; Cholecalciferol; Drug Synergism; Female; Humans; Hypoglycemic Agents; Metformin; Signal Transduction; Tetrazolium Salts; Thiazoles; TOR Serine-Threonine Kinases; Vitamins

To study the effects of vitamin D3 combined with metformin on the proliferation and apoptosis in human bladder cancer cell line SW-780 and its possible mechanism.. MTT assay and fluorescence microscope observations were used to study the effects of vitamin D3 combined with metformin on the proliferation and apoptosis of SW-780 cells in vitro. Western blot was used to detect the expression of apoptosis-related proteins p-Bcl-2, Bax, Cyclin D1, c-Myc and related signaling pathways activated proteins p-IGF-IR, p-mTOR, p-P70S6K, p-S6.. MTT results showed that 320 μg/ml vitamin D3 combined with 620 μg/ml metformin acting on cells for 48h had a significant synergistic effect on proliferation. Fluorescence microscope observations showed that compared with negative control group and monotherapy treatment group, the apoptosis features of combination treatment group were obvious and the apoptosis rate increased greatly. Western blot showed that compared with the negative control group and monotherapy treatment group, the expression levels of p-Bcl-2, Cyclin D1 and c-Myc in combination treatment group significantly decreased, whereas the expression level of Bax significantly increased, and the expression levels of p-IGF-IR, p-mTOR, p-P70S6K and p-S6 in combination treatment group significantly decreased.. Vitamin D3 combined with metformin exhibited obvious inhibitory effects on the cell proliferation and apoptosis induction in SW-780 cells. The underlying anti-tumor mechanism might be related to inhibiting the expressions of p-Bcl-2, Cyclin D1, c-Myc, p-IGF-IR, p-mTOR, p-P70S6K, p-S6 and activating the expression of Bax.

Topics: Apoptosis; Cell Line, Tumor; Cell Proliferation; Cholecalciferol; Drug Synergism; Female; Humans; Hypoglycemic Agents; Metformin; Signal Transduction; Tetrazolium Salts; Thiazoles; TOR Serine-Threonine Kinases; Urinary Bladder Neoplasms; Vitamins

Oral squamous cell carcinoma (OSCC) is responsible for about 90% of oral malignancies and its incidence is increasing. Despite various treatment protocols, survival rate of OSCC is low. Chemotherapy that is used for treating this carcinoma in advanced stages is systemic therapy that destroys carcinogenic cells, and controls tumor metastasis. Chemotherapy is very toxic and has limitations, especially for patients in advanced stages. Considering positive effects of retinoid and vitamin D3 derivatives in treating some carcinomas, we decided to evaluate the effect of combination of these drugs on OSCC. In this study the effects of combination of 5-fluorouracil, 13-cis retinoic acid and vitamin D3 on cultured cell of OSCC have been evaluated.. OSCC cells were cultured in culture media and different concentration of 5-fluorouracil, 13-cis retinoic acid and vitamin D3 were added to cultured cell as separately and in combinations. The effect of treatment on cell proliferation and induction of apoptosis were evaluated by MTT and TUNEL assays respectively.. Combination of 5-fluorouracil and 13- cis retinoic acid had the highest inhibitory effect on SCC cell proliferation. Combination of two drugs had more apoptotic effect than each of them separately, and combination of three drugs had more effect than combination of two drugs.. Because combination of drugs had more inhibitory effect on cell proliferation than one of them and combination of three drugs had the most apoptotic effect than one of these drugs separately, these drugs may have synergic effect on OSCC.. Combination of three drugs has more inhibitory effect on cell proliferation and apoptotic effect than one of these drugs.

Topics: Apoptosis; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Proliferation; Cholecalciferol; Drug Combinations; Drug Synergism; Fluorouracil; Humans; In Situ Nick-End Labeling; Isotretinoin; Mouth Neoplasms; Tetrazolium Salts; Thiazoles

Analogs of 1,25-dihydroxyvitamin D3 with a reversed configuration at C-1 or C-24 and E or Z geometry of the double bond at C-22 in the side chain or at C-5 in the triene system were examined for their antiproliferative activity in vitro against a spectrum of various human cancer cell lines. The analogs coded PRI-2201 (calcipotriol), PRI-2202 and PRI-2205, such as calcitriol and tacalcitol (used as a referential agents), revealed antiproliferative activity against human HL-60, HL-60/MX2, MCF-7, T47D, SCC-25 and mouse WEHI-3 cancer cell lines. The toxicity studies in vivo showed that PRI-2202 and PRI-2205 are less toxic than referential agents. Even at total doses of 2.5-5.0 mg/kg distributed during 5 successive days, no changes in body weight were observed. Calcitriol and tacalcitol showed toxicity in the same protocol at 100 times lower doses. Calcipotriol was lethal to all mice after administration of a total dose of 5.0 mg/kg. The analog PRI-2205 appeared to be more active in mouse Levis lung cancer tumor growth inhibition than calcitriol, calcipotriol or PRI-2202. This analog did not reveal calcemic activity at doses which inhibit tumor growth in vivo nor at higher doses.

Topics: Animals; Antineoplastic Agents; Breast Neoplasms; Calcitriol; Calcium; Carcinoma, Lewis Lung; CD11b Antigen; Cell Cycle; Cell Differentiation; Cell Line, Tumor; Cell Proliferation; Cholecalciferol; Coloring Agents; Female; Fibroblasts; HL-60 Cells; Humans; Lipopolysaccharide Receptors; Male; Mice; Mice, Inbred C57BL; Mice, Inbred DBA; Pancreatic Neoplasms; Prostatic Neoplasms; Rhodamines; Stereoisomerism; Tetrazolium Salts; Thiazoles

Tumor-endothelial interaction contributes to local prostate tumor growth and distant metastasis. In this communication, we designed a novel approach to target both cancer cells and their "crosstalk" with surrounding microvascular endothelium in an experimental hormone refractory human prostate cancer model. We evaluated the in vitro and in vivo synergistic and/or additive effects of a combination of conditional oncolytic adenovirus plus an adenoviral-mediated antiangiogenic therapy. In the in vitro study, we demonstrated that human umbilical vein endothelial cells (HUVEC) and human C4-2 androgen-independent (AI) prostate cancer cells, when infected with an antiangiogenic adenoviral (Ad)-Flk1-Fc vector secreting a soluble form of Flk1, showed dramatically inhibited proliferation, migration and tubular formation of HUVEC endothelial cells. C4-2 cells showed maximal growth inhibition when coinfected with Ad-Flk1-Fc and Ad-hOC-E1, a conditional replication-competent Ad vector with viral replication driven by a human osteocalcin (hOC) promoter targeting both prostate cancer epithelial and stromal cells. Using a three-dimensional (3D) coculture model, we found that targeting C4-2 cells with Ad-hOC-E1 markedly decreased tubular formation in HUVEC, as visualized by confocal microscopy. In a subcutaneous C4-2 tumor xenograft model, tumor volume was decreased by 40-60% in animals treated with Ad-Flk1-Fc or Ad-hOC-E1 plus vitamin D3 alone and by 90% in a combined treatment group, compared to untreated animals in an 8-week treatment period. Moreover, three of 10 (30%) pre-established tumors completely regressed when animals received combination therapy. Cotargeting tumor and tumor endothelium could be a promising gene therapy strategy for the treatment of both localized and metastatic human prostate cancer.

Topics: Adenocarcinoma; Adenoviridae; Angiogenesis Inhibitors; Animals; Antineoplastic Combined Chemotherapy Protocols; Cell Line, Tumor; Cell Proliferation; Cholecalciferol; Endothelial Cells; Genetic Therapy; Genetic Vectors; Humans; Immunohistochemistry; Male; Mice; Mice, Nude; Microscopy, Confocal; Neoplasm Metastasis; Neovascularization, Pathologic; Osteocalcin; Promoter Regions, Genetic; Prostatic Neoplasms; Tetrazolium Salts; Thiazoles; Transplantation, Heterologous; Vascular Endothelial Growth Factor Receptor-2

We determined the in vitro biological activities of 1 alpha, 25-dihdroxyvitamin D(3) (1,25-D(3)) and its analogue, 20-epi-22-oxa-24a, 26a, 27a-trihomo-1 alpha, 25 (OH)(2) vitamin D(3) (KH1060) in six human neuroblastoma (NB) cell lines (SH-SY5Y, NB69, SK-N-AS, IMR5, CHP-134, NGP). The ability of these compounds to inhibit cell growth and DNA synthesis was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay and BrdU incorporation, respectively. The induction of cell death was monitored by caspase-3 activity. Their antineoplastic effect was assessed by clonal proliferation in soft agar. KH1060 was more effective than 1,25 D(3) in inhibiting cell growth and DNA synthesis. The IC-(50) (inhibition of 50% cell viability) indicated that KH1060 was about 10-20-fold more potent than 1,25 D(3). This growth inhibition was also accompanied by induction of caspase-3 activity, indicating that these compounds induce cell death in a caspase-dependent fashion. Moreover, KH1060 exerted potent antineoplastic activity by suppressing the clonal proliferation of the six NB cells. For the first time we demonstrate that KH1060 induces the expression of retinoic acid receptor-beta and p21(Cip1) suggesting that these proteins in part mediate the growth inhibitory effects. Taken together, all the six NB cells were more susceptible to growth inhibition by KH1060 than 1,25-D(3), suggesting its possible use in NB to potentiate the action of retinoids, which are in clinical use for this disease.

Topics: Antineoplastic Agents; Blotting, Western; Brain Neoplasms; Bromodeoxyuridine; Calcitriol; Caspase 3; Caspases; Cell Division; Cell Survival; Cholecalciferol; Coloring Agents; Cyclin-Dependent Kinase Inhibitor p21; Cyclins; DNA; Dose-Response Relationship, Drug; Enzyme-Linked Immunosorbent Assay; Humans; Inhibitory Concentration 50; Models, Chemical; Neuroblastoma; Proto-Oncogene Proteins c-myc; Receptors, Retinoic Acid; Reverse Transcriptase Polymerase Chain Reaction; Tetrazolium Salts; Thiazoles; Tumor Cells, Cultured

1 alpha,25-Dihydroxyvitamin D3 (1 alpha,25(OH)2D3) has recently been reported to exert a toxic effect on both rat and human glioma cell lines. However the potential clinical use of 1 alpha,25(OH)2D3 in the treatment of glioma is impaired by its potent hypercalcemic effects. We have therefore investigated the effects on glioma cell growth of several vitamin D3 analogues which have previously been shown to be less calcemic in vivo than 1 alpha,25(OH)2D3. The present study shows that several analogues are able to induce, in vitro, the death of rat glioma cells (C6.9). The compound KH 1060 appears to be the most effective in the induction of cell death, while MC 1288 and CB 1093 are as potent as 1 alpha,25(OH)2D3. EB 1089 was somewhat less effective than 1 alpha,25(OH)2D3 and MC 903, which is currently used in the treatment of psoriasis, has only a weak activity on C6.9 cells. The effective doses used are around 10(-9) M for 1 alpha,25(OH)2D3 and 10(-10) M for KH 1060. Interestingly, the toxic effect exerted by 1 alpha,25(OH)2D3 and its analogues is accompanied by several of the biochemical features of apoptosis, such as DNA fragmentation and induction of the c-myc protooncogene. These findings, together with the fact that the therapies currently available for glioma are only palliative, suggest that 1 alpha,25(OH)2D3 analogues such as KH 1060, EB 1089 or CB 1093, alone or in combination with other therapeutic approaches, could be of potential interest in the treatment of brain glial tumors.

Topics: Animals; Calcitriol; Calcium; Cell Death; Cholecalciferol; DNA Damage; DNA, Neoplasm; Drug Screening Assays, Antitumor; Gene Expression; Genes, myc; Glioma; Homeostasis; Hypercalcemia; Rats; Tetrazolium Salts; Thiazoles; Tumor Cells, Cultured