ABSTRACT
We aimed to determine the cytotoxic and immunomodulatory effects of hydroalcoholic extracts of the roots and aerial parts of Ebenus boissieri (EB) on breast cancer MDA-MB231 cells and the non-cancerous human embryonic kidney cell line, 293T. Cell viability was determined by MTT assay, trypan blue exclusion, and Live/Dead Viability/Cytotoxicity assay. Apoptosis was evaluated by measuring the activity of caspase-2, 3, 6, 8, and 9. Tumor necrosis factor (TNF)-α and interferon (IFN)-g release was assayed by ELISA, and protein expression of caspase-3, TNF-a, and IFN-g was determined by western blot. The results of this study revealed that MDA-MB231 cell viability was reduced in a dose-dependent manner by the aerial and root extract of EB at 72 h with a half-maximal inhibitory concentration (IC50) of 41.1 ± 2.76 and 65 ± 1.09 µg/mL, respectively. In contrast, neither the aerial nor the root extracts of this plant inhibited the proliferation of 293T cells at doses up to 1000 µg/mL. There was a time-dependent increase in caspase activity, especially caspase-3 and caspase-9. The levels of TNF-aand IFN-g significantly increased in MDA-MB231 cells treated with aerial extract. In conclusion, the extracts of EB induced apoptosis in breast cancer cells by altering the levels of caspases, TNF-a, and IFN-g. The components and precise modes of action of EB have not yet been determined. However, potential antitumor and immunomodulatory activity was observed along with selectivity against cancer cells in vitro, suggesting that hydroalcoholic extracts of this plant are worthy of additional study.
Subject(s)
Antineoplastic Agents/pharmacology , Apoptosis/drug effects , Breast Neoplasms/metabolism , Ebenaceae/chemistry , Immunologic Factors/pharmacology , Plant Extracts/pharmacology , Cell Line, Tumor , Cell Survival/drug effects , HEK293 Cells , Humans , Interferon-gamma/metabolism , Tumor Necrosis Factor-alpha/metabolismABSTRACT
BACKGROUND: Vitamin D receptor (VDR) gene is regarded as one of the candidate genes for type 1 diabetes mellitus (T1D) susceptibility and of some genetic factors involved in the development of osteoporosis in this group. STUDY DESIGN: We characterized the VDR gene polymorphism (BsmI, ApaI, TaqI, FokI and Cdx-2 binding site) in a group of Turkish patients with T1D (n=90) and correlated respective VDR genotypes with the bone mass and some parameters of bone turnover. RESULTS: There were no differences in the genotype frequencies of the BsmI, ApaI, TaqI and Cdx-2 polymorphisms in patients and control subjects. We found a significantly higher prevalence of the F allele/the FF genotype in the patients compared to controls (p=0,0031, odds 1.96 (1,27-3,01)). We observed no difference in markers of bone turnover (Serum levels of osteocalcin, PINP and alkaline phosphatase, urinary levels of calcium/ creatinine and N-telopeptid) among different VDR genotypes. No correlation was found between VDR polymorphisms and DEXA measurements of these patients. CONCLUSIONS: Although the FF genotype was found to be a risk factor in a Turkish population, elucidation of this result is necessary in other larger study groups drawn from the same ethnic population.
