Catechin hydrate suppresses MCF-7 proliferation through TP53/Caspase-mediated apoptosis

Catechin hydrate (CH), a strong antioxidant that scavenges radicals, is a phenolic compound that is extracted from plants and is present in natural food and drinks, such as green tea and red wine. CH possesses anticancer potential. The mechanism of action of many anticancer drugs is based on their ability to induce apoptosis. In this study, I sought to characterize the downstream apoptotic genes targeted by CH in MCF-7 human breast cancer cells. CH effectively kills MCF-7 cells through induction of apoptosis. Apoptosis was confirmed by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and real-time PCR assays. Cells were exposed to 150 μg/ml CH and 300 μg/mL CH for 24 hours, which resulted in 40.7% and 41.16% apoptotic cells, respectively. Moreover, a 48-hour exposure to 150 μg/ml CH and 300 μg/ml CH resulted in 43.73% and 52.95% apoptotic cells, respectively. Interestingly, after 72 hours of exposure to both concentrations of CH, almost 100% of cells lost their integrity. These results were further confirmed by the increased expression of caspase-3,-8, and -9 and TP53 in a time-dependent and dose-dependent manner, as determined by real-time quantitative PCR. In summary, the induction of apoptosis by CH is affected by its ability to increase the expression of pro-apoptotic genes such as caspase-3, -8, and -9 and TP53

Chloroform Extract of Rasagenthi Mezhugu, a Siddha Formulation, as an Evidence-Based Complementary and Alternative Medicine for HPV-Positive Cervical Cancers

Rasagenthi Mezhugu (RGM) is a herbomineral formulation in the Siddha system of traditional medicine and is prescribed in the southern parts of India as a remedy for all kinds of cancers. However, scientific evidence for its therapeutic efficacy in cervical cancer is lacking, and it contains heavy metals. To overcome these limitations, RGM was extracted, and the fractions were tested on HPV-positive cervical cancer cells, ME-180 and SiHa. The extracts, free from the toxic heavy metals, affected the viability of both the cells. The chloroform fraction (cRGM) induced DNA damage and apoptosis. Mitochondria-mediated apoptosis was indicated. Though both the cells responded to the treatment, ME-180 was more responsive. Thus, this study brings up scientific evidence for the efficacy of RGM against the HPV-mediated cervical cancer cells and, if the toxic heavy metals are the limitation in its use, cRGM would be a suitable candidate as evidence-based complementary and alternative medicine for HPV-positive cervical cancers

Induction of apoptosis in HeLa cells by chloroform fraction of seed extracts of Nigella sativa

<p>Abstract</p> <p>Background</p> <p>Cancer remains one of the most dreaded diseases causing an astonishingly high death rate, second only to cardiac arrest. The fact that conventional and newly emerging treatment procedures like chemotherapy, catalytic therapy, photodynamic therapy and radiotherapy have not succeeded in reverting the outcome of the disease to any drastic extent, has made researchers investigate alternative treatment options. The extensive repertoire of traditional medicinal knowledge systems from various parts of the world are being re-investigated for their healing properties. This study progresses in the direction of identifying component(s) from <it>Nigella sativa </it>with anti cancer acitivity. In the present study we investigated the efficacy of Organic extracts of <it>Nigella sativa </it>seed powder for its clonogenic inhibition and induction of apoptosis in HeLa cancer cell.</p> <p>Results</p> <p>Methanolic, n-Hexane and chloroform extracts of <it>Nigella sativa </it>seedz effectively killed HeLa cells. The IC₅₀values of methanolic, n-hexane, and chloroform extracts of <it>Nigella sativa </it>were 2.28 μg/ml, 2.20 μg/ml and 0.41 ng/ml, respectively. All three extracts induced apoptosis in HeLa cells. Apoptosis was confirmed by DNA fragmentation, western blot and terminal transferase-mediated dUTP-digoxigenin-end labeling (TUNEL) assay.</p> <p>Conclusion</p> <p>Western Blot and TUNEL results suggested that <it>Nigella sativa </it>seed extracts regulated the expression of pro- and anti- apoptotic genes, indicating its possible development as a potential therapeutic agent for cervical cancer upon further investigation.</p

Differential Expression Profile and Genetic Variants of MicroRNAs Sequences in Breast Cancer Patients

The technology available for cancer diagnosis and prognosis is not yet satisfactory at the molecular level, and requires further improvements. Micro RNAs (miRNAs) have been recently reported as useful biomarkers in diseases including cancer. We performed a miRNA expression profiling study using peripheral blood from breast cancer patients to detect and identify characteristic patterns. A total of 100 breast cancer patients and 89 healthy patients were recruited for miRNA genotyping and expression profiling. We found that hs-miR-196a2 in premenopausal patients, and hs-miR-499, hs-miR-146a and hs-miR-196a2 in postmenopausal patients, may discriminate breast cancer patients from healthy individuals. In addition, we found a significant association between two microRNA polymorphisms (hs-miR-196a2 and hs-miR-499) and breast cancer risk. However, no significant association between the hs-miR-146a gene and breast cancer risk was found. In summary, the study demonstrates that peripheral blood miRNAs and their expression and genotypic profiles can be developed as biomarkers for early diagnosis and prognosis of breast cancer

Regulatory mechanisms in the stabilization of p53 tumor suppressor gene in zinc depleted hepatoblastoma cells

The influence of zinc status on the expression of p53, the human tumor suppressor gene, as well as other proteins that may be involved in p53 stability were examined in hepatoblastoma cells (HepG2). Cells were cultured in the zinc-depleted and supplemented media. Chelex 100 resin, a divalent ion-chelating resin, was used to deplete zinc from fetal bovine serum FBS. The Dulbecco's Modified Eagle Medium (DMEM) with 10% chelexed FBS, containing 0.2 μmol/L and 0.4 μmol/L zinc added were termed the zinc deficient ZD0.2 and ZD0.4 media, respectively. The other media consisted of the zinc-normal (ZN), zinc-adequate (ZA), and zinc supplement (ZS) groups, contained 4, 16, and 32 μmol/L zinc, respectively. Cells growth was depressed only in ZD0.2 cells to 78% of ZN cells. As compared to ZN cells, cellular zinc levels were reduced 67% and 56% in ZD0.2 and ZD0.4, respectively, but increased 84% and 127% in ZA and ZS cells, respectively. Nuclear p53 levels were almost 100% and 40% higher in ZD0.2 and ZD0.4 cells, respectively, than ZN cells. In contrast, p53 mRNA abundance was increased 40% in ZD0.4 cells and depressed 60% in ZD0.2 cells as compared with ZN cells. No differences in nuclear p53 protein and p53 mRNA levels were observed among ZN, ZA, and ZS cells. Total cellular and nuclear p21 protein, a major downstream p53 target, as well as p21 mRNA levels were markedly reduced in ZD0.2 and ZD0.4 cells, but were not altered in ZA and ZS cells, when compared to ZN cells. Mdm2 protein, which modulates p53 nuclear export and degradation, was more than twofold higher in the nuclear of ZD0.2 and ZD0.4 cells as compared to ZN cells. In contrast, Mdmx, known to bind Mdm2 and interfere with Mdm2-dependent p53 nuclear export, was depressed in the cytoplasm but not in the nucleus of ZD0.2 and ZD0.4 cells as compared with ZN cells. Moreover, c-Abl, capable of binding mdm2 and enhancing its nuclear accumulation in a p53-independent manner, was not alter in the total but lower in the nucleus of ZD0.2 and ZD0.4 cells than in ZN cells. However, the amount of Mdm2 bound to p53 was depressed and that bound to Mdmx and c-Abl were increased in ZD0.2 and ZD0.4 cells. In zinc deficiency, the reduced binding of Mdm2 to p53 may have resulted from the observed enhanced phosphorylation of p53 at serine 15 and 392, and Mdm2 tyrosine phosphorylation. Most importantly, the accumulation of nuclear p53 in ZD0.2 and ZD0.4 may have resulted from the marked reduction in the nuclear p300, a platform for bringing Mdm2, p53, and other factors together, for p53 nuclear export and degradation

Iron Deficiency in Pre-School Children in Riyadh City

: Iron status was assessed in a representative sample of 189 preschool children living in Riyadh using biochemical and dietary measure. Notably, the mean iron intake for all age groups was lower than dietary reference intake (DRI). In addition, 76% of the children consumed les than the DRI at age 2-3y, meanwhile, only 20.7% of children at age 4-6y consumed higher than the DRI. The hemoglobin concentration ranged between 6.2 and 15.2 g/L with an overall sample mean of 11.7±1.6. In addition, nearly fifty percent (49.3) of children had hemoglobin concentration below 12 g/dL. Similarly, the hematocrit level ranged between 18.5 to 43.9% with an overall sample mean of 36.2±4.4%. The mean serum ferritin level was 18.9 µg/dL. Using hemoglobin concentration as indicators for anemia, 22.7% of children had hemoglobin concentration below 11 g/d, meanwhile 30% of children had hematocrit value below 34%. Moreover, 26.7% of children had plasma ferritin level below 10 µg/dL, suggesting depleted iron stores. In conclusion, these data suggest that preschool children are at high risk for iron deficiency and anima. In addition, iron deficiency is the single most important for the prevalence of anemia which could be attributed to sub-optimal iron intake