Estrogen receptor positive breast tumors resist chemotherapy by the overexpression of P53 in Cancer Stem Cells.

BACKGROUND AND OBJECTIVES: Breast cancer (BC) is classified according to estrogen receptor (ER) status into ER+ and ER- tumors. ER+ tumors have a worse response to chemotherapy compared to ER- tumors. BCL-2, TP53, BAX and NF-ΚB are involved in drug resistance in the ER+ tumors. Recently it was shown that Cancer Stem Cells (CSCs) play an important role in drug resistance. In this study we tested the hypothesis that CSCs of the ER+ tumors resist drug through the overexpression of BCL-2, TP53, BAX and NF-ΚB. METHODS: CSCs were isolated by anoikis resistance assay from MCF7 (ER+) and MDA-MB-231 (ER-) cell lines. Isolated CSCs were treated with doxorubicin (DOX) and the mRNA expression levels of BCL-2, TP53, BAX and NFKB were investigated by quantitative real time PCR (qPCR) with and without treatment. RESULTS: BCL-2, BAX and NF-ΚB showed decreased expression in MCF7 bulk cancer cells after DOX treatment whereas only BCL-2 and BAX showed decreased expression in MDA-MB-231 bulk cancer cells. Interestingly TP53 was the only gene showed a considerable increase in its expression in CSCs of the ER+ MCF7 cell line compared to bulk cancer cells. Moreover, TP53 was the only gene showing exceptionally higher level of expression in MCF7-CSCs compared to MDA-MB-231-CSCs. CONCLUSION: Our results suggest that CSCs in the ER+ cells escape the effect of DOX treatment by the elevation of p53 expression.

RNAi-mediated knockdown of the voltage gated sodium ion channel TcNav causes mortality in Tribolium castaneum.

The voltage-gated sodium ion channel (VGSC) belongs to the largest superfamily of ion channels. Since VGSCs play key roles in physiological processes they are major targets for effective insecticides. RNA interference (RNAi) is widely used to analyse gene function, but recently, it has shown potential to contribute to novel strategies for selectively controlling agricultural insect pests. The current study evaluates the delivery of dsRNA targeted to the sodium ion channel paralytic A (TcNav) gene in Tribolium castaneum as a viable means of controlling this insect pest. Delivery of TcNav dsRNA caused severe developmental arrest with larval mortalities up to 73% post injection of dsRNA. Injected larvae showed significant (p < 0.05) knockdown in gene expression between 30-60%. Expression was also significantly (p < 0.05) reduced in pupae following injection causing 30% and 42% knockdown for early and late pupal stages, respectively. Oral delivery of dsRNA caused dose-dependant mortalities of between 19 and 51.34%; this was accompanied by significant (p < 0.05) knockdown in gene expression following 3 days of continuous feeding. The majority of larvae injected with, or fed, dsRNA died during the final larval stage prior to pupation. This work provides evidence of a viable RNAi-based strategy for insect control.

High dietborne Cu and Cd induced genotoxicity of Nile tilapia (Oreochromis niloticus).

In this study, the effects of fish diet contaminated with Cu, Cd and Cu+Cd on Nile tilapia, was demonstrated by evaluating its bioaccumulation in the muscle and by testing the cytogenetic profile. Fish exposed to diet contaminated with Cu, Cd or their mixture had a significant increase in the number of chromosomal abnormalities and an inhibition of the mitotic index. Our study revealed high muscle Cu or Cd content in fish fed with diet contaminated with high dietborne Cu, Cd, Cu and Cd. It also revealed that the chromosomal abnormalities were higher for fish fed diet Cd contaminated and Cu+Cd contaminated diets than those fed diet Cu contaminated diet. Thus, maybe fish diets contaminated with Cu, Cd, Cu+Cd induced genotoxicity and mutation. Also, maybe high concentrations of Cu and Cd in fish tissue resulted from feeding on Cu and Cd contaminated diets, are dangerous for human consumption.

Advantages of using mitochondrial 16S rDNA sequences to classify clinical isolates of Acanthamoeba.

PURPOSE: This work was intended to test the classification of Acanthamoeba into genotypes based on nuclear ribosomal RNA gene (18S rDNA, Rns) sequences. Nearly all Acanthamoeba keratitis (AK) isolates are genotype RnsT4. This marked phylogenetic localization is presumably either due to an innate potential for pathogenicity or to a peculiarity of the gene sequences used. To differentiate between these possibilities, relationships among isolates have been reexamined, using a second gene. METHODS: Phylogenetic relationships among isolates of Acanthamoeba were studied, using sequences of the mitochondrial small subunit ribosomal RNA gene (16S rDNA; rns). Genotypes based on complete sequences of approximately 1540 bp were determined for 68 strains, by using multiple phylogenetic analyses. RESULTS: Each strain's mitochondria contained a single intron-free rns sequence (allele). The 68 strains had 35 different sequences. Twenty-eight strains had unique sequences, and 40 strains each shared one of the seven remaining sequences. Eleven mitochondrial rns genotypes corresponding to 11 of 12 previously described nuclear Rns genotypes were identified. Genotype rnsT4 was subdivided into eight distinct clades, with seven including Acanthamoeba keratitis (AK) isolates. CONCLUSIONS: The phylogenetic clustering of AK isolates was confirmed and thus is not specific to the nuclear gene. Rns and rns sequences are both suitable for genotyping of ACANTHAMOEBA: However, the mitochondrial sequences are shorter and more consistent in length, have a higher percentage of alignable bases for sequence comparisons, and have none of the complications caused by multiple alleles or introns, which are occasionally found in Rns. In addition, the more common occurrence of strains with identical rns sequences simplifies identification and clustering of isolates.