Genistein promotes cell death of ethanol-stressed HeLa cells through the continuation of apoptosis or secondary necrosis.

RIGHTS : This article is licensed under the BioMed Central licence at http://www.biomedcentral.com/about/license which is similar to the 'Creative Commons Attribution Licence'. In brief you may : copy, distribute, and display the work; make derivative works; or make commercial use of the work - under the following conditions: the original author must be given credit; for any reuse or distribution, it must be made clear to others what the license terms of this work are.BACKGROUND: Apoptosis is a major target and treatment effect of multiple chemotherapeutical agents in cancer. A soybean isoflavone, genistein, is a well-studied chemopreventive agent and has been reported to potentiate the anticancer effect of some chemotherapeutics. However, its mechanistic basis of chemo-enhancement effect remains to be fully elucidated. METHODS: Apoptotic features of low concentration stressed cancer cells were studied by microscopic method, western blot, immunostaining and annexin V/PI assay. Genistein's effects on unstressed cells and recovering cells were investigated using MTT cell viability assay and LDH cytotoxicity assay. Quantitative real-time PCR was employed to analyze the possible gene targets involved in the recovery and genistein's effect. RESULTS: Low-concentration ethanol stressed cancer cells showed apoptotic features and could recover after stress removal. In stressed cells, genistein at sub-toxic dosage promoted the cell death. Quantitative real-time PCR revealed the up-regulation of anti-apoptotic genes MDM2 and XIAP during the recovery process in HeLa cells, and genistein treatment suppressed their expression. The application of genistein, MDM2 inhibitor and XIAP inhibitor to the recovering HeLa cells caused persistent caspase activity and enhanced cell death. Flow cytometry study indicated that genistein treatment could lead to persistent phosphatidylserine (PS) externalization and necrotic events in the recovering HeLa cells. Caspase activity inhibition shifted the major effect of genistein to necrosis. CONCLUSIONS: These results suggested two possible mechanisms through which genistein promoted cell death in stressed cancer cells. Genistein could maintain the existing apoptotic signal to enhance apoptotic cell death. It could also disrupt the recovering process in caspase-independent manner, which lead to necrotic events. These effects may be related to the enhanced antitumor effect of chemotherapeutic drugs when they were combined with genistein

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The Clinicopathological Significance of miR-133a in Colorectal Cancer

This study determined the expression of microRNA-133a (MiR-133a) in colorectal cancer (CRC) and adjacent normal mucosa samples and evaluated its clinicopathological role in CRC. The expression of miR-133a in 125 pairs of tissue samples was analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) and correlated with patient's clinicopathological data by statistical analysis. Endogenous expression levels of several potential target genes were determined by qRT-PCR and correlated using Pearson's method. MiR-133a was downregulated in 83.2% of tumors compared to normal mucosal tissue. Higher miR-133a expression in tumor tissues was associated with development of distant metastasis, advanced Dukes and TNM staging, and poor survival. The unfavorable prognosis of higher miR-133a expression was accompanied by dysregulation of potential miR-133a target genes, LIM and SH3 domain protein 1 (LASP1), Caveolin-1 (CAV1), and Fascin-1 (FSCN1). LASP1 was found to possess a negative correlation (γ=-0.23), whereas CAV1 exhibited a significant positive correlation (γ=0.27), and a stronger correlation was found in patients who developed distant metastases (γ=0.42). In addition, a negative correlation of FSCN1 was only found in nonmetastatic patients. In conclusion, miR-133a was downregulated in CRC tissues, but its higher expression correlated with adverse clinical characteristics and poor prognosis. © 2014 Timothy Ming-Hun Wan et al.Link_to_subscribed_fulltex

Molecular modifications induced by mud-bath therapy in patients with osteoarthritis

Background Mud-bath therapy (MBT) is a non-pharmacological approach commonly used to treat osteoarthritis (OA). Several data indicate that MBT improve patient's symptoms (1), exerting a beneficial effect on pain and joint function, although the biological mechanisms involved in the therapeutic response are poorly defined. Objectives This study aimed to find molecular changes (proteins and mRNA variations) in patients with OA after MBT treatment. Methods The study included 39 patients whit primary diffuse osteoarthritis, assigned to receive a cycle of mud-bath therapy over a period of 2 weeks added to usual pharmacologic treatment. Whole blood and serum were collected before and after standard MBT treatment: for each time points two pools of patients sera were analyzed by the direct antigen-labeling technology (RayBio® Biotin Label-based Antibody Array, RayBiotech) obtaining a broad, panoramic view of protein expression. Using this semi-quantitative technique up to 1000 target proteins was simultaneously detected, making this approach ideally suited for proteomic studies. Again, pooled samples of mRNA were used to investigate genes expression and to perform the transcriptomic analysis using an high-resolution array design that contains >6.0 million distinct probes, covering coding and non-coding transcripts (GeneChip® HTA 2.0, Affymetrix). Results At the end of mud-bath therapy, using first a semi-quantitative approach, we observed in both biological replicates increased levels (>1.5 fold change) of: inhibin beta A subunit (INHBA), activin A receptor type 2B (ACVR2B), angiopoietin-1 (ANGPT1), beta-2-microglobulin (B2M), growth differentiation factor 10 (BMP-3b/GDF10), C-X-C motif chemokine ligand 5 (CXCL5), fibroblast growth factor 2 (FGF2), fibroblast growth factor 12 (FGF12), oxidized low density lipoprotein receptor 1 (OLR1), matrix metallopeptidase 13 (MMP13). We observed that some increased proteins belongs to the same family or pathway: INHBA, ACVR2B and BMP-3b are members of TGF-Beta superfamily; CXCL5 and MMP13 participate into IL-17 signaling pathway; FGF2 and FGF12 are proteins of FGF family. We are currently performing the transcriptome analysis, and the next step will be to correlate detected proteins with mRNA levels considering also post-transcriptional or epigenetic modifications. Finally we will validate these findings with other quantitative techniques (like ELISA and RT-PCR). Conclusions Our first proteomic and broad spectrum analysis suggest the implication of molecular pathways involved in a wide variety of cellular functions such as gene expression modulation, differentiation, angiogenesis, tissue repair, acute and chronic inflammatory response which may explain the beneficial effects of MBT observed in osteoarthritis. This “omic” approach (proteomic and transcriptomic), generated a huge amount of data that is currently under statistical and bioinformatic analysis. References Forestier R, Desfour H, Tessier JM, et al. Spa therapy in the treatment of knee osteoarthritis: a large randomised multicentre trial. Ann Rheum Dis. 2010;69:660–5

Prognostic factors of overall survival by Cox regression analysis.

<p>Values in parentheses are 95% confidence intervals.</p><p>*Log rank test</p

Chemotherapy has no effect on overall survival of colorectal cancer patients with high or low CD26 expression.

<p>Kaplan-Meier cumulative overall survival curves of CRC patients with high (A) or low (B) CD26 expression. P = 0.514 and 0.661 respectively (log rank test).</p

Significant prognostic factors of overall survival by univariate analysis.

<p>*Log rank test</p

Metastasis associated with worse overall and disease free survival of colorectal cancer patients.

<p>A) Kaplan-Meier cumulative overall survival curves of CRC patients with metastasis and no metastasis. P<0.001 (log rank test). B) Kaplan-Meier disease free survival for CRC patients stratified by metastasis and no metastasis. P<0.001 (log rank test).</p

CD26 expression categorized by clinicopathological data of CRC patients.

<p>TNM, tumour node metastasis.</p>a<p>The total number of cases may be less than 143 because of missing information.</p>b<p>Patients with distant metastasis were classified as “Metastasis”.</p><p>*χ² test (or Fisher exact test in 2-sided)</p