The Foods Containing miR-193b May Inhibit the Growth of Breast Cancer Cells

The plant and animal derived foods contain fats, carbohydrates, proteins and also nucleic acids, such as miRNA. miRNAs are well known the major players in cancer progression as tumor suppressors or oncogenes. Among many miRNAs, miR-193b was investigated the role on cell growth in both of triple-negative and estrogen receptor positive breast cancer cells in this study. For this purpose, cell proliferation was assayed using MTS in MDA-MB-231 and MCF-7 cells treated with 25, 50, 75, 100 nM miR-193b mimic or control miRNA for 72 h. Additionally, colony formation was evaluated by crystal violet dye. Our findings revealed that miR-193b mimic caused to inhibit cell viability as a dose dependent manner in both type of breast cancer cells compare to control condition. Whereas 25 nM dose of miR-193b was not enough to kill, 50 nm led to die 44% and 50% of MCF-7 and MDA-MB-231 cells, respectively. miR-193b mimic concentration above 50 nM, especially 100 nM, resulted in some cytotoxicity after 72 h exposure. These MTS data were also supported by colony formation assay. We strongly believed that the foods containing miR-193b might support to prevent the cell growth of breast cancer cells, even more aggressive type as triple negative

Is the Dietary miR-193b a Novel Cell Cycle Arresting Source for Breast Carcinoma?

There are many reasons that make the foods carcinogenic because of containing inorganic and organic molecules, such as nucleic acids, miRNA. Given the tumor suppressor or oncogenic effects of miRNAs, the dietary carcinogenic effect depends on the type of miRNA. The dietary miRNAs having tumor suppressor properties should be consumed to provide cancer protection and prevention. In this study, we aimed to show the regulatory role of miR-193b on key mediators related to cell cycle pathway in breast cancer cells. We evaluated both mRNA and protein expressions of Cyclin D1 and CDK4 in MCF-7, an estrogen receptor (ER) positive breast cancer cell, and MDA-MB-231, a triple-negative breast cancer cell. Based on our data, miR-193 caused to inhibit the cell cycle at G1 checkpoint, since the expressions of Cyclin D1 and CDK4 were markedly down-regulated at both the mRNA and protein levels in MCF-7 and MDA-MB-231 breast cancer cells after treatment with a 50 nM concentration for 72 h. These data suggest that miR-193b is a tumor suppressor in both ER positive and triple-negative breast cancer cells because it leads the cell cycle arrest at G1 checkpoint. The miR-193b rich foods might help to protect and prevent us from breast cancer

Hypogonadism and erectile dysfunction: an overview

Abstract In humans androgen decline is presented as a clinical picture which includes decreased sexual interest, diminished erectile capasity, delayed or absent orgasms and reduced sexual pleasure. Additionally, changes in mood, diminished well being, fatigue, depression and irritability are also associated with androgen insufficiency. The critical role of androgens on the development, growth, and maintanence of the penis has been widely accepted. Although, the exact effect of androgens on erectile physiology still remains undetermined, recent experimental studies have broaden our understanding about the relationship between androgens and erectile function. Preclinical studies showed that androgen deprivation leads to penile tissue atrophy and alterations in the nerve structures of the penis. Furthermore, androgen deprivation caused to accumulation of fat containing cells and decreased protein expression of endothelial and neuronal nitric oxide synthases (eNOS and nNOS), and phosphodiesterase type-5 (PDE-5), which play crucial role in normal erectile physiology. On the light of the recent literature, we aimed to present the direct effect of androgens on the structures, development and maintanence of penile tissue and erectile physiology as well. Furhermore, according to the clinical studies we conclude the aetiology, pathophysiology, prevalance, diagnosis and treatment options of hypogonadism in aging men. (Asian J Androl 2008 Jan; 10: 36-43) Keywords: testosterone; erectile physiology; symptomatic late onset hypogonadism . Review

Down-regulation of 5-HT1B and 5-HT1D receptors inhibits proliferation, clonogenicity and invasion of human pancreatic cancer cells.

Pancreatic ductal adenocarcinoma is characterized by extensive local tumor invasion, metastasis and early systemic dissemination. The vast majority of pancreatic cancer (PaCa) patients already have metastatic complications at the time of diagnosis, and the death rate of this lethal type of cancer has increased over the past decades. Thus, efforts at identifying novel molecularly targeted therapies are priorities. Recent studies have suggested that serotonin (5-HT) contributes to the tumor growth in a variety of cancers including prostate, colon, bladder and liver cancer. However, there is lack of evidence about the impact of 5-HT receptors on promoting pancreatic cancer. Having considered the role of 5-HT-1 receptors, especially 5-HT1B and 5-HT1D subtypes in different types of malignancies, the aim of this study was to investigate the role of 5-HT1B and 5-HT1D receptors in PaCa growth and progression and analyze their potential as cytotoxic targets. We found that knockdown of 5-HT1B and 5-HT1D receptors expression, using specific small interfering RNA (siRNA), induced significant inhibition of proliferation and clonogenicity of PaCa cells. Also, it significantly suppressed PaCa cells invasion and reduced the activity of uPAR/MMP-2 signaling and Integrin/Src/Fak-mediated signaling, as integral tumor cell pathways associated with invasion, migration, adhesion, and proliferation. Moreover, targeting 5-HT1B and 5-HT1D receptors down-regulates zinc finger ZEB1 and Snail proteins, the hallmarks transcription factors regulating epithelial-mesenchymal transition (EMT), concomitantly with up-regulating of claudin-1 and E-Cadherin. In conclusion, our data suggests that 5-HT1B- and 5-HT1D- mediated signaling play an important role in the regulation of the proliferative and invasive phenotype of PaCa. It also highlights the therapeutic potential of targeting of 5-HT1B/1D receptors in the treatment of PaCa, and opens a new avenue for biomarkers identification, and valuable new therapeutic targets for managing pancreatic cancer

sChronic administration of sildenafil improves erectile function in a rat model of chronic renal failure

The relationship between erectile dysfunction (ED) and chronic renal failure (CRF) has been reported in several studies. This study aimed to investigate whether the chronic use of sildenafil could enhance the erectile capacity in CRF-induced rats. In addition, we assessed the effect of that treatment on certain molecules, which have been suggested to play crucial roles in erectile physiology and CRF-related ED as well. Three groups of animals were utilized: (1) age-matched control rats, (2) CRF-induced rats, (3) CRF-induced rats treated with chronic administration of sildenafil (5 mg kg−1 p.o. for 6 weeks [treatment started after 6 weeks of CRF induction]). At 3 months, all animals underwent cavernosal nerve stimulation (CNS) to assess erectile function. Penile tissue advanced glycation end products (AGE′s)/5-hydroxymethyl-2-furaldehyde, malondialdehyde (MDA), cGMP (ELISA), inducible nitric oxide synthase (iNOS) and neuronal NOS (nNOS) (Western blot) analyses were performed in all rat groups. CRF-induced rats had a significant decrease in erectile function when compared to control rats (P < 0.05). The increase in both intracavernosal pressure (ICP) and area under the curve of CRF-induced rats treated with sildenafil (Group 3) was greater than CRF-induced rats (Group 2). Additionally, sildenafil treatment decreased AGE, MDA and iNOS levels, while it preserved nNOS and cGMP contents in CRF-induced penile tissue. Decreased AGE, MDA, iNOS and increased nNOS, cGMP levels at the sildenafil-treated group increased both ICP and Total ICP to CNS, which led to improve erectile function in CRF-induced rats. The results of the present study revealed the therapeutic effect of chronic sildenafil administration on erectile function in CRF-induced rats

Down-Regulation of 5-HT_1B and 5-HT_1D Receptors Inhibits Proliferation, Clonogenicity and Invasion of Human Pancreatic Cancer Cells

<div><p>Pancreatic ductal adenocarcinoma is characterized by extensive local tumor invasion, metastasis and early systemic dissemination. The vast majority of pancreatic cancer (PaCa) patients already have metastatic complications at the time of diagnosis, and the death rate of this lethal type of cancer has increased over the past decades. Thus, efforts at identifying novel molecularly targeted therapies are priorities. Recent studies have suggested that serotonin (5-HT) contributes to the tumor growth in a variety of cancers including prostate, colon, bladder and liver cancer. However, there is lack of evidence about the impact of 5-HT receptors on promoting pancreatic cancer. Having considered the role of 5-HT-1 receptors, especially 5-HT_1B and 5-HT_1D subtypes in different types of malignancies, the aim of this study was to investigate the role of 5-HT_1B and 5-HT_1D receptors in PaCa growth and progression and analyze their potential as cytotoxic targets. We found that knockdown of 5-HT_1B and 5-HT_1D receptors expression, using specific small interfering RNA (siRNA), induced significant inhibition of proliferation and clonogenicity of PaCa cells. Also, it significantly suppressed PaCa cells invasion and reduced the activity of uPAR/MMP-2 signaling and Integrin/Src/Fak-mediated signaling, as integral tumor cell pathways associated with invasion, migration, adhesion, and proliferation. Moreover, targeting 5-HT_1B and 5-HT_1D receptors down-regulates zinc finger ZEB1 and Snail proteins, the hallmarks transcription factors regulating epithelial-mesenchymal transition (EMT), concomitantly with up-regulating of claudin-1 and E-Cadherin. In conclusion, our data suggests that 5-HT_1B– and 5-HT_1D–mediated signaling play an important role in the regulation of the proliferative and invasive phenotype of PaCa. It also highlights the therapeutic potential of targeting of 5-HT_1B/1D receptors in the treatment of PaCa, and opens a new avenue for biomarkers identification, and valuable new therapeutic targets for managing pancreatic cancer.</p></div

The postulated molecular regulation of 5-HT_1B and 5-HT_1D receptors to β1 integrin/Src/FAK complex, ECM/uPAR/MMP-2 signaling and the zinc finger transcriptional regulators of EMT in PaCa cells.

<p>These receptors-regulated signaling might be mediated through TG2/NF-κB axis. 5-HT_1B and 5-HT_1D receptors mediate β1-integrin activity to recruit a Src–FAK complex promoting the cell proliferation and migration. Upon different extracellular mitogenic stimuli (e.g; growth factors, hormones and neurotransmitters), the over-expressed 5-HT_1B/1D receptors promote the activation of urokinase plasminogen activator receptor (uPAR), and matrix metalloproteinase (MMP-2), facilitating extra-cellular matrix degradation and enhancement of invasion process. Also, 5-HT_1B/1D receptors stimulate the expression of zinc finger transcriptional factors (Snail and TCF8/ZEB1). These mesenchyme markers, in turn, repress the gene transcription of epithelial markers (claudin-1 and E-cadherin) leading to stimulation of epithelial mesenchymal transition (EMT), further supporting the proliferation and invasiveness of PaCa cells.</p

Effect of down-regulation of 5-HT_1B and 5-HT_1D receptors expression on the invasion/migration of PaCa cells.

<p>PANC-1 cells (<b>A</b>) and MIAPaCa-2 cells (<b>B</b>) were transfected with 50 nM of indicated siRNAs (for 72 h), and equal numbers of viable cells were seeded onto Matrigel-coated Transwell filters in Matrigel invasion chambers. The number of the cells that invaded after 24 h was determined as in protocol. Magnification,100×. The histograms show the mean of percentages of invasion ± SD of three experiments. * P<0.05 vs. control cells. (<b>C</b>) The involvement of 5-HT_1B and 5-HT_1D receptors in regulation of PANC-1 cell motility as analyzed by the wound healing assay. A single scratch was made in the center of the confluent cell monolayer, and the wounded monolayers were transfected with indicated siRNAs. The wounds repair was monitored for 24 h and visualized microscopically with original magnification ×100. Images were taken immediately (0 h), and after 12 h and 24 h of scratching the cultures. The histogram shows the percentages of the cells migration, and the data is expressed as mean of the percentages of migration ± SD of three independent experiments. * represents significant difference between indicated groups (P<0.05).</p

Effects of down-regulation of 5-HT_1B and 5-HT_1D receptors on PaCa cell proliferation.

<p>(<b>A</b>) 5-HT_1B and 5-HT_1D receptors are highly expressed in several pancreatic cancer cell lines. Cell lysates of several PaCa cell lines and normal human pancreatic duct epithelial (HPDE) cells were subjected to Western blot analysis as indicated. β-actin was used as a loading control. (<b>B–C</b>) 5-HT_1B and 5-HT_1D receptors expression levels in PANC-1 (B) and MIAPaCa-2 cells (C) after knockdown of the receptors expression with their corresponding siRNAs. Cells were transfected with 50 nM of indicated siRNAs, and 72 h later, cell lysates were subjected to Western blot analysis. β-actin was used as loading control. The histograms show the relative quantification of the indicated proteins levels. (<b>D–E</b>) mRNA expression of 5-HT_1B and 5-HT_1D receptors in PANC-1 (D) and MIAPaCa-2 cells (E) after knockdown of the receptors expression with their corresponding siRNAs. Cells were transfected with 50 nM of indicated siRNAs, and 72 h later, the total RNA was extracted and the transcript levels of 5-HT_1B and 5-HT_1D were determined by standard RT-PCR as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105245#s2" target="_blank">Materials and Methods</a>. GAPDH was used as loading control. (<b>F–G</b>) siRNA-mediated 5-HT_1B and 5-HT_1D receptors knockdown inhibits PaCa cells proliferation. PANC-1 (F) and MIAPaCa-2 cells (G) were transfected with 50 nM of indicated siRNAs, and after 72 h, proliferation was detected by an MTS assay. Data are represented as mean ± SD. * P<0.05 vs. control cells. All experiments were independently performed three times.</p

5-HT_1B/1D receptors-induced regulation of TG2/NF-κB signaling might mediate proliferation/invasion-promoting pathways.

<p>(<b>A</b>) 5-HT_1B and 5-HT_1D receptors regulate TG2 and NF-κB expression. PANC-1 cells were treated with indicated siRNAs as described above. (<b>B</b>) siRNA-mediated TG2 knockdown induced downstream molecular effects similar to that observed after knockdown of 5-HT_1B and 5-HT_1D receptors expression. Cells were transfected with 50 nM of TG2 siRNA or control siRNA, and cell lysates were subjected to Western blot analysis. β-actin was used as loading control. (<b>C</b>) Inhibition of constitutive activation of NF-κB inhibits the key signaling promoting proliferation/invasion and decreases the mesenchymal markers, Fibronectin and α-SMA. Cells were treated with indicated concentration of NF-κB activation Inhibitor II, JSH-23, for 24 h, and cell lysates were subjected to Western blot analysis. Tubulin was used as loading control. Three independent experiments were performed with similar results, and representative data is shown.</p