Activation Of Cell Signaling Pathways Research Articles

RNA sequencing and gene co-expression network of in vitro matured oocytes and blastocysts of buffalo.

In reproductive technologies, uncovering the molecular aspects of oocyte and embryo competence under different conditions is crucial for refining protocols and enhancing efficiency. RNA-seq generates high-throughput data and provides transcriptomes that can undergo additional computational analyses. This study presented the transcriptomic profiles of in vitro matured oocytes and blastocysts produced in vitro from buffalo crossbred (Bubalus bubalis), coupled with gene co-expression and module preservation analysis. Cumulus Oophorus Complexes, obtained from slaughterhouse-derived ovaries, were subjected to in vitro maturation to yield metaphase II oocytes (616) or followed in vitro fertilization and culture to yield blastocysts for sequencing (526). Oocyte maturation (72%, ±3.34 sd) and embryo development (21.3%, ±4.18 sd) rates were obtained from three in vitro embryo production routines following standard protocols. Sequencing of 410 metaphase II oocytes and 70 hatched blastocysts (grade 1 and 2) identified a total of 13,976 genes, with 62% being ubiquitously expressed (8,649). Among them, the differentially expressed genes (4,153) and the strongly variable genes with the higher expression (fold-change above 11) were highlighted in oocytes (BMP15, UCHL1, WEE1, NLRPs, KPNA7, ZP2, and ZP4) and blastocysts (APOA1, KRT18, ANXA2, S100A14, SLC34A2, PRSS8 and ANXA2) as representative indicators of molecular quality. Additionally, genes exclusively found in oocytes (224) and blastocysts (2,200) with specific biological functions were identified. Gene co-expression network and module preservation analysis revealed strong preservation of functional modules related to exosome components, steroid metabolism, cell proliferation, and morphogenesis. However, cell cycle and amino acid transport modules exhibited weak preservation, which may reflect differences in embryo development kinetics and the activation of cell signaling pathways between buffalo and bovine. This comprehensive transcriptomic profile serves as a valuable resource for assessing the molecular quality of buffalo oocytes and embryos in future in vitro embryo production assays.

A Multi-Color Immunofluorescence Assay to Distinguish Intracellular From External Leishmania Parasites.

Leishmaniasis, a neglected tropical disease, is caused by the intracellular protozoan parasite Leishmania. Upon its transmission through a sandfly bite, Leishmania binds and enters host phagocytic cells, ultimately resulting in a cutaneous or visceral form of the disease. The limited therapeutics available for leishmaniasis, in combination with this parasite's techniques to evade the host immune system, call for exploring various methods to target this infection. To this end, our laboratory has been characterizing how Leishmania is internalized by phagocytic cells through the activation of multiple host cell signaling pathways. This protocol, which we use routinely for our experiments, delineates how to infect mammalian macrophages with either promastigote or amastigote forms of the Leishmania parasite. Subsequently, the number of intracellular parasites, external parasites, and macrophages can be quantified using immunofluorescence microscopy and semi-automated analysis protocols. Studying the pathways that underlie Leishmania uptake by phagocytes will not only improve our understanding of these host-pathogen interactions but may also provide a foundation for discovering additional treatments for leishmaniasis. Key features • This protocol visualizes and quantifies multiple intracellular forms of Leishmania. • It offers flexibility at various points for researchers to introduce modifications according to their study needs.

PYGO2 increases proliferation and migration capacities through critical signaling pathways in esophageal squamous cell carcinoma.

Esophageal cancer, an increasingly prevalent malignancy, is a major concern for global health. The development of esophageal squamous cell carcinoma (ESCC) involves various genetic abnormalities that affect key cell signaling pathways, including Wnt, Hh, Apoptosis, MAPK, EGFR, AKT, Notch, and EMT. Additionally, this malignancy involves some changes in the expression of long noncoding RNAs (LncRNAs). The present study examines the relationship between PYGO2 gene expression and the activity of cell signaling pathways in KYSE-30 and YM-1ESCC cell lines. To this end, several cellular and molecular tests were performed, including cell migration, cell cycle, and apoptosis. Also, expression levels of CD133 and CD44 markers, real-time PCR, and western blot were analyzed after inducing PYGO2 protein expression in the cells. Overexpression of the PYGO2 protein resulted in the upregulation of Wnt pathway-related genes, leading to enhanced cell migration and proliferation and reduced apoptosis in both cell lines. Furthermore, PYGO2 gene expression induction analysis showed the correlation of several involved genes in Wnt, Hh, Apoptosis, MAPK, EGFR, AKT, and EMT pathways with various LncRNAs.

Oxidative stress and inflammation induced by air pollution-derived PM2.5 persist in the lungs of mice after cessation of their sub-chronic exposure

More than 7 million early deaths/year are attributable to air pollution. Current health concerns are especially focused on air pollution-derived particulate matter (PM). Although oxidative stress-induced airway inflammation is one of the main adverse outcome pathways triggered by air pollution-derived PM, the persistence of both these underlying mechanisms, even after exposure cessation, remained poorly studied. In this study, A/JOlaHsd mice were also exposed acutely (24 h) or sub-chronically (4 weeks), with or without a recovery period (12 weeks), to two urban PM2.5 samples collected during contrasting seasons (i.e., autumn/winter, AW or spring/summer, SS). The distinct intrinsic oxidative potentials (OPs) of AW and SS PM2.5, as evaluated in acellular conditions, were closely related to their respective physicochemical characteristics and their respective ability to really generate ROS over-production in the mouse lungs. Despite the early activation of the nuclear factor erythroid 2-related factor 2 (Nrf2) cell signaling pathway by AW and, in a lesser degree, SS PM2.5, in the murine lungs after acute and sub-chronic exposures, the critical redox homeostasis was not restored, even after the exposure cessation. Accordingly, an inflammatory response was reported through the activation of the nuclear factor-kappa B (NF-κB) cell signaling pathway activation, the secretion of cytokines, and the recruitment of inflammatory cells, in the murine lungs after the acute and sub-chronic exposures to AW and, in a lesser extent, to SS PM2.5, which persisted after the recovery period. Taken together, these original results provided, for the first time, new relevant insights that air pollution-derived PM2.5, with relatively high intrinsic OPs, induced oxidative stress and inflammation, which persisted admittedly at a lower level in the lungs after the exposure cessation, thereby contributing to the occurrence of molecular and cellular adverse events leading to the development and/or exacerbation of future chronic inflammatory lung diseases and even cancers.

Intermittent fasting against cancer development and progression: Highlighting potential anticancer molecular mechanisms.

Intermittent fasting (IF) diets have been popular since the last few decades because of their provable clinical efficiency on weight control of the subjects. These diet types are generally safe, resulting in health promoting effects against several human diseases like cardiovascular diseases, diabetes mellitus, neurogenerative disorders and cancer. To review whether IF can act against cancer development and progression, highlighting potential anticancer molecular mechanisms in clinical studies. Applied summarization of the available clinical studies investigating the effectiveness of IF against cancer development and progression and cancer-induced indicators. Scientific databases, e.g., PubMed, and Scopus, were comprehensively searched using relative words to identify in vivo and in vitro data, as well as clinical studies. IF seems to exert health-promoting effects in cancer patients through induction of autophagy, which enhances the in vivo suppression of tumor development, by chemotherapy. IF provokes tumors to chemotherapy and defends the normal cells from its adverse side effects, increasing the immune response. In addition, it enhances the cytotoxic CD8(+) tumor-infiltrating lymphocytes and the bone marrow lymphoid progenitor cells, delaying the cancer progression. IF reduces oxidative stress via repression of translation and induces cellular apoptosis. Fasting exerts anti-aging properties modulating the secretion of IGF-1, IGFBP-1, glucose, and insulin while, at the same time, it integrates cell adaptive responses and activates cell signaling pathways which stimulates antioxidant defenses, DNA repairment, control of protein quality, mitochondrial synthesis while decreasing inflammation. IF appears to exert health promoting effects against cancer development and progression, suppressing several kinds of cancer. There are well-recognized and not well-recognized molecular processes accentuating its anticancer outcomes; however, well-designed clinical trials and further molecular studies are strongly recommended.

Two synthetic steroid analogs reduce human respiratory syncytial virus replication and the immune response to infection both in vitro and in vivo

HRSV is responsible for many acute lower airway infections and hospitalizations in infants, the elderly and those with weakened immune systems around the world. The strong inflammatory response that mediates viral clearance contributes to pathogenesis, and is positively correlated with disease severity. There is no specific effective therapy on hand. Antiviral synthetic stigmastanes (22S, 23S)-22,23-dihydroxystigmast-4-en-3-one (Compound 1) and 22,23-dihydroxystigmasta-1,4-dien-3-one (Compound 2) have shown to be active inhibiting unrelated virus like Herpes Simplex type 1 virus (HSV-1) and Adenovirus, without cytotoxicity. We have also shown that Compound 1 modulates the activation of cell signaling pathways and cytokine secretion in infected epithelial cells as well as in inflammatory cells activated by nonviral stimuli. In the present work, we investigated the inhibitory effect of both compounds on HRSV replication and their modulatory effect on infected epithelial and inflammatory cells. We show that compounds 1 and 2 inhibit in vitro HRSV replication and propagation and reduce cytokine secretion triggered by HRSV infection in epithelial and inflammatory cells. The compounds reduce viral loads and inflammatory infiltration in the lungs of mice infected with HRSV.

Clinical, Pathologic, and Molecular Characterization of Acanthosis Nigricans Maligna: A Comprehensive Analysis of a Dermatologic Entity and its Association with Underlying Neoplastic Conditions

Acanthosis nigricans maligna (ANM) is a distinctive dermatologic entity presenting with cutaneous hyperpigmentation and epidermal thickening, characteristically located in flexural areas and skin folds. Despite its apparent cutaneous nature, ANM is frequently associated with underlying neoplastic conditions, implying profound clinical significance. In this study, we undertook a comprehensive evaluation of ANM from clinical, pathological and molecular perspectives with the aim of elucidating its etiopathogenic mechanisms and its relationship with neoplastic processes. From a clinical standpoint, we retrospectively analyzed a cohort of patients diagnosed with ANM, exploring the clinical and epidemiologic features that could indicate the presence of an underlying malignant condition. In addition, histopathologic studies were performed on samples of affected skin to discern the microscopic alterations associated with ANM and their correlates with neoplastic processes. In a molecular phase, an in-depth analysis of genetic and molecular markers in skin samples and associated tumor tissues was carried out using advanced sequencing techniques. This allowed the identification of specific mutations, chromosomal alterations and gene expression profiles that could contribute to both the occurrence of ANM and the promotion of the microenvironment conducive to tumor development. The results obtained indicate a significant correlation between the presence of ANM and the abnormal activation of cell signaling pathways, as well as the altered expression of regulatory factors of cell growth and differentiation. Our study highlights the importance of considering ANM as a potential cutaneous marker of underlying neoplastic processes and suggests that its early detection could facilitate the early identification of early stage malignant conditions. Taken together, this comprehensive analysis provides an in-depth view of ANM and its clinical, pathological and molecular relevance in the context of neoplastic diseases.

An updated review on cell signaling pathways regulated by candidate miRNAs in coronary artery disease

MicroRNAs (miRNAs) are small endogenous non-coding RNA, size range from 17 to 25 nucleotides that regulate gene expression at the post-transcriptional level. More than 2000 different types of miRNAs have been identified in humans which regulate about 60% of gene expression, since the discovery of the first miRNA in 1993. MicroRNA performs many functions such as being involved in the regulation of various biological pathways for example cell migration, cell proliferation, cell differentiation, disease progression, and initiation. miRNAs also play an important role in the development of atherosclerosis lesions, cardiac fibroblast, cardiac hypertrophy, cancer, and neurological disorders. Abnormal activation of many cell signaling pathways has been observed in the development of coronary artery disease. Abnormal expression of these candidate miRNA genes leads to up or downregulation of specific genes, these specific genes play an important role in the regulation of cell signaling pathways involved in coronary artery disease. Many studies have found that miRNAs play a key role in the regulation of crucial signaling pathways that are involved in the pathophysiology of coronary artery disease. This review is designed to investigate the role of cell signaling pathways regulated by candidate miRNAs in Coronary artery disease.

Spatially activated conserved auxin-transcription factor regulatory module controls de novo root organogenesis in rice.

This study reveals that the process of crown root development and auxin-induced de novo root organogenesis during in vitro plantlet regeneration share a common auxin-OsWOX10 regulatory module in rice. In the fibrous-type root system of rice, the crown roots (CR) are developed naturally from the shoot tissues. Generation of robust auxin response, followed by activation of downstream cell fate determinants and signaling pathways at the onset of crown root primordia (CRP) establishment is essential for new root initiation. During rice tissue culture, embryonic calli are induced to regenerate shoots in vitro which undergo de novo root organogenesis on an exogenous auxin-supplemented medium, but the mechanism underlying spatially restricted root organogenesis remains unknown. Here, we reveal the dynamics of progressive activation of genes involved in auxin homeostasis and signaling during initiation and outgrowth of rice crown root primordia. By comparative global dataset analysis, we identify the crown root primordia-expressed genes whose expression is also regulated by auxin signaling. In-depth spatio-temporal expression pattern analysis shows that the exogenous application of auxin induces a set of key transcription factors exclusively in the spatially positioned CRP. Further, functional analysis of rice WUSCHEL-RELATED HOMEOBOX 10 (OsWOX10) during in vitro plantlet regeneration from embryogenic calli shows that it promotes de novo root organogenesis from regenerated shoots. Expression of rice OsWOX10 also induces adventitious roots (AR) in Arabidopsis, independent of homologous endogenous Arabidopsis genes. Together, our findings reveal that a common auxin-transcription factor regulatory module is involved in root organogenesis under different conditions.

Enhancements of Bcl-2/mTOR/ERK1/2 activities by antioxidant mechanisms confer cardioprotection on Ginkgo biloba supplement against isoprenaline-induced myocardial infarction in rats

BackgroundMyocyte necrosis and apoptosis can be severely induced by acute adrenergic stimulation due to the activation of several cell signalling pathways. However, uncertainty exists regarding the potential cardioprotective effects of Ginkgo biloba (GBS), a Chinese herbal supplement rich with antioxidants, on isoprenaline (ISO)-induced myocardial infarction. Therefore, this work was designed to investigate the cardioprotective mechanisms of GBS on ISO-induced myocardial infarction. MethodsAnimals were selected into four groups. Groups 1 and 2 were pre-treated with normal saline (10 mL/kg i.p) for 28 days. Groups 3 and 4 received GBS (50 mg/kg, i.p) for 28 days. However, groups 2 and 4 were administered ISO (150 mg/kg, s.c) on days 27 and 28 respectively. Following the end of 28-day experiment, animals were euthanised and serum as well as heart tissue were harvested and processed for biochemical and histological examinations. ResultsOur results showed that ISO induced cardiac necrosis, evidenced by increased levels of cardiac injury enzymes (LDH, GGT, CK-MB and cT-1), oxidative stress markers such as GSH, SOD, and CAT levels were significantly reduced while MDA and nitrite concntration were significantly increased. Inflammatory cytokines (IL-6 and TNF-α) were significantly increased while decreasing Bcl-2, mTOR and ERK1/2 cardiac immuno-expressions. Moreso, histological investigation revealed degeneration of cardiomyofibres together with patchy necrosis. However, pre-treatment with GBS significantly inhibited apoptotic processes evidenced by increased antioxidant-mediated activation of Bcl-2, mTOR and ERK1/2. GBS also suppressed the release of cardiac injury enzymes and histological changes were significantly improved. ConclusionGBS protects myocardium against ISO-induced infarction by mechanisms related to antioxidant-mediated by enhancement of Bcl-2/mTOR/ERK1/2 activities.

Breast cancer, alcohol, and phosphate toxicity.

Alcohol consumption is associated with an increased risk of breast cancer, even at low alcohol intake levels, but public awareness of the breast cancer risk associated with alcohol intake is low. Furthermore, the causative mechanisms underlying alcohol's association with breast cancer are unknown. The present theoretical paper uses a modified grounded theory method to review the research literature and propose that alcohol's association with breast cancer is mediated by phosphate toxicity, the accumulation of excess inorganic phosphate in body tissue. Serum levels of inorganic phosphate are regulated through a network of hormones released from the bone, kidneys, parathyroid glands, and intestines. Alcohol burdens renal function, which may disturb the regulation of inorganic phosphate, impair phosphate excretion, and increase phosphate toxicity. In addition to causing cellular dehydration, alcohol is an etiologic factor in nontraumatic rhabdomyolysis, which ruptures cell membranes and releases inorganic phosphate into the serum, leading to hyperphosphatemia. Phosphate toxicity is also associated with tumorigenesis, as high levels of inorganic phosphate within the tumor microenvironment activate cell signaling pathways and promote cancer cell growth. Furthermore, phosphate toxicity potentially links cancer and kidney disease in onco-nephrology. Insights into the mediating role of phosphate toxicity may lead to future research and interventions that raise public health awareness of breast cancer risk and alcohol consumption.

Abstract 2560: Post-translational modification of focal adhesion kinase (FAK) protein increases in breast cancer cells upon exposure to conditioned medium derived from in vitro and in vivo adipocytes

Abstract Under obese conditions, dysregulation of the normal secretion of adipokines from fat cells has been shown to drive cancer progression. However, the role adipokine dysregulation plays in driving metastasis is still unclear. Recent studies have reported the activation of metastasis-associated cell signaling pathways. One such pathway involves Focal Adhesion Kinase (FAK), a non-receptor cytoplasmic protein that acts as a signal transducer between receptor activation by adipokines and downstream effector proteins that control the biological phenomenon of metastasis. Nevertheless, the association between obesity-induced changes in adipocyte secretory factors and the subsequent increase in the transactivation of FAK through phosphorylation is unknown. To elucidate the potential relationship of adipocyte-secreted factors towards activating FAK, breast cancer cells with low (MCF7) and high (MDA-MB-231) metastatic potential were exposed to in vitro and in vivo, murine adipocyte-derived, conditioned medium. For the in vitro adipocyte model, 3T3-L1 MBX cells differentiated to obesity, with fatty acids supplementation, were used. For the in vivo model, adipocyte-secreted factors from organ-cultured visceral fat harvested from C3H/HeJ female mice fed an either high fat (46%) or control (11%) diets, were collected in the medium. Adipocyte secretions from the in vitro and in vivo adipocyte samples were determined using a mouse adipokine proteomic profiler array and ELISA kit. Under obese conditions, both the 3T3-L1 MBX cells and C3H/HeJ visceral fat were found to have significant changes in their adipokine secretions in comparison to controls. Results showed that the breast cancer cells got increased FAK autophosphorylation when exposed to either in vitro or in vivo-derived obese adipocyte-conditioned medium in comparison to the untreated breast cancer cells. Citation Format: Noshin Mubtasim, Caleb Boren, Benjamin Barr, Lauren Gollahon. Post-translational modification of focal adhesion kinase (FAK) protein increases in breast cancer cells upon exposure to conditioned medium derived from in vitro and in vivo adipocytes [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2560.

Monkeypox and extracellular vesicles interlink: an unexplored chapter.

Dear Editor, Monkeypox is an infectious zoonotic disease which is caused by monkeypox virus. It belongs to the genus Orthopoxvirus and the family Poxviridae. The lethality rate of this disease ranges from 1 to 10%. The first occurrence of this disease was in the Democratic Republic of the Congo, Africa. Monkeypox spreads mainly through direct contact with body fluids, respiratory droplets of infected patients and through animals who act as its reservoir The genome of monkeypox virus is linear, enveloped, and double-stranded DNA1,2. This disease mainly has two phases, the first one is the invasive phase which is characterized mainly by fatigue, sore throat, fever, and myalgia. The second phase is the eruptive phase which is characterized by the clinical symptoms of pustular lesions on the legs, multiple lesions on the penile sulcus and huge ulcerative lesions on the perianal skin3. In the current decade, extracellular vesicles (EVs) have gained much attention due to their ability that leads to intercellular communication by activation of various cell signalling pathways. Tetraspanins are the group of surface EVs protein which involved viral particles associated with targeting cells and fission with the recipient cells via cellular membranes. Virus-derived EVs internal cargo influence the recipient cell activity. Envelop virus and EVs have biologically active cargo transportation ability4. EVs are released for normal or pathological conditions (it may be cancer cells or viral-infected cells). The origin of EVs is different as exosomes develop from the plasma membrane and apoptotic bodies develop during the apoptosis process, microvesicles secrete from the plasma membrane and endoplasmic reticulum (these are three major subpopulations of EVs). EVs regulate host–pathogen cellular communication. Viral-infected cell-derived EVs mediated protein and nucleic acid (DNA, RNA) transportation promote the viral pathogenesis progression5,6. In the context of viral infections, the biological role of EVs might have two opposing effects. EVs can influence recipient cells based boosting viral reproduction on the one hand, or they can prevent viral replication by inducing host immune responses on the other7. In addition, integrating viral proteins can cause nonparticipating immune cells to die, which would contribute to the significant loss of immune cells during the initial phases of viral infection or when the viral load is low. In addition to making immune cells more Susceptible to viral infection, intercellular transfer of viral proteins and viral cell surface receptors by EVs aids in evading the host’s immunological response by reducing lymphocyte synthesis of antibodies8. The aim of the article is to highlighted the EVs’ interrelation with the virus and encourage large-scale virologists to decode the virus and EVs related to several aspect. Ethical approval None. Sources of funding None. Authors’ contribution D.M., B.B., R.D., S.N., and P.S.: conceptualization, study design, and writing. A.D. and S.K.J.: editing and reviewing. Conflicts of interest disclosure The authors declare that they have no financial conflict of interest with regard to the content of this report. Research registration unique identifying number (UIN) None. Guarantor Saurabh K. Jha.

Emerging Role of Plant-Based Dietary Components in Post-Translational Modifications Associated with Colorectal Cancer.

Colorectal cancer (CRC) is one of the most common cancers worldwide. Its main modifiable risk factors are diet, alcohol consumption, and smoking. Thus, the right approach through lifestyle changes may lead to its prevention. In fact, some natural dietary components have exhibited chemopreventive activity through modulation of cellular processes involved in CRC development. Although cancer is a multi-factorial process, the study of post-translational modifications (PTMs) of proteins associated with CRC has recently gained interest, as inappropriate modification is closely related to the activation of cell signalling pathways involved in carcinogenesis. Therefore, this review aimed to collect the main PTMs associated with CRC, analyse the relationship between different proteins that are susceptible to inappropriate PTMs, and review the available scientific literature on the role of plant-based dietary compounds in modulating CRC-associated PTMs. In summary, this review suggested that some plant-based dietary components such as phenols, flavonoids, lignans, terpenoids, and alkaloids may be able to correct the inappropriate PTMs associated with CRC and promote apoptosis in tumour cells.

SOX2-high cancer cells exhibit an aggressive phenotype, with increases in stemness, proliferation and invasion, as well as higher metabolic activity and ATP production.

Cancer stem cells (CSCs) are responsible for cancer recurrence, treatment failure and metastatic dissemination. As such, the elimination of CSCs represents one of the most important approaches for the future of cancer treatment. Among other properties, CSCs show the activation of particular cell signalling pathways and the over-expression of certain transcription factors, such as SOX2. Herein, we describe a new model system to isolate stem-like cancer cells, based on the functional transcriptional activity of SOX2. Briefly, we employed a SOX2-enhancer-GFP-reporter system to isolate cancer cells with high SOX2 transcriptional activity by FACS sorting. The over-expression of SOX2 in this sub-population was validated by Western blot analysis and flow cytometry. SOX2-high cancer cells showed CSCs features, such as greater mammosphere forming ability, validating that this sub-population was enriched in CSCs. To further explore the model, we analysed other stemness characteristics in MCF7 and MDA-MB-231 breast cancer cell lines, corroborating that SOX2-high cells were more metabolically active, proliferative, migratory, invasive, and drug-resistant. SOX2-high MDA-MB-231 cells also showed a loss of E-cadherin expression, and increased Vimentin expression, consistent with an epithelial-mesenchymal transition (EMT). Therefore, endogenous SOX2 transcriptional activity and protein levels are mechanistically linked to aggressive phenotypic behaviours and energy production in CSCs.

Resistance Exercise-Induced Increases in Muscle Myostatin mRNA and Protein Expression Are Subsequently Decreased in Circulation in the Presence of Increased Levels of the Extracellular Matrix Stabilizing Protein Decorin.

Resistance exercise (RE) activates cell signaling pathways associated with myostatin. Decorin is located in the extracellular matrix (ECM) and can block the inhibitory effect of myostatin. This study sought to determine the impact of low-load (LL) and high-load (HL) RE on myostatin mRNA and protein expression along with changes in muscle decorin and circulating follistatin. Ten resistance-trained men performed a LL (50% 1RM) and HL (80% 1RM) RE session using the angled leg press and leg extension with load and volume equated. Venous blood samples and muscle biopsies were obtained prior to and at 3h and 24h following each RE session. Muscle myostatin mRNA expression was increased at 24h post-exercise (p = 0.032) in LL and at 3h (p = 0.044) and 24h (p = 0.003) post-exercise in HL. Muscle decorin was increased at 24h post-exercise (p < 0.001) in LL and HL; however, muscle myostatin was increased at 24h post-exercise (p < 0.001) only in HL. For muscle Smad 2/3, no significant differences were observed (p > 0.05). Serum follistatin was increased and myostatin decreased at 24h post-exercise (p < 0.001) in LL and HL. Muscle myostatin gene and protein expression increased in response to HL RE. However, serum myostatin was decreased in the presence of increases in decorin in muscle and follistatin in circulation. Therefore, our data suggest a possible mechanism may exist where decorin within the ECM is able to bind to, and decrease, myostatin that might otherwise enter the circulation for activin IIB (ACTIIB) receptor binding and subsequent canonical signaling through Smad 2/3.

MiRNA-223-3p regulates ECT2 to promote proliferation, invasion, and metastasis of gastric cancer through the Wnt/β-catenin signaling pathway.

Expression of the guanine nucleotide exchange factor epithelial cell transforming 2 (ECT2) is elevated in gastric cancer (GC) but its biological function in GC is poorly understood. MicroRNAs (miRNAs) have great potential as therapeutic targets for GC through their ability to modulate gene expression. In the present study, we sought to identify potential miRNA-mRNA-protein regulatory pathways that might control ECT2 expression and function in GC. ECT2 expression was examined in clinical GC specimens by immunohistochemical staining, and protein levels were correlated with clinicopathological features and prognosis. TargetScan was used to identify potential ECT2 mRNA-complementary miRNAs, and the roles of ECT2 and miRNA-223-3p (miR-223-3p) in GC cell biology and signaling pathway activation were examined by targeted knockdown (KD) or overexpression (OE) of ECT2 and miR-223-3p in GC cell lines. A murine GC xenograft model was developed to explore the impact of ECT2 OE on tumor growth in vivo. ECT2 expression was significantly elevated in GC specimens compared with normal gastric tissues and the level correlated positively with depth of invasion, ulceration, vascular tumor thrombus, neural invasion, and lymph node metastasis (p < 0.05). ECT2 was an independent prognostic factor for overall survival of GC patients (high ECT2 expression v.s. low ECT2 expression: χ2 = 29.831, p < 0.001). ECT2 KD or miR-223-3p OE markedly suppressed the proliferation, migration, and invasion of GC cells in vitro, whereas ECT2 OE had the opposite effects. ECT2 OE also promoted the growth of GC tumors in vivo. Tumor expression of Wnt2, β-catenin, and several downstream target proteins in GC cells were decreased by ECT2 KD or miR-223-3p OE but increased by ECT2 OE. miR-223-3p regulates ECT2 expression to promote tumorigenic behavior of GC via activation of the Wnt/β-catenin signaling pathway, suggesting that ECT2 and miR-223-3p as potential therapeutic targets for GC.

Apigenin in cancer therapy: From mechanism of action to nano-therapeutic agent.

Apigenin (APG) is a flavonoid presence in beverages, vegetables, and fruits containing anti-diabetic, anti-oxidant, and anti-viral activities, as well as cancer management properties. There is growing evidence that APG presented extensive anti-cancer effects in several cancer types by modulating various cellular processes, including angiogenesis, apoptosis, metastasis, autophagy, cell cycle, and immune responses, through activation or inhibition of different cell signaling pathways and molecules. By emerging nanotechnology and its advent in the biomedicine field, cancer therapy has been changed based on nanotechnology-based delivery systems. APG nanoformulations have been used to target tumor cells specifically, improve cellular uptake of APG, and overcome limitations of the free form of APG, such as low solubility and poor bioavailability. In this review, the biotherapeutic activity of APG and its mechanisms, both in free form and nanoformulation, toward cancer cells are discussed to shed some light on APG anti-tumor activity in different cancers.

Stem Cell Models for Cancer Therapy.

Metastatic progression of female breast and colon cancer represents a major cause of mortality in women. Spontaneous/acquired resistance to conventional and targeted chemo-endocrine therapy is associated with the emergence of drug-resistant tumor-initiating cancer stem cell populations. The cancer-initiating premalignant stem cells exhibit activation of select cancer cell signaling pathways and undergo epithelial–mesenchymal transition, leading to the evolution of a metastatic phenotype. The development of reliable cancer stem cell models provides valuable experimental approaches to identify novel testable therapeutic alternatives for therapy-resistant cancer. Drug-resistant stem cell models for molecular subtypes of clinical breast cancer and for genetically predisposed colon cancer are developed by selecting epithelial cells that survive in the presence of cytostatic concentrations of relevant therapeutic agents. These putative stem cells are characterized by the expression status of select cellular and molecular stem cell markers. The stem cell models are utilized as experimental approaches to examine the stem-cell-targeted growth inhibitory efficacy of naturally occurring dietary phytochemicals. The present review provides a systematic discussion on (i) conceptual and experimental aspects relevant to the chemo-endocrine therapy of breast and colon cancer, (ii) molecular/cellular aspects of cancer stem cells and (iii) potential stem-cell-targeting lead compounds as testable alternatives against the progression of therapy-resistant breast and colon cancer.

Apoptosis and (in) Pain-Potential Clinical Implications.

The deregulation of apoptosis is involved in the development of several pathologies, and recent evidence suggests that apoptosis may be involved in chronic pain, namely in neuropathic pain. Neuropathic pain is a chronic pain state caused by primary damage or dysfunction of the nervous system; however, the details of the molecular mechanisms have not yet been fully elucidated. Recently, it was found that nerve endings contain transient receptor potential (TRP) channels that sense and detect signals released by injured tissues and respond to these damage signals. TRP channels are similar to the voltage-gated potassium channels or nucleotide-gated channels that participate in calcium and magnesium homeostasis. TRP channels allowing calcium to penetrate into nerve terminals can activate apoptosis, leading to nerve terminal destruction. Further, some TRPs are activated by acid and reactive oxygen species (ROS). ROS are mainly produced in the mitochondrial respiratory chain, and an increase in ROS production and/or a decrease in the antioxidant network may induce oxidative stress (OS). Depending on the OS levels, they can promote cellular proliferation and/or cell degeneration or death. Previous studies have indicated that proinflammatory cytokines, such as tumor necrosis factor-α (TNF-α), play an important role in the peripheral mediation of neuropathic pain. This article aims to perform a review of the involvement of apoptosis in pain, particularly the role of OS and neuroinflammation, and the clinical relevance of this knowledge. The potential discovery of new biomarkers and therapeutic targets can result in the development of more effective and targeted drugs to treat chronic pain, namely neuropathic pain. Highlights: Oxidative stress and neuroinflammation can activate cell signaling pathways that can lead to nerve terminal destruction by apoptosis. These could constitute potential new pain biomarkers and targets for therapy in neuropathic pain.