Growth Factor Stimulation Research Articles

Abstract PS18-03: Differential gene expression in luminal-type invasive lobular carcinoma and invasive ductal carcinoma by MammaPrint risk stratification

Abstract Background: Invasive lobular carcinoma (ILC) comprises 10-15% of breast tumors and is the second most common histological type after invasive ductal carcinoma (IDC). Patients with ILC are often diagnosed at an older age and more advanced stage than those with IDC. Late recurrences and worse long-term survival suggest the need for improved approaches to treatment optimization and exploration of molecular pathways unique to ILC. Although previous reports have described comprehensive transcriptomic profiling of ILC, these were limited by small sample sizes. Furthermore, differential gene expression between ILC and IDC within genomic risk groups and molecular subtypes has yet to be explored. Here we characterize differential gene expression between ILC and IDC in a large, age-matched patient subset categorized by 70-gene signature/MammaPrint (MP) risk and 80-gene signature/BluePrint (BP) subtype. Methods: The prospective FLEX Registry (NCT03053193) includes stage I-III primary invasive breast cancer patients who receive MP/BP testing and consent to full transcriptome and clinical data collection. This sub-analysis included 450 ILC patients enrolled from 2017 to present. Compared with a random selection of IDC patients (n=450, mean age, 60 years), ILC patients were older (mean, 63 years, p<0.001). Thus, we selected an age-matched subset for differential gene expression analysis. There were few non-Luminal ILCs; thus, gene expression analyses were limited to BP Luminal tumors. A subset of 413 age-matched pairs (n=826) of ILC and IDC were used for analysis. Gene expression data were quantile normalized using R limma package, and differentially expressed genes (DEGs) were compared between groups. DEGs with an adjusted p<0.05 and log2 fold change > ± 1.0 were considered significant. Results: ILC represented 13% of FLEX cases (n=450/3562), and were 81% lymph node-negative, 99% ER+, 94% HER2-negative, and 68% MP Low Risk (LR). By BP, ILC were 99% Luminal, 1% HER2, and <1% Basal type. BP Luminal ILC were predominantly grade 2 (63%), T1 (61%), node-negative (84%), and MP LR (69%). Menopausal status, nodal status, ethnicity, BMI distribution, and frequency of type 2 diabetes mellitus were similar between ILC and IDC. However, IDC were more likely to be MP HR (46% IDC vs. 31% ILC, p<0.001) and grade 3 (15% IDC vs. 4% ILC, p<0.001). ILC were more likely to be T3 (10% ILC vs. 1% IDC, p<0.001). We found 4 DEGs common to all comparisons: all Luminal ILC vs. IDC, MP LR ILC vs. IDC, and MP HR ILC vs. IDC. ILC had lower expression of CDH1 (E-cadherin) than IDC, regardless of MP risk. Including CDH1, 6 unique genes were differentially expressed in LR ILC compared with IDC, and 21 genes were differentially expressed in HR ILC compared with IDC. Genes with increased expression in HR ILC were related to immune cell migration/chemotaxis, hormone signaling, and growth factor signaling. HR ILCs were also enriched for TGFβ signaling and angiogenesis pathway genes. Conclusions: Here we report differential clinical and molecular characteristics between ILC and IDC in a large, age-matched patient subset. Regardless of MP risk, expression of CDH1 was lower in ILC compared with IDC. Approximately one-third of ILCs were MP HR, and we report a greater number and diversity of DEGs between HR ILC and HR IDC compared with LR tumors, in particular genes related to TGFβ signaling. TGFβ pathway genes play a variety of roles in the tumor microenvironment, including induction of angiogenesis, fibroblast growth factor stimulation, and inhibition and/or exclusion of an immune response. These results suggest that therapeutic strategies targeting the TGFβ pathway may be future avenues of exploration in ILC, although further studies are warranted to characterize underlying molecular mechanisms. Citation Format: Beth-Ann Lesnikoski, Jennifer A. Crozier, Gordan Srkalovic, Patricia Robinson, Clodia Osipo, Kaylan Banda, Heather M. Kling, Josien Haan, William Audeh, FLEX Investigators Group. Differential gene expression in luminal-type invasive lobular carcinoma and invasive ductal carcinoma by MammaPrint risk stratification [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PS18-03.

Growth factor stimulation for the healing of traumatic ulcers with liquid rice hull smoke

ObjectiveThe healing process of a traumatic ulcer requires growth factors to rebuild the lost tissue after the inflammatory process has been completed. Liquid rice hull smoke (LR-HS) has shown unique anti-inflammatory properties. This study analyses the role of LR-HS in growth factor stimulation for the healing of traumatic ulcers, such as fibroblast growth factor (FGF), vascular endothelial growth factor (VEGF), platelet-derived growth factor (PDGF), and collagen type 1 (COL-1) expression. MethodsWe obtained LR-HS from the pyrolysis of rice hulls. Traumatic ulcers were created in the labial fornix incisive inferior of Wistar rats and treated with LR-HS once a day for 3, 5, and 7 days. The control group was treated with sterile water. Each animal was sacrificed after treatment, and its labial fornix incisive inferior tissues were biopsied and immunohistochemically stained to examine FGF, VEGF, PDGF, and COL-1 expression. ResultThe treatment of traumatic ulcers with LR-HS showed an increase in FGF, VEGF, PDGF, and COL-1 expression. VEGF expression increased under LR-HS treatment compared with the control 7-day treatment groups (p < 0.000). FGF and COL-1 expression increased under LR-HS treatment compared with the control 5- and 7-day treatment groups (p < 0.000). PDGF expression increased after treatment with LR-HS for 3, 5, and 7 days (p < 0.000). ConclusionThis study has demonstrated that LR-HS can induce the expression of growth factors during the healing of a traumatic ulcer using immunohistochemical staining. We suggest that LR-HS can be used as a herbal medicine for oral ulcer therapy.

HOMER3 facilitates growth factor-mediated \u03b2-Catenin tyrosine phosphorylation and activation to promote metastasis in triple negative breast cancer

BackgroundHOMER family scaffolding proteins (HOMER1-3) play critical roles in the development and progression of human disease by regulating the assembly of signal transduction complexes in response to extrinsic stimuli. However, the role of HOMER protein in breast cancer remains unclear.MethodsHOMER3 expression was examined by immunohistochemistry in breast cancer patient specimens, and its significance in prognosis was assessed by Kaplan–Meier survival analysis. The effects of HOMER3 in growth factor-induced β-Catenin activation were analyzed by assays such as TOP/FOP flash reporter, tyrosine phosphorylation assay and reciprocal immunoprecipitation (IP) assay. Role of HOMER3 in breast cancer metastasis was determined by cell function assays and mice tumor models.ResultsHerein, we find that, among the three HOMER proteins, HOMER3 is selectively overexpressed in the most aggressive triple negative breast cancer (TNBC) subtype, and significantly correlates with earlier tumor metastasis and shorter patient survival. Mechanismly, HOMER3 interacts with both c-Src and β-Catenin, thus providing a scaffolding platform to facilitate c-Src-induced β-Catenin tyrosine phosphorylation under growth factor stimulation. HOMER3 promotes β-Catenin nuclear translocation and activation, and this axis is clinically relevant. HOMER3 promotes and is essential for EGF-induced aggressiveness and metastasis of TNBC cells both in vitro and in vivo.ConclusionThese findings identify a novel role of HOMER3 in the transduction of growth factor-mediated β-Catenin activation and suggest that HOMER3 might be a targetable vulnerability of TNBC.

Master Regulators of Muscle Atrophy: Role of Costamere Components.

The loss of muscle mass and force characterizes muscle atrophy in several different conditions, which share the expression of atrogenes and the activation of their transcriptional regulators. However, attempts to antagonize muscle atrophy development in different experimental contexts by targeting contributors to the atrogene pathway showed partial effects in most cases. Other master regulators might independently contribute to muscle atrophy, as suggested by our recent evidence about the co-requirement of the muscle-specific chaperone protein melusin to inhibit unloading muscle atrophy development. Furthermore, melusin and other muscle mass regulators, such as nNOS, belong to costameres, the macromolecular complexes that connect sarcolemma to myofibrils and to the extracellular matrix, in correspondence with specific sarcomeric sites. Costameres sense a mechanical load and transduce it both as lateral force and biochemical signals. Recent evidence further broadens this classic view, by revealing the crucial participation of costameres in a sarcolemmal “signaling hub” integrating mechanical and humoral stimuli, where mechanical signals are coupled with insulin and/or insulin-like growth factor stimulation to regulate muscle mass. Therefore, this review aims to enucleate available evidence concerning the early involvement of costamere components and additional putative master regulators in the development of major types of muscle atrophy.

Global Histone H3 Lysine 4 Trimethylation (H3K4me3) Landscape Changes in Response to TGFβ.

TGFβ expression acts as a biomarker of poor prognosis in prostate cancer. It plays a dual functional role in prostate cancer. In the early stages of the tumor, it acts as a tumor suppressor while at the later stages of tumor development, it promotes metastasis. The molecular mechanisms of action of TGFβ are largely understood through the canonical and non-canonical signal transduction pathways. Our understanding of the mechanisms that establish transient TGFβ stimulation into stable gene expression patterns remains incomplete. Epigenetic marks like histone H3 modifications are directly linked with gene expression and they play an important role in tumorigenesis. In this report, we performed chromatin immunoprecipitation-sequencing (ChIP-Seq) to identify the genome-wide regions that undergo changes in histone H3 Lysine 4 trimethylation (H3K4me3) occupancy in response to TGFβ stimulation. We also show that TGFβ stimulation can induce acute epigenetic changes through the modulation of H3K4me3 signals at genes belonging to special functional categories in prostate cancer. TGFβ induces the H3K4me3 on its own ligands like TGFβ, GDF1, INHBB, GDF3, GDF6, BMP5 suggesting a positive feedback loop. The majority of genes were found to be involved in the positive regulation of transcription from the RNA polymerase II promoter in response to TGFβ. Other functional categories were intracellular protein transport, brain development, EMT, angiogenesis, antigen processing, antigen presentation via MHC class II, lipid transport, embryo development, histone H4 acetylation, positive regulation of cell cycle arrest, and genes involved in mitotic G2 DNA damage checkpoints. Our results link TGFβ stimulation to acute changes in gene expression through an epigenetic mechanism. These findings have broader implications on epigenetic bases of acute gene expression changes caused by growth factor stimulation.

Dehydrated Human Amnion-Chorion Product as an Adjunct to Scaling and Root Planing inMaintenance Patients: A Pilot Study

Background: Adjunctive therapies are often combined with Scaling and Root Planing (SRP) procedures to limit bacterial recolonization of periodontal pockets and to modulate the wound healing environment. While local delivery antimicrobial therapies are effective, they do not promote soft tissue repair at these sites. BioXclude, a dehydrated human de-epithelialized amnion-chorion membrane product (ddACM) sourced from donated human placental tissue, has been shown to have antimicrobial properties, contain stimulatory growth factors known to activate wound healing processes, and possess critical attachment substrates for cellular adhesion and migration. Given this combination, it was hypothesized that ddACM would be an effective adjunctive therapy for the treatment of periodontal pockets following SRP procedures. Methods: Sixteen non-smoking patients, compliant with a regular periodontal maintenance program, were included in this study. A total of 30 sites showing a ≥ 2 mm increase in periodontal probing depth (PPD), with associated bleeding on probing (BOP), were chosen for treatment. Following standard SRP procedures with local anesthesia, ddACM was placed into the treated area. A six-week periodontal re-evaluation was completed, with the primary outcome variable being PD reduction and secondary outcomes that included resolution of bleeding on probing (BOP). Results: Mean PD reduction was 2.55 mm ± 0.34 mm, with three sites showing ≥ 4 mm ± 0.2 mm reduction in PD. BOP was resolved in 83% of the treated sites. Conclusion: ddACM provides significant benefit as an adjunctive therapy to SRP for the treatment of periodontal pockets in terms of PD reduction and resolution of BOP.

Roles of growth factors on vascular remodeling in pulmonary hypertension

Pulmonary hypertension (PH) is defined as mean pulmonary arterial pressure at rest ≥25 mmHg. Pulmonary arterial hypertension (PAH) is classified as group 1 of PH and is a progressive and fatal disease of the pulmonary artery. The pathogenesis is sustained pulmonary vasoconstriction and pulmonary vascular remodeling, which cause progressive elevations in pulmonary vascular resistance and pulmonary arterial pressure. Elevated pulmonary arterial pressure leads to right heart failure and finally death. The pulmonary vascular remodeling is triggered by an increase in cytosolic Ca2+ concentration ([Ca2+]cyt). [Ca2+]cyt is regulated by the stimulation of vasoconstrictors and growth factors though their receptors and ion channels on the plasma membrane. It has been reported that the epidermal growth factor (EGF), fibroblast growth factor (FGF), insulin-like growth factor (IGF), vascular endothelial growth factor (VEGF), and platelet-derived growth factor (PDGF) are involved in the development of PAH. Upon binding of these growth factors with their specific receptor tyrosine kinases, their receptors activate cytosolic Ca2+ signaling and signal transduction cascades to induce cell proliferation, differentiation, and migration. Expressions of some growth factors and their receptors upregulate in PAH patients, which contributes to the formation of vascular remodeling and plexiform lesions in PAH. We have recently found that enhanced Ca2+-sensing receptor (CaSR) function is involved the development of PAH and CaSR expression is upregulated by PDGF in pulmonary arterial smooth muscle cells (PASMCs) from idiopathic PAH patients. This review will be discussed the physiological and pathological roles of growth factors in PAH.

MicroRNA-146a inhibition promotes total neurite outgrowth and suppresses cell apoptosis, inflammation, and STAT1/MYC pathway in PC12 and cortical neuron cellular Alzheimer's disease models.

This study aimed to explore the effect of microRNA (miR)-146a inhibition on regulating cell apoptosis, total neurite outgrowth, inflammation, and STAT1/MYC pathway in Alzheimer's disease (AD). PC12 and cortical neuron cellular AD models were constructed by Aβ1-42 insult. For the former model, nerve growth factor (NGF) stimulation was previously conducted. miR-146a inhibitor and negative-control (NC) inhibitor were transfected into the two cellular AD models, and then cells were named miR-inhibitor group and NC-inhibitor group, respectively. After transfection, cell apoptosis, total neurite outgrowth, supernatant inflammation cytokines, and STAT1/MYC pathway were detected. miR-146a expression was similar between PC12 cellular AD model and control cells (NGF-stimulated PC12 cells), while miR-146a expression was increased in cortical neuron cellular AD model compared with control cells (rat embryo primary cortical neurons). In both PC12 and cortical neuron cellular AD models, miR-146a expression was reduced in miR-inhibitor group compared with NC-inhibitor group after transfection. Furthermore, cell apoptosis was attenuated, while total neurite outgrowth was elevated in miR-inhibitor group compared with NC-inhibitor group. As for supernatant inflammatory cytokines, tumor necrosis factor-α, interleukin (IL)-1β, IL-6, and IL-17 levels were lower in miR-inhibitor group than in NC-inhibitor group. Additionally, STAT1 and c-Myc mRNA and protein expressions were attenuated in miR-inhibitor group compared with NC-inhibitor group. In conclusion, miR-146a potentially represented a viable therapeutic target for AD.

Клиническое значение определения биомаркера крови ST2 у больных с хронической сердечной недостаточностью

Stimulating growth factor expressed by gene 2 (ST2) and natriuretic peptides (BNP and NT-proBNP) reflect the flow of two different, but intersecting biological processes, so the markers provide independent and complementary information on the patient’s condition. Being markers of hemodynamic instability or stretching of cardiomyocytes NT-proBNP/BNP is more suitable for detecting chronic heart failure, but this is not enough to rehospitalize patients for 30 days with the predictive qualities of this marker. ST2 is the strongest and clinically significant prognostic marker of cumulative cardiovascular events and mortality. The use of new highly specific cardiobiomarkers will make it possible to predict the development of cardiovascular complications with a high degree of predictive value, and therefore will allow prophylactically prescribing appropriate pathogenetic therapy aimed at improving the quality of life of patients, reducing mortality from cardiovascular diseases and reducing the economic costs of the state for treating heart failure and cardiovascular complications.

In memory of Academician A.I. Vorob'ev, Russian Academy of Medical Sciences and Russian Academy of Sciences Stability of Granulopoiesis Chronology under R(G)-DHAP Immuno-chemotherapy-Induced Cytotoxic Stress in Non-Hodgkin’s Lymphomas

Background. Chronology of granulopoiesis based on periodic hematopoiesis model has been thoroughly studied. However, the pattern of influence of immunotherapy-induced cytotoxic stress on the biological rhythm of a stem cell development requires further investigation. The interaction of antitumor drugs with normal hematopoietic cells is relevant for assessing the intensity of chemotherapy adverse events. Besides, there is a demand for studying hematopoiesis under cytotoxic stress to predict immunological reactivity as a condition for efficacy of immunotherapeutic agents, the effect of which is based on cell immunity. Aim. To study the chronological pattern of leukocyte count dynamics after R(G)-DHAP immunochemotherapy in non-Hodgkin’s lymphomas. Materials &amp; Methods. The dynamics of leukocyte count changes after R(G)-DHAP immunochemotherapy was analyzed using the data of 39 treatment courses in 19 non-Hodgkin’s lymphomas patients. After 18 out of 39 cycles of treatment granulocyte colony-stimulating factor (G-CSF) was administered to prevent granulocytopenia, in other cases the previously planned hematopoietic stem cell mobilization was performed according to the accepted protocol. Results. Time to activation of spontaneous granulopoiesis depends neither on G-CSF stimulation, nor on the total dose of growth-stimulating factor and corresponds on average to Day 10 or Day 11 of the break from the last day of immunochemotherapy. The tendency of shorter agranulocytosis duration on prophylactic use of G-CSF is associated with transient hyperleukocytosis at an early stage after completing immunochemotherapy. Regimens with platinum-based drugs, like R(G)-DHAP, are suggested to be combined with immunochemotherapeutic agents in patients with the failure of first-line chemotherapy. The time interval preceding myelopoiesis activation within the first days of the break between the courses is likely to contribute to the initiation of treatment with immunotherapeutic drugs after second-line chemotherapy. Conclusion. The determination of granulopoiesis dynamics under R(G)-DHAP immunochemotherapy-induced cytotoxic stress enables to plan the optimum G-CSF regimen and to predict the optimum timing of immune antitumor effect combined with chemotherapy.

Излечение мышей с распространённой аденокарциномой яичников СаО1 с помощью белков сыворотки крови

After removal of the primary tumor node, tumor-specific activity appears in the serum that blocks tumor growth in mice. This activity is observed at the time interval when activity of the tumor growth-stimulating factor is not determined. Administration of blood serum (0.1 ml) from mice with removed tumor to mice with CaO1 adenocarcinoma for 14 days led first to a stop of its growth, and then to tumor regression. The animals cured of adenocarcinoma lived for at least one year without signs of relapse. The cured animals did not develop resistance to repeated tumor transplantation. Repeated transplantation led to the growth of the new tumor. No cellular immune response was observed on histological slides of the regressing tumor. It was concluded that a serum factor is required for the growth of a tumor in the body and the state of the serum with blocked activity of this tumor-stimulating factor can be used for tumor treatment in oncology patients. This is the first result in the syngeneic system, when the tumor was cured by syngeneic serum proteins.

ASSESSING THE UTILITY OF A CYTOKINE IN THE DIAGNOSIS OF CERVICAL INTRAEPITHELIAL NEOPLASM AMONG WOMEN IN BENIN CITY, NIGERIA

Cervical cancer has been recognized as the fourth commonest cancer worldwide with 70 % of deaths from cases occurring from developing countries. The mortality rate among women in Nigeria can be drastically reduced by early specific diagnosis of premalignant lesions. Although Cytokines are not routinely used as diagnostic markers for cervical cancer, this study investigated the strength of Granulocyte Monocyte Colony Stimulating growth Factor (GM-CSF) as an indicator in the prediction of Cervical Intraepithelial Neoplasm (CIN). The study was carried out at the Department of Obstetrics and Gynaecology, University of Benin Teaching Hospital Benin City and Department of Chemical Pathology, Edo University Iyamho, Edo state Nigeria, between August, 2017 and January 2019. It was a cross-sectional prospective study. A total of 197 female participants were recruited for this study. Based on histological diagnosis they were grouped as; Negative, CIN 1, CIN 2 and CIN 3. Venous blood was obtained from participants and serum GM-CSF levels were determined using standardized laboratory methods. The results showed a statistically significant elevation of serum GM-CSF in participants with CIN lesions (56.27±1.21ng/ml) compared to those without (33.43±3.03ng/ml). The study also suggests a 56.3% sensitivity and a 95.3% specificity of GM-CSF marker to CIN with an Area under the Curve (AUC) of 0 .907, p&lt;0.001 from the Receivers Operator Curve (ROC). This research suggests the use of GM-CSF as a good marker for screening of those at risk of cervical cancer because of its high prospect to reduce the incidence of false positives. However, because of the low sensitivity, it is recommended that GM-CSF be used as a CIN diagnostic tool in combination with the standard Pap smear test.

Biomarkers of acute myocardial infarction: diagnostic and prognostic value. Part 2 (Literature review)

In the second part of the review, we continue the discussion of biomarkers that have a diagnostic and prognostic significance in acute myocardial infarction (AMI). The study of the AMI pathophysiology through the experimental and clinical research contributes to the discovery of new regulatory molecules and pathogenetic mechanisms underlying AMI. At the same time, many molecules involved in the pathogenesis of AMI can be used as effective biomarkers for the diagnosis and prediction of AMI. This article discusses in detail the diagnostic and prognostic value of inflammatory biomarkers of AMI (C-reactive protein, interleukin-6, tumor necrosis factor-alpha, myeloperoxidase, matrix metalloproteinases, soluble form of CD40 ligand, procalcitonin, placental growth factor) and a number of recently discovered new biomarkers of AMI (microribonucleic acids, galectin-3, stimulating growth factor expressed by gene 2, growth differentiation factor 15, proprotein convertase of subtilisin-kexin type 9).

PTEN and Other PtdIns(3,4,5)P3 Lipid Phosphatases in Breast Cancer.

The phosphoinositide 3-kinase (PI3K)/AKT signalling pathway is hyperactivated in ~70% of breast cancers. Class I PI3K generates PtdIns(3,4,5)P3 at the plasma membrane in response to growth factor stimulation, leading to AKT activation to drive cell proliferation, survival and migration. PTEN negatively regulates PI3K/AKT signalling by dephosphorylating PtdIns(3,4,5)P3 to form PtdIns(4,5)P2. PtdIns(3,4,5)P3 can also be hydrolysed by the inositol polyphosphate 5-phosphatases (5-phosphatases) to produce PtdIns(3,4)P2. Interestingly, while PTEN is a bona fide tumour suppressor and is frequently mutated/lost in breast cancer, 5-phosphatases such as PIPP, SHIP2 and SYNJ2, have demonstrated more diverse roles in regulating mammary tumourigenesis. Reduced PIPP expression is associated with triple negative breast cancers and reduced relapse-free and overall survival. Although PIPP depletion enhances AKT phosphorylation and supports tumour growth, this also inhibits cell migration and metastasis in vivo, in a breast cancer oncogene-driven murine model. Paradoxically, SHIP2 and SYNJ2 are increased in primary breast tumours, which correlates with invasive disease and reduced survival. SHIP2 or SYNJ2 overexpression promotes breast tumourigenesis via AKT-dependent and independent mechanisms. This review will discuss how PTEN, PIPP, SHIP2 and SYNJ2 distinctly regulate multiple functional targets, and the mechanisms by which dysregulation of these distinct phosphoinositide phosphatases differentially affect breast cancer progression.

Location-specific inhibition of Akt reveals regulation of mTORC1 activity in the nucleus

The mechanistic target of rapamycin complex 1 (mTORC1) integrates growth, nutrient and energy status cues to control cell growth and metabolism. While mTORC1 activation at the lysosome is well characterized, it is not clear how this complex is regulated at other subcellular locations. Here, we combine location-selective kinase inhibition, live-cell imaging and biochemical assays to probe the regulation of growth factor-induced mTORC1 activity in the nucleus. Using a nuclear targeted Akt Substrate-based Tandem Occupancy Peptide Sponge (Akt-STOPS) that we developed for specific inhibition of Akt, a critical upstream kinase, we show that growth factor-stimulated nuclear mTORC1 activity requires nuclear Akt activity. Further mechanistic dissection suggests that nuclear Akt activity mediates growth factor-induced nuclear translocation of Raptor, a regulatory scaffolding component in mTORC1, and localization of Raptor to the nucleus results in nuclear mTORC1 activity in the absence of growth factor stimulation. Taken together, these results reveal a mode of regulation of mTORC1 that is distinct from its lysosomal activation, which controls mTORC1 activity in the nuclear compartment.

Borax induces osteogenesis by stimulating NaBC1 transporter via activation of BMP pathway

The intrinsic properties of mesenchymal stem cells (MSCs) make them ideal candidates for tissue engineering applications. Efforts have been made to control MSC behavior by using material systems to engineer synthetic extracellular matrices and/or include soluble factors in the media. This work proposes a simple approach based on ion transporter stimulation to determine stem cell fate that avoids the use of growth factors. Addition of borax alone, transported by the NaBC1-transporter, enhanced MSC adhesion and contractility, promoted osteogenesis and inhibited adipogenesis. Stimulated-NaBC1 promoted osteogenesis via the BMP canonical pathway (comprising Smad1/YAP nucleus translocation and osteopontin expression) through a mechanism that involves simultaneous NaBC1/BMPR1A and NaBC1/α5β1/αvβ3 co-localization. We describe an original function for NaBC1 transporter, besides controlling borate homeostasis, capable of stimulating growth factor receptors and fibronectin-binding integrins. Our results open up new biomaterial engineering approaches for biomedical applications by a cost-effective strategy that avoids the use of soluble growth factors.

PKM2-Induced the Phosphorylation of Histone H3 Contributes to EGF-Mediated PD-L1 Transcription in HCC.

High expression of programmed death-ligand-1 (PD-L1) in hepatocellular carcinoma (HCC) cells usually inhibits the proliferation and functions of T cells, leading to immune suppression in tumor microenvironment. However, very little has been described regarding the mechanism of PD-L1 overexpression in HCC cells. In the present study, we found epidermal growth factor (EGF) stimulation promoted the expression of PD-L1 mRNA and protein in HCC cells. Inhibition of epidermal growth factor receptor (EGFR) could reverse EGF-induced the expression of PD-L1 mRNA and protein. Subsequently, we also observed that the phosphorylation level of Pyruvate kinase isoform M2 (PKM2) at Ser37 site was also increased in response to EGF stimulation. Expression of a phosphorylation-mimic PKM2 S37D mutant stimulated PD-L1 expression as well as H3-Thr11 phosphorylation in HCC cells, while inhibition of PKM2 significantly blocked EGF-induced PD-L1 expression and H3-Thr11 phosphorylation. Furthermore, mutation of Thr11 of histone H3 into alanine abrogated EGF-induced mRNA and protein expression of PD-L1, Chromatin immunoprecipitation (ChIP) assay also suggested that EGF treatment resulted in enhanced H3-Thr11 phosphorylation at the PD-L1 promoter. In a diethylnitrosamine (DEN)-induced rat model of HCC, we found that the expression of phosphorylated EGFR, PKM2 nuclear expression, H3-Thr11 phosphorylation as well as PD-L1 mRNA and protein was higher in the livers than that in normal rat livers. Taken together, our study suggested that PKM2-dependent histone H3-Thr11 phosphorylation was crucial for EGF-induced PD-L1 expression at transcriptional level in HCC. These findings may provide an alternative target for the treatment of hepatocellular carcinoma.

Neuroprotective effects of Hericium erinaceus (Bull.: Fr.) Pers. against high-dose corticosterone-induced oxidative stress in PC-12 cells

BackgroundHericium erinaceus is a culinary and medicinal mushroom in Traditional Chinese Medicines. It has numerous pharmacological effects including immunomodulatory, anti-tumour, anti-microbial, anti-aging and stimulation of nerve growth factor (NGF) synthesis, but little is known about its potential role in negating the detrimental effects of oxidative stress in depression. The present study investigated the neuroprotective effects of H. erinaceus standardised aqueous extract (HESAE) against high-dose corticosterone-induced oxidative stress in rat pheochromocytoma (PC-12) cells, a cellular model mimicking depression.MethodsPC-12 cells was pre-treated with HESAE for 48 h followed by 400 μM corticosterone for 24 h to induce oxidative stress. Cells in complete medium without any treatment or pre-treated with 3.125 μg/mL desipramine served as the negative and positive controls, respectively. The cell viability, lactate dehydrogenase (LDH) release, endogenous antioxidant enzyme activities, aconitase activity, mitochondrial membrane potentials (MMPs), intracellular reactive oxygen species (ROS) levels and number of apoptotic nuclei were quantified. In addition, HESAE ethanol extract was separated into fractions by chromatographic methods prior to spectroscopic analysis.ResultsWe observed that PC-12 cells treated with high-dose corticosterone at 400 μM had decreased cell viability, reduced endogenous antioxidant enzyme activities, disrupted mitochondrial function, and increased oxidative stress and apoptosis. However, pre-treatment with HESAE ranging from 0.25 to 1 mg/mL had increased cell viability, decreased LDH release, enhanced endogenous antioxidant enzyme activities, restored MMP, attenuated intracellular ROS and protected from ROS-mediated apoptosis. The neuroprotective effects could be attributed to significant amounts of adenosine and herierin III isolated from HESAE.ConclusionsHESAE demonstrated neuroprotective effects against high-dose corticosterone-induced oxidative stress in an in vitro model mimicking depression. HESAE could be a potential dietary supplement to treat depression.

Pyruvate Kinase M2 Coordinates Metabolism Switch between Glycolysis and Glutaminolysis in Cancer Cells.

SummaryCancer cells alter their nutrition metabolism to cope the stressful environment. One important metabolism adjustment is that cancer cells activate glutaminolysis in response to the reduced carbon from glucose entering into the TCA cycle due to inactivation of several enzymes in glycolysis. An important question is how the cancer cells coordinate the changes of glycolysis and glutaminolysis. In this report, we demonstrate that the pyruvate kinase inactive dimer PKM2 facilitates activation of glutaminolysis. Our experiments show that growth stimulations promote PKM2 dimer. The dimer PKM2 plays a role in regulation of glutaminolysis by upregulation of mitochondrial glutaminase I (GLS-1). PKM2 dimer regulates the GLS-1 expression by controlling internal ribosome entry site (IRES)-dependent c-myc translation. Growth stimulations promote PKM2 interacting with c-myc IRES-RNA, thus facilitating c-myc IRES-dependent translation. Our study reveals an important linker that coordinates the metabolism adjustment in cancer cells.

Progress in the Synthesis, Angiogenesis Activity and Mechanism of Chalcone Derivatives

Cancer is a common disease that poses a serious threat to human health. Angiogenesis is essential for the growth and metabolism of tumors, providing oxygen and nutrition for the growth of cells and tissues. However, angiogenesis of tumors depends on the stimulation of growth factors. Vascular Endothelial Growth Factor (VEGFR) is the most unique factor. Therefore, VEGF/VEGFR targeting anticancer drugs are playing an increasingly significant role in clinical trials. In addition, it has been proved that chalcone, the precursor of natural flavonoids, has potential anti-tumor activity, especially anti-angiogenesis activity. This review summarizes the reports about the anti-angiogenesis of chalcone derivatives. Based on the chalcone skeleton, it is divided into substituted chalcones and modified chalcones. The anti-angiogenesis activities of natural or synthetic chalcones, benzene ring modified or connecting bridge modified chalcones are described in this review.