MiR-155 regulates lymphoma cell proliferation and apoptosis through targeting SOCS3/JAK-STAT3 signaling pathway.

Since this article has been suspected of research misconduct and the corresponding authors did not respond to our request to prove originality of data and figures, "MiR-155 regulates lymphoma cell proliferation and apoptosis through targeting SOCS3/JAK-STAT3 signaling pathway, by X.-D. Li, X.-M. Li, J.-W. Gu, X.-C. Sun, published in Eur Rev Med Pharmacol Sci 2017; 21 (22): 5153-5159-DOI: 10.26355/eurrev_201711_13832-PMID: 29228427" has been withdrawn. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/13832.

MiR-155 regulates lymphoma cell proliferation and apoptosis through targeting SOCS3/JAK-STAT3 signaling pathway.

OBJECTIVE: Janus kinase (JAK)- signal transducer and activator of transcription (STAT) signaling pathway participates in regulating cell proliferation, differentiation, and apoptosis, and related to lymphoma. Suppressors of cytokine signaling 3 (SOCS3) is a negative regulator of the JAK-STAT signaling pathway. SOCS3 reduction and miR-155 up-regulation are associated with lymphoma pathogenesis. Bioinformatics analysis showed the complementary binding site between miR-155 and SOCS3. This study aimed to investigate the role of miR-155 in regulating SOCS3/JAK-STAT signaling pathway and affecting diffuse large B cell lymphoma (DLBCL) cell proliferation and apoptosis. PATIENTS AND METHODS: DLBCL tumor sample was collected from the patients in our hospital. Lymphatic tissue derived from reactive lymphoid hyperplasia patients were selected as control. MicroRNA-155 (MiR-155) and SOCS3 expressions were detected. Dual luciferase assay was used to verify the targeted relationship between miR-155 and SOCS3. OCI-LY10 cells were cultured in vitro and divided into five groups, including miR-NC, miR-155 inhibitor, pIRES2-Blank, pIRES2-SOCS3, and miR-155 + pIRES2-SOCS3 groups. SOCS3, p-JAK1, p-JAK2, p-STAT3, and Survivin expressions were tested. Cell apoptosis and proliferation were detected by flow cytometry. RESULTS: MiR-155 expression significantly increased, while SOCS3 level declined in DLBCL tissue compared with control. MiR-155 targeted regulated SOCS3 expression. MiR-155 inhibitor and/or pIRES2-SOCS3 transfection markedly up-regulated SOCS3 expression, reduced p-JAK1, p-JAK2, p-STAT3, and Survivin levels, attenuated cell proliferation, and enhanced cell apoptosis in OCI-LY10 cells. CONCLUSIONS: Down-regulation of miR-155 inhibited DLBCL cell proliferation and facilitated apoptosis through up-regulating SOCS3 expression to suppress JAK-STAT3 signaling pathway.

The PPI network and clusters analysis in glioblastoma.

OBJECTIVE: Glioblastoma is the most aggressive tumor of the brain. To further understand its molecular mechanism, we carried out a systemic bioinformatics study of gene chips downloaded from Gene Expression Omnibus database. MATERIALS AND METHODS: LIMMA package in R language was used to identify the differentially expressed genes (DEGs) between glioblastoma samples and normal controls. RESULTS: Further, we constructed protein-protein interaction networks by mapping the DEGs into PPI data and identified network clusters in these networks. The results revealed that expression of 516 genes, which are mainly involved in phosphate metabolic process and signal transduction, were altered in glioblastoma samples. LYN, CD22 and LCP2 form a densely protein complex in the PPI network. CONCLUSIONS: Our results suggest that LYN, CD22 and LCP2 play important roles in the occurrence and progression of glioblastoma.

Analysis of key genes and modules during the courses of traumatic brain injury with microarray technology.

Gene expression data acquired at different times after traumatic brain injury (TBI) were analyzed to identify differentially expressed genes (DEGs). Interaction network analysis and functional enrichment analysis were performed to extract valuable information, which may benefit diagnosis and treatment of TBI. Microarray data were downloaded from Gene Expression Omnibus and pre-treated with MATLAB. DEGs were screened out with the SAM method. Interaction networks of the DEGs were established, followed by module analysis and functional enrichment analysis to obtain insight into the molecular mechanisms. A total of 39 samples at six time points (30 min, 4, 8, 24 , 72 h, and 21 days) were analyzed and generated 377 DEGs. Eight modules were identified from the networks and network ontology analysis revealed that cell surface receptor-linked signaling pathway, response to wounding and signaling pathway were significantly overrepresented. Altered risk genes and modules in TBI were uncovered through comparing the gene expression data acquired at various time points. These genes or modules could be potential biomarkers for diagnosis and treatment of TBI.

Bioinformatics analysis of gene expression profiles in the rat cerebral cortex following traumatic brain injury.

BACKGROUND: Traumatic brain injury (TBI) is a serious neurodisorder commonly caused by sports related events or violence. It is the leading cause of disability in people under 40. AIM: In order to elucidate the molecular mechanism of the secondary injury after TBI. MATERIALS AND METHODS: In this study, we downloaded gene expression profile on TBI model with sham controls for gene set enrichment analysis and pathway analysis. RESULTS: At a q-value of 5%, 361 genes were up-regulated and 373 were down-regulated in samples obtained at 48 hours after TBI. Functional analyses revealed that steroid biosynthesis, cell cycle, metal ion transport, inflammation and apoptosis were significantly dysregulated during the late period after trauma. In addition, MAPK3 (mitogen-activated protein kinase 3), was identified as the hub node in the protein-protein interaction (PPI) network constructed by the differentially expressed genes (DEGs). CONCLUSIONS: Further elucidation of genes and proteins in our study may reveal their potential as novel therapeutic targets.

The association between lysosomal protein glucocerebrosidase and Parkinson's disease.

BACKGROUND: In recent years, mutations in glucocerebrosidase gene (GBA), which encodes the lysosomal enzyme glucocerebrosidase (GCase) deficient in Gaucher disease (GD), were found to be the most widespread genetic for the development of Parkinson disease. AIM: In this work, we investigated the possibility of a biological linkage between GCase and alpha-synuclein. MATERIALS AND METHODS: siRNA was used to knockdown the GBA, then the related proteins such as alpha-synuclein were detected, additionally, the mutations of GBA were also detected. We also provide evidence that a mouse model of Gaucher disease (GBAD409H/D409H) to detect the gene types of GBA. RESULTS: The results showed functional knockdown (KD) of GBA in neuroblastoma cells culture causes a significant accumulation of alpha-synuclein and alpha-synuclein-mediated neurotoxicity. Furthermore, KD of GBA in rat primary neurons expressing the A53T mutation of alpha-synuclein, decreases cell viability. In addition, we observed that overexpression of several GBA mutants (N370S, L444P, D409H, D409V) significantly raised human alpha-syn levels of vector control. Glucosylceramide (GlcCer), the GCase substrate, influenced formation of purified a-syn by stabilizing soluble oligomeric intermediates. We also provide evidence that a mouse model of Gaucher disease (GBAD409H/D409H) exhibited alpha-syn aggregates in substantia nigra, cortex and hippocampus regions. ELISA analysis showed a significant rise in membrane-associated α-syn and western blot analysis showed that two forms of alpha-syn oligomers were present in brain homogenates from the hippocampus D409H mice. CONCLUSIONS: These studies support the contention that both WT and mutant GBA can cause Parkinson disease-like alpha-synuclein pathology.

Interactions between oxidative stress and inflammation in salt-sensitive hypertension.

The goal of this study was to test the hypothesis that increases in oxidative stress in Dahl S rats on a high-salt diet help to stimulate renal nuclear factor-kappaB (NF-kappaB), renal proinflammatory cytokines, and chemokines, thus contributing to hypertension, renal damage, and dysfunction. We specifically studied whether antioxidant treatment of Dahl S rats on high Na intake would decrease renal inflammation and thus attenuate the hypertensive and adverse renal responses. Sixty-four 7- to 8-wk-old Dahl S or R/Rapp strain rats were maintained for 5 wk on high Na (8%) or high Na + vitamins C (1 g/l in drinking water) and E (5,000 IU/kg in food). Arterial and venous catheters were implanted at day 21. By day 35 in the high-Na S rats, antioxidant treatment significantly increased the renal reduced-to-oxidized glutathione ratio and decreased renal cortical H(2)O(2) and O(2)(*-) release and renal NF-kappaB. Antioxidant treatment with vitamins C and E in high-Na S rats also decreased renal monocytes/macrophages in the glomeruli, cortex, and medulla, decreased tumor necrosis factor-alpha by 39%, and decreased monocyte chemoattractant protein-1 by 38%. Vitamin-treated, high-Na S rats also experienced decreases in arterial pressure, urinary protein excretion, renal tubulointerstitial damage, and glomerular necrosis and increases in glomerular filtration rate and renal plasma flow. In conclusion, antioxidant treatment of high-Na Dahl S rats decreased renal inflammatory cytokines and chemokines, renal immune cells, NF-kappaB, and arterial pressure and improved renal function and damage.

Immune suppression prevents renal damage and dysfunction and reduces arterial pressure in salt-sensitive hypertension.

The goal of this study was to test the hypothesis that renal infiltration of immune cells in Dahl S rats on increased dietary sodium intake contributes to the progression of renal damage, decreases in renal hemodynamics, and development of hypertension. We specifically studied whether anti-immune therapy, using mycophenolate mofetil (MMF), could help prevent increases in renal NF-kappaB activation, renal infiltration of monocytes/macrophages, renal damage, decreases in glomerular filtration rate (GFR) and renal plasma flow, and increases in arterial pressure. Seventy-four 7-to 8-wk-old Dahl S, Rapp strain rats were maintained on an 8% Na, 8% Na + MMF (20 mg.kg(-1).day(-1)), 0.3% Na, or 0.3% Na + MMF diet for 5 wk. Arterial and venous catheters were implanted at day 21. By day 35, renal NF-kappaB in 8% Na rats was 47% higher than in 0.3% Na rats and renal NF-kappaB was 41% lower in 8% Na + MMF rats compared with the 8% Na group. MMF treatment significantly decreased renal monocyte/macrophage infiltration and renal damage and increased GFR and renal plasma flow. In high-NA Dahl S rats mean arterial pressure increased to 182 +/- 5 mmHg, and MMF reduced this arterial pressure to 124 +/- 3 mmHg. In summary, in Dahl S rats on high sodium intake, treatment with MMF decreases renal NF-kappaB and renal monocyte/macrophage infiltration and improves renal function, lessens renal injury, and decreases arterial pressure. This suggests that renal infiltration of immune cells is associated with increased arterial pressure and renal damage and decreasing GFR and renal plasma flow in Dahl salt-sensitive hypertension.

Preparation of bioelectret collagen and its influence on cell culture in vitro.

Collagen extracted from pigskin was corona-charged negatively by a specifically designed device. Different charging voltages, temperatures and times were applied to prepare collagen bioelectret. The decline of the surface potential of the bioelectret under different treatment was then determined. The data showed that the surface potential was markedly varied with the charging conditions. The optimal values of three parameters for charging collagen coatings were defined as follows: voltage, 8 KV; temperature, 40 degrees Celsius; time, 25 min. Treatment of the bioelectret with distillated water, or saline solution (0.9%) or culture medium induced a sharp decrease of the surface potential. In addition, we investigated the effects of the charged collagen on cell growth and intracellular calcium level of three types of cultured mammalian cell lines, including Chinese hamster ovary CHO cells, human cervix uteri tumor HeLa cells and human promyelocytic HL-60 cells. Cell growth and the intracellular calcium level were determined by MTT reagent-based assay and a fluorescent probe Fura-2, respectively. The results showed that negatively charged collagen stimulated the growth of CHO or HL-60 cell line but inhibited the growth of HeLa cell line. Furthermore, after attaching to the charged collagen, the intracellular calcium level of CHO cells increased, while that of HeLa cells decreased. Thus we proposed for the first time that collagen bioelectret could differentially modulate the growth of different cells, by an unknown mechanism that probably involves a role of intracellular calcium.

Moderate levels of ethanol induce expression of vascular endothelial growth factor and stimulate angiogenesis.

Alcohol abuse has a negative impact on human health; however, epidemiological studies show that moderate consumption of ethanol (EtOH) reduces the risk of coronary heart disease, sudden cardiac death, and ischemic stroke. The mechanisms for these reductions in cardiovascular disease are not well established. Using cultured coronary artery vascular smooth muscle cells, we found that moderate levels of EtOH (10 and 20 mM) caused dose-related increases in both vascular endothelial growth factor (VEGF) mRNA (Northern blot) expression (1.9- and 2.6-fold) and VEGF protein (ELISA) expression (19 and 68%) compared with control (P < 0.05). EtOH at 0.25 g. kg(-1). day(-1) (7 days) increased VEGF mRNA expression by 1.48-fold over control, and increased vessel length density from 3.9 +/- 0.7 (control) to 6.0 +/- 0.3 mm/mm(2) (P < 0.05) in chick chorioallantoic membrane (CAM). We conclude that moderate levels of ethanol can induce VEGF expression and stimulate angiogenesis in chick CAM. Therefore, the results provide a theoretical basis for speculating that the cardiovascular-protective effects of moderate alcohol consumption may be partly mediated through VEGF-induced angiogenesis.

[The influence of acetylcholine on the proliferation of cultured human pituitary tumour cells].

AIM: To study the influence of acetylcholine on the proliferation, DNA synthesis and cell cycle of cultured human pituitary tumour cells. METHODS: MTT method, 3H-TdR incorporation and cell cycle analysis were used to examine the changes of proliferation and DNA synthesis of human pituitary tumour cells. RESULTS: Ach at 10(-7) mol/L - 10(-5) mol/L could decrease the 3H-TdR incorporation and the MTT A value in a dose dependent manner (P < 0.01), and the ratio of G1 phase of pituitary tumour cells increased markedly (P < 0.01). The effect of acetylcholine on the proliferation of cultured human pituitary tumour cells could be inhibited by atropine. CONCLUSION: Ach inhibited the proliferation, DNA synthesis and cell cycle of cultured human pituitary tumour cells, and the inhibitory effect was mediated by acetylcholine receptor.

[Experimental studies of genistein on the proliferation and apoptosis of human prolactinoma cells cultured in vitro].

AIM: To study the influence of genistein (GST) on the proliferation and apoptosis of cultured human prolactinoma cells. METHODS: MTT method and 3H-TdR incorporation and cell cycle analysis were used to examine the changes of proliferation and DNA synthesis of human prolactinoma cells under influence of GST and beta-estradiol (E2). Tdt-mediated dUTP nick end labeling (TUNEL) were employed to observe the effect of GST and E2 on the apoptosis of human prolactinoma cells. RESULTS: In a dose dependent manner, GST of different concentration could significantly inhibit the proliferation of human prolactinoma cells cultured in vitro. GST(10(-5) mol/L) could increase the proportion of cells in G1 phase from 55.3% up to 90.3%. E2 of different concentration could dose-dependently increase the proliferation of human prolactinoma cells. E (10(-5) mol/L) could increase the proportion of cells in G2 phase from 15.6% up to 41.8%. However, a lower suppressive proliferation of cultured human prolactinoma cells was observed with GST and E2 together. GST, not E2, could significantly induce the apoptosis of human prolactinoma cells cultured in vitro. CONCLUSION: GST inhibits the proliferation, DNA synthesis and cell cycle of cultured human pituitary cells, and induces its apoptosis. E2 decreases partly the effect of GST on the suppression of proliferation, not apoptotic induction, of human prolactinoma cells cultured in vitro.

Inhibition of adenosine kinase induces expression of VEGF mRNA and protein in myocardial myoblasts.

We tested whether increased endogenous adenosine produced by the adenosine kinase inhibitor GP-515 (Metabasis Therapeutics) can induce vascular endothelial growth factor (VEGF) expression in cultured rat myocardial myoblasts (RMMs). RMMs were cultured for 18 h in the absence (control) and presence of GP-515, adenosine (Ado), adenosine deaminase (ADA), or GP-515 + ADA. GP-515 (0.2-200 microM) caused a dose-related increase in VEGF protein expression (1.99-2.84 ng/mg total cell protein); control VEGF was 1.84 +/- 0.05 ng/mg. GP-515 at 2 and 20 microM also increased VEGF mRNA by 1.67- and 1. 82-fold, respectively. ADA (10 U/ml) decreased baseline VEGF protein levels by 60% and completely blocked GP-515 induction of VEGF. Ado (20 microM) and GP-515 (20 microM) caused a 59 and 39% increase in VEGF protein expression and a 98 and 33% increase in human umbilical vein endothelial cell proliferation, respectively, after 24 h of exposure. GP-515 (20 microM) had no effect on VEGF protein expression during severe hypoxia (1% O(2)) but increased VEGF by an additional 27% during mild hypoxia (10% O(2)). These results indicate that raising endogenous levels of Ado through inhibition of adenosine kinase can increase the expression of VEGF and stimulate endothelial cell proliferation during normoxic and hypoxic conditions.

Adenosine upregulates VEGF expression in cultured myocardial vascular smooth muscle cells.

We tested whether adenosine has differential effects on vascular endothelial growth factor (VEGF) expression under normoxic and hypoxic conditions, and whether A(1) or A(2) receptors (A(1)R; A(2)R) mediate these effects. Myocardial vascular smooth muscle cells (MVSMCs) from dog coronary artery were exposed to hypoxia (1% O(2)) or normoxia (20% O(2)) in the absence and presence of adenosine agonists or antagonists for 18 h. VEGF protein levels were measured in media with ELISA. VEGF mRNA expression was determined with Northern blot analysis. Under normoxic conditions, the adenosine A(1)R agonists, N(6)-cyclopentyladenosine and R(-)-N(6)-(2-phenylisopropyl)adenosine did not increase VEGF protein levels at A(1)R stimulatory concentrations. However, adenosine (5 microM) and the adenosine A(2)R agonist N(6)-[2-(3, 5-dimethoxyphenyl)-2-(2-methylphenyl)]ethyl adenosine (DPMA; 100 nM) increased VEGF protein levels by 51 and 132% and increased VEGF mRNA expression by 44 and 90%, respectively, in cultured MVSMCs under normoxic conditions. Hypoxia caused an approximately fourfold increase in VEGF protein and mRNA expression, which could not be augmented with exogenous adenosine, A(2)R agonist (DPMA), or A(1)R agonist [1,3-diethyl-8-phenylxanthine (DPX)]. The A(2)R antagonist 8-(3-chlorostyryl)-caffeine completely blocked adenosine-induced VEGF protein and mRNA expression and decreased baseline VEGF protein levels by up to approximately 60% under normoxic conditions but only by approximately 25% under hypoxic conditions. The A(1)R antagonist DPX had no effect. These results are consistent with the hypothesis that 1) adenosine increases VEGF protein and mRNA expression by way of A(2)R. 2) Adenosine plays a major role as an autocrine factor regulating VEGF expression during normoxic conditions but has a relatively minor role during hypoxic conditions. 3) Endogenous adenosine can account for the majority of basal VEGF secretion by MVSMCs under normoxic conditions and could therefore be a maintenance factor for the vasculature.

Sodium induces hypertrophy of cultured myocardial myoblasts and vascular smooth muscle cells.

The mechanisms of sodium-induced myocardial hypertrophy and vascular hypertrophy are poorly understood. We tested the hypothesis that a high sodium concentration can directly induce cellular hypertrophy. Neonatal rat myocardial myoblasts (MMbs) and vascular smooth muscle cells (VSMCs) were cultured in a 50:50 mixture of DMEM and M199 supplemented with 10% fetal bovine serum. When the monolayers reached approximately 80% confluence, normal sodium medium (146 mmol/L) was replaced with high sodium media (152 mmol/L, 160 mmol/L, and 182 mmol/L) for up to 5 days. Increasing sodium from a baseline concentration of 146 mmol/L to the higher concentrations for 5 days caused dose-related increases in cell mean diameter, cell volume, and cellular protein content in both MMbs and VSMCs. Increasing the sodium concentration by only 4% (from 146 mmol/L to 152 mmol/L) caused the following respective changes in MMbs and VSMCs: 8.5% and 8.7% increase in cell mean diameter, 27.6% and 27.0% increase in cell volume, and 55.7% and 46.7% increase in cellular protein content. The rate of protein synthesis, expressed as [3H]leucine incorporation, increased by 87% and 99% in MMbs after exposure to 152 mmol/L and 160 mmol/L sodium, respectively, compared with the 146-mmol/L sodium control group. Exposure of MMbs to medium with a sodium concentration of 10% above normal, ie, 160 mmol/L, caused a significant decrease (range, 26% to 44%) in the rate of protein degradation at multiple time points over a 48-hour period compared with normal sodium control cells. The increase in cellular protein content caused by 160 mmol/L sodium returned to normal within 3 days after MMbs were returned to a normal sodium medium. These findings support the hypothesis that sodium has a direct effect to induce cellular hypertrophy and may therefore be an important determinant in causing myocardial and/or vascular hypertrophy in subjects with increased sodium concentration in the extracellular fluid.

Hypoxia-induced expression of VEGF is reversible in myocardial vascular smooth muscle cells.

We determined whether hypoxia-induced expression of vascular endothelial growth factor (VEGF) can be reversed by a normoxic environment. Dog myocardial vascular smooth muscle cells (MVSMCs) were exposed to hypoxia (1% O2) for 24 h and then returned to normoxia (20% O2). VEGF protein levels increased by more than fivefold after 24 h of hypoxia and returned to baseline within 24 h of the return of the cells to normoxia. Northern blot analysis showed that hypoxia caused a 5.5-fold increase in VEGF mRNA, and, again, the expression was reversed after reinstitution of normoxia. Additional measurements showed that basic fibroblast growth factor and platelet-derived growth factor protein levels were not induced by hypoxia and that hypoxia caused a fourfold decrease in transforming growth factor-beta 1 protein levels. Hypoxia conditioned media from MVSMCs caused human umbilical vein endothelial cells to increase [3H]thymidine incorporation by twofold, an effect that was blocked in a dose-dependent manner by anti-human VEGF antibody. The hypoxia conditioned media had no effect on MVSMC proliferation. These findings suggest that VEGF expression can be bidirectionally controlled by tissue oxygenation, and thus support the hypothesis that VEGF is a physiological regulator of angiogenesis.

Basic fibroblast growth factor as a biochemical marker of exercise-induced ischemia.

BACKGROUND: Basic fibroblast growth factor (bFGF) is a mitogenic polypeptide that demonstrates enhanced expression and promotes angiogenesis in animal models of myocardial ischemia and infarction. Elevated levels of bFGF are present in the urine of humans with metastatic tumors, but its expression in human myocardial ischemia is unknown. Thus, we sought to determine whether urine levels of bFGF are altered by exercise-induced ischemia in humans. METHODS AND RESULTS: Eighty-six patients underwent exercise thallium studies for evaluation of anginal symptoms. Urine levels of bFGF (corrected for urine creatinine) were determined by ELISA immediately before and between 2 and 4 hours after exercise. The change in urine bFGF level was compared between 43 patients with and 43 patients without exercise-induced ischemia. Patients with ischemia had an increase in urine bFGF compared with nonischemic patients (1052 +/- 245 versus -278 +/- 130 pg/g creatinine, P < .0001). Exercise, demographic, and clinical variables were assessed and analyzed for possible effect on bFGF response to exercise. By univariate analysis, a history of hypertension was negatively associated with a change in bFGF level (P < .05). No other variables were associated. By multivariate analysis, only bFGF response (P < .001) and age (P < .05) were independently related to exercise-induced ischemia. CONCLUSIONS: Significantly increased levels of bFGF are detected in the urine within hours of exercise-induced ischemia. Further studies are warranted to determine whether bFGF might serve as a useful circulating marker of myocardial ischemia in humans.