Effect of negative regulation of nucleophosmin 1 by long non-coding RNA growth arrest specific transcript 5 on cisplatin resistance of gastric cancer cells / 中国生物制品学杂志

@#Objective To investigate the effects of long non-coding RNA(LncRNA) growth arrest specific transcript 5(GAS5) negatively regulating nucleophosmin 1(NPM1) on cisplatin(DDP) resistance of gastric cancer cells.Methods The normal human gastric mucosa cell line GES-1 and human gastric cancer cell lines BG3-823,MGC-803 and AGS were selected as the research objects,of which the level of LncRNA GAS5 in each cell was measured by qRT-PCR.The drug resistance of AGS cells to DDP(AGS/DDP) was induced,and the experiment was divided into control group,empty plasmid group(BC group),GAS5 nonsense interference group(pLJM-GAS5 NC group) and GAS5 overexpression group(pLJM-GAS5 group).MTT method was used to determine the effect of DDP on the proliferation of AGS and AGS/DDP cells;and the levels of NPM1,multidrug resistance 1(MDR1),excision repair cross complementation group 1(ERCC1),multidrug resistance-associated protein 1(MRP1) and N-cadherin in AGS and AGS/DDP cells were measured by Western blot.Results Compared with the normal gastric mucosa GES-1 cells,the level of LncRNA GAS5 in BG3-823 and AGS cells decreased significantly,and among them,the level of LncRNA GAS5 in AGS cells was the lowest,so AGS cells were used for the follow-up experiments.Compared with the control group,the level of LncRNA GAS5 in AGS cells of BC group and pLJM-GAS5 NC group decreased significantly,while the levels of NPM1,MDRl,ERCC1,MRP1 and N-cadherin increased significantly;compared with BC group and pLJM-GAS5 NC group,the level of LncRNA GAS5 in AGS/DDP cells of pLJM-GAS5 group increased significantly,while the levels of NPM1,MDR1,ERCC1,MRP1 and N-cadherin decreased significantly;after treatment with DDP of the same concentration(except 0 μmol/L),compared with the control group,the inhibition rate of AGS/DDP cell proliferation in BC group and pLJM-GAS5 NC group decreased significantly;compared with BC group and pLJM-GAS5 NC group,the inhibition rate of AGS/DDP cell proliferation in pLJM-GAS5group was significantly higher.The semi inhibitory concentration(IC_(50)) of DDP on AGS/DDP cells in pLJM-GAS5 group for 48 h was(65.38±5.04) μmol/L,which was significantly lower than(120.74±4.17) μmol/L and(120.24±4.29) μmol/L in BC group and pLJM-GAS5 NC group.Conclusion Up-regulating the level of LncRNA GAS5 in AGS/DDP cells can reverse the drug resistance of AGS/DDP cells,which may be related to the down-regulation of NPM1expression

Research advances in the association of the tyrosine kinase receptor Axl and its ligand Gas6 with hepatocellular carcinoma / 临床肝胆病杂志

As one of the tyrosine kinase receptors, Axl is an important downstream regulator of epithelial-mesenchymal transition (EMT) and can bind to its ligand Gas6 protein. By activating the downstream signal transduction pathways, Axl is closely associated with vascular invasion, tumor metastasis, recurrence, and low survival rate of hepatocellular carcinoma. Current studies on the Gas6/Axl signaling pathway have confirmed that Axl inhibitors play an important role in the treatment of liver cancer and the Gas6/Axl signaling pathway may be a potential therapeutic target for liver cancer. This article mainly introduces the association of Axl and its ligand Gas6 with the development and progression of liver cancer and their application in the diagnosis and treatment of liver cancer, in order to provide new ideas for the early diagnosis of liver cancer and clinical research on anti-cancer treatment.

Screening of Differential Genes for Endometriosis and Expression Validation of the Potential Target GAS6 / 中国生物化学与分子生物学报

Growth arrest specific protein 6 (GAS6) plays an important role in the occurrence and development of tumors, and its signal transduction is involved in cell proliferation, adhesion and migration, but its related functions and molecular mechanisms in endometriosis (EMs) are still unclear. In this study, we searched and downloaded the transcriptome datasets of EMs from GEO database and performed GEO online analysis, and then screened out the differentially expressed genes and performed cluster analysis based on GO and KEGG pathway. The mRNA levels of the differentially expressed genes shared by more than three datasets were verified by qRT-PCR in the endometrium of ten women with no endometriosis and no clear disease and the ectopic endometrium of 11 patients with ovarian chocolate cysts. Immunohistochemistry and qRT-PCR were used to verify the expression of GAS6 and epithelial mesenchymal transition (EMT) marker genes, and immunofluorescence was used to co-label GAS6 and E-cadherin in endometriosis clinical samples. In this study, a total of 47 differentially expressed genes were screened out of the four transcriptome datasets, which were mainly enriched in processes such as cell migration and related signal pathways such as MAPK, PI3K-AKT, and tight junction. The mRNA levels of the nine differentially expressed genes shared by more than three datasets in endometriosis patients were consistent with the results of bioinformatics analysis. GAS6 expression levels in ectopic endometrium of EMs patients are higher than the control group (P < 0. 05), and EMs patients have the characteristics of EMT in the ectopic endometrial tissue, that is, the expression of E-cadherin is down-regulated (P < 0. 05) and the expression of vimentin is up-regulated (P < 0. 01). The expression of E-cadherin in the ectopic endometrial glandular epithelial cells of EMs patients is low while the expression of GAS6 is up-regulated, suggesting that GAS6 may mediate the EMT process in endometriosis. In conclusion, this study reveals that GAS6 is highly expressed in endometriosis patients and may mediate the EMT process to participate in the occurrence and development of endometriosis, providing a potential target for clinical treatment of endometriosis.

Protective effect of glutamine on hyperoxic lung injury of neonatal rats through endoplasmic reticulum stress pathway / 吉林大学学报(医学版)

Objective: To explore the protective effect of glutamine (GLN) on the hyperoxia-induced lung injury of the neonatal rats through endoplasmic reticulum stress (ERS) pathway, and to elucidate its mechanisms. Methods: A total of 90 Wistar rats were randomly divided into control group (FiO2 =21%), hyperoxia group (FiO2 85%), and hyperoxia+GLN group (Fi2 85%, the concentration of intraperitoneal injection of GLN was 0. 75 g · kg-1 · d-1); there were 30 rats in each group The body weights and water contents in the lung tissue of the neonatal rats were measured on the 3rd, 7th and 14th days of the experiment. HE staining was used to determine the morphology of lung tissue of the rats. The superoxide dismutase (SOD) activity in lung tissue of the rats was detected by nitro blue tetrazolium chloride (NBT), and the malondialdehyde (MDA) level was determined by thiobarbital acid (TBA). The expression levels of Caspase-12, GADD153, GRP78, Bel-2, and Bax in lung tissue of the rats were detected by Western blotting method. Results: Compared with control group at the same time, the body weights of the neonatal rats in hyperoxia group on the 3rd, 7th and 14th days were significantly decreased (P<0. 05), the water contents in lung tissue of the neonatal rats were increased (P<0. 05), the SOD activities were significantly decreased (P<0. 05), the levels of MDA in the lung tissue of the neonatal rats were increased (P<0. 05), the expressions levels of Caspase-12, GADD153, GRP78 and Bax proteins were significantly increased (P<0. 05), and the expression levels of Bcl-2 protein and the Bcl-2/Bax ratios were significantly decreased (P<0. 05). Compared with hyperoxia group at the same time, the body weights of the neonatal rats in hyperoxia + GLN group on the 3rd, 7th and 14th days were significantly increased (P<0. 05), the water contents in lung tissue of the neonatal rats were decreased (P<0. 05), the SOD activities were significantly increased (P< 0. 05), the levels of MDA in lung tissue of the neonatal rats were decreased (P<0. 05), the expression levels of Caspase-12, GADD153, GRP78 and Bax proteins were significantly decreased (P<0. 05), the expression levels of Bcl-2 protein and the Bcl-2/Bax ratios were increased (P<0. 05). The pathological sections of lung tissue of the rats in control group showed that lung tissue structure was regular, no alveolar edema was found, the alveolar size and alveolar septum were approximately the same, and no inflammatory cell infiltration was found; the histopathological sections of lung tissue of the rats in hyperoxia group showed swelling of brochial and alveolar epithelial cells, enlargement of alveolar lumen, edema of interstitial cells, inflammatory cell infiltration and fibrous exudation; the degrees of alveolar damage, the inflammatory exudation and the proliferation of fibrons tissue in hyperoxia+GLN group were alleviated which was between hyperoxia group and control group. Conclusion: GLN can alleviate the hyperoxia-induced lung tissue edema and inflammatory response of the neonatal rats, and one of mechanisms is that GLN can down-regulate the expression levels of Caspase-12, GADD153, GRP78 and Bax proteins and up-regulate the expression level of Bcl-2 protein through ERS pathway to protect hypoxic lung injury.

Role of growth arrest-specific protein 6 in migration and osteogenic differentiation of human periodontal ligament cells / 北京大学学报(医学版)

OBJECTIVE@#To investigate the role of growth arrest-specific protein 6 (Gas6) in the process of the migration and osteogenic differentiation of human periodontal ligament cells (hPDLCs).@*METHODS@#After different concentrations of recombinant human Gas6 (rhGas6) were added to hPDLCs, cell prolife-ration experiment (CCK-8) was taken to observe the effect of rhGas6 on hPDLCs cell proliferation. Scratch test and cell migration test (Transwell) were taken to analyze the migratory ability of hPDLCs in different concentrations of rhGas6 groups. After osteogenic induction, real-time quantitative polymerase chain reaction (real-time PCR) was taken to detect the expression of the Runt-related transcription factor 2 (Runx2) and alkaline phosphatase (ALP). ALP staining was used to detect the amount of mineralized nodules.@*RESULTS@#After adding different concentrations of rhGas6, there were no statistically significant differences in hPDLCs cell proliferation among the experimental groups and the control group at 24, 48 and 72 hours (P>0.05). After 24 h of scratch, the healing area in the 800 μg/L of the rhGas6 group was greater than that in the control group, but without statistically significant difference (31.06%±13.70% vs. 21.79%±9.51%, P>0.05). In the migration test, after 24 h, the number of hPDLCs cells which penetrated through the membrane in the 800 μg/L rhGas6 group was significantly higher than that in the control group (P < 0.01). After rhGas6 was added and osteogenic induction, Runx2 and ALP gene expressions of hPDLCs in the 800 μg/L group were significantly higher than those in the control group (1.60±0.30 vs. 0.91±0.10, 2.81±0.61 vs. 0.86±0.12, P < 0.01). After Gas6 was knocked down, the ALP expression of hPDLCs was significantly lower than that of the control group (0.39±0.07 vs. 0.92±0.14, P < 0.01). There was no significant change in Runx2 expression (P>0.05). After 7 days of osteogenic induction, the mineralized nodules formed in the Gas6 knockdown group were significantly less than those in control group (0.25±0.04 vs. 1.00±0.11, P < 0.001). After 14 days of induction, the staining degree of the Gas6 knockdown group was lower than that of the control group, but there was no significant difference (0.86±0.04 vs. 1.00±0.16, P>0.05).@*CONCLUSION@#After downregulation of Gas6 gene, mineralized nodule formation was reduced and ALP gene expressions were decreased in the early stage of osteogenic induction (7 days). After addition of rhGas6, Runx2 and ALP gene expressions were increased and the number of cell migration was increased, suggesting that Gas6 might play a promoting role in the migration and osteogenic differentiation of human periodontal ligament cells.

Vascular calcification and growth arrest specific protein 6 levels in chronic renal failure / Calcificación vascular y detención de crecimiento de proteína específica 6 niveles en insuficiencia renal crónica

INTRODUCTION: Cardiovascular disease is the main cause of mortality and morbidity in chronic renal failure. It's known that vascular calcification (VC) and carotid intima media thickness (CIMT) are strongly associated with cardiovascular diseases. Growth arrest specific protein 6 (Gas6) is a vitamin K-dependent protein and regulates various processes such as proliferation, cell survival, migration and inflammation. Gas6 is known to protect endothelial cells and vascular smooth muscle cells against apoptosis by inhibiting Bcl-2 induced Caspase 3 activation. The relationship between Gas6 and cardiovascular diseases has been demonstrated in many mouse models and cell cultures. However, there are conflicting reports whether Gas6 levels are increasing or decreasing in human studies of diabetic and/or chronic renal failure. In present study the aim was to examine plasma Gas6 levels and its relation with CIMT and coronary artery calcification score (CACS) in chronic kidney disease (CKD) patients. METHODS: Total of 137 patients of which 32 chronic hemodialysis and 105 predialysis patients as well as 73 healthy controls were enrolled in the study. Human Gas6 levels in serum samples were studied by ELISA method. CIMT was measured by ultrasonography. CACS was measured by multislice computed tomography. RESULTS: The mean age was 54.37±16.61 years in dialysis group, 55.20±14.80 years in predialysis group and 53.26±9.04 years in control group. Serum creatinine was 0.78±0.16 mg/dl in the control group and 1.96±1.64 mg/dl in the predialysis group and 5.94±1.55 mg/dl in the dialysis group. 24 hours urine protein levels were significally higher in the dialysis group than the predialysis and the control group. CIMT values were similar in predialysis and dialysis groups. These values were significantly higher than the control group. Although CACS was higher in dialysis group than predialysis and control group, the results were not statistically significant since the distribution range was very wide. Gas6 was 98.84±53.32 ng/mL in the control group and statistically higher than the dialysis (63.85±38.92 ng/mL) and the predialysis groups (54.96±38.49 ng/mL) (p=0.001). Gas6 levels were lower in diabetic patients than non-diabetics (53.69±35.26 ng/mL, 69.26±47.50 ng/mL, p=0.023, respectively). Negative correlation was detected between Gas6 and age, BMI, CACS, carotid IMT and proteinuria. In the logistic regression analysis, Gas6 remained significantly associated with BMI, CIMT and proteinuria. CONCLUSION: In our study, a negative correlation of Gas6 with BMI, CACS, CIMT and proteinuria and lower Gas6 levels in diabetic patients support that decreased Gas6 levels in chronic renal failure may have a role in vascular calcification through altered glucose tolerance, chronic inflammation, endothelial dysfunction and increased apoptosis. Our study has an importance because it is the first study showing a relation between Gas6 and proteinuria, CACS and carotid IMT in patients with chronic renal failure

INTRODUCCIÓN: La enfermedad cardiovascular es la principal causa de mortalidad y morbilidad en la insuficiencia renal crónica. Se sabe que la calcificación vascular (CV) y el grosor de la íntima-media de la carótida (CIMT, por sus siglas en inglés) están vinculados de forma muy estrecha con enfermedades cardiovasculares. La proteína específica del gen 6 de la detención de crecimiento (Gas6) es una proteína dependiente de la vitamina K y regula diversos procesos, como la proliferación, la supervivencia celular, la migración y la inflamación. La proteína Gas6 es conocida por proteger las células endoteliales y las células musculares lisas vasculares contra la apoptosis mediante la inhibición de la activación de la caspasa-3 inducida por la proteína Bcl-2. Se ha demostrado la relación entre la Gas6 y las enfermedades cardiovasculares en muchos modelos de ratones y cultivos celulares. Sin embargo, existen informes contradictorios acerca de si los niveles de Gas6 aumentan o disminuyen en estudios de humanos con insuficiencia renal crónica y/o diabética. En este estudio, el objetivo fue examinar los niveles plasmáticos de Gas6 y su relación con el CIMT y la puntuación de calcificación de las arterias coronarias (CACS, por sus siglas en inglés) en pacientes con enfermedad renal crónica (ERC). MATERIAL Y MÉTODOS: Un total de 137 pacientes fueron incluidos en el estudio, de los cuales 32 estaban en hemodiálisis crónica, 105 en prediálisis, y 73 pacientes representaban controles sanos. Se esudiaron los niveles de Gas6 en muestras de suero mediante el método ELISA. El CIMT se midió por medio de ecografía. La CACS se midió mediante tomografía computarizada multicorte. RESULTADOS: La edad media fue de 54,37 ± 16,61 años en el grupo de diálisis; 55,20 ± 14,80 años en el grupo de prediálisis, y 53,26 ± 9,04 años en el grupo de control. La creatinina sérica fue de 0,78 ± 0,16 mg/dl en el grupo de control; 1,96 ± 1,64 mg/dl en el de prediálisis, y 5,94 ± 1,55 mg/dl en el de diálisis. Las concentraciones de proteína en orina de 24 horas fueron significativamente más altas en el grupo de diálisis que en los de prediálisis y control. Los valores del CIMT fueron similares en los grupos de prediálisis y de diálisis. Estos valores fueron considerablemnete más altos que en el grupo de control. Aunque la CACS fue más alta en el grupo de diálisis que en los otros dos, los resultados no fueron estadísticamente significativos, ya que el rango de distribución fue muy amplio. La proteína Gas6 fue de 98,84 ± 53,32 ng/ml en el grupo de control y estadísticamente más alta que en los grupos de diálisis (63,85 ± 38,92 ng/ml) y de prediálisis (54,96 ± 38,49 ng/ml) (p = 0,001). Los niveles de Gas6 fueron más bajos en los pacientes diabéticos que en los no diabéticos (53,69 ± 35,26 ng/ml; 69,26 ± 47,50 ng/ml, [p = 0,023], respectivamente). Se detectó una correlación negativa entre la proteína Gas6 y la edad, el IMC, la CACS, el CIMT y la proteinuria. En el análisis de regresión logística, la Gas6 se mantuvo estrechamente relacionada con el IMC, el CIMT y la proteinuria. CONCLUSIÓN: En nuestro estudio, la correlación negativa de Gas6 con IMC, CACS, CIMT y proteinuria, y los niveles más bajos de Gas6 en pacientes diabéticos sustentan la idea de que la disminución de los niveles de Gas6 en la insuficiencia renal crónica puede jugar un papel en la calcificación vascular a través de la tolerancia alterada a la glucosa, la inflamación crónica, la disfunción endotelial y el aumento de la apoptosis. La importancia de nuestro estudio radica en que es el primero que muestra una relación entre la Gas6 y la proteinuria, la CACS y el CIMT en pacientes con insuficiencia renal crónica

Effect of electroacupuncture on expressions of Gas7 and NGF in arcular nucleus of rats with focal cerebral ischemia / 中国针灸

OBJECTIVE@#To observe the effect of electroacupuncture (EA) on the expressions of growth arrest-specific protein 7 (Gas7) and nerve growth factor (NGF) in arcuate nucleus (ARC) of rats with focal cerebral ischemia and explore the potential action mechanism of EA in treatment of focal cerebral ischemia.@*METHODS@#A total of 50 SD rats were randomized into 4 groups, named a normal group ( =12), a sham-operation group ( =12), a model group ( =14) and an EA group ( =12). In the model group and the EA group, the thread embolization method was adopted to duplicate the model of the right middle cerebral arterial embolism. In the sham-operation group, the skin of the neck was opened and sutured without any other intervention. In the EA group, EA was applied to "Baihui" (CV 20) and "Zusanli" (ST 36) on the left side, once a day, 30 min each time, consecutively for 21 days, while there was no any intervention in the normal group, the sham-operation group and the model group. Using the immunohistochemistry (IHC) method and Western blot method, the expressions of Gas7 and NFG of ARC on the ischemic side were determined. Using Nissle staining, the morphological changes in ARC neurons were observed.@*RESULTS@#The results of Nissle staining showed that there was no significant change in the morphology of ARC neurons in the normal group and the sham-operation group. In the model group, the volume of neuron cells was atrophied obviously and the cells were arranged irregularly. In the EA group, the morphology of ARC neuron was similar to the normal group. The results of IHC and Western blot indicated that the expressions of immunoreactive neurons and protein of Gas7 and NGF in ARC of the rats in the model group were increased obviously as compared with the normal group and the sham-operation group and the expressions in the EA group were further enhanced as compared with the model group (all <0.05).@*CONCLUSION@#Gas7 and NGF may be participated in the compensatory process of partial protection of the body in the patients with focal cerebral ischemia. EA up-regulates the expressions of Gas7 and NGF in ARC, which may be one of the neuroprotective mechanisms of EA in treatment of cerebral ischemia.

Effects of expression of growth arrest and DNA damage inducible gene 45α interfered by small interference RNA on the invasion and migration of a cutaneous squamous cell carcinoma cell line A431 / 中华皮肤科杂志

Objective To evaluate effects of targeted silencing of growth arrest and DNA damage inducible gene 45α (Gadd45α) by small interference RNA (siRNA) on the invasion and migration of a cutaneous squamous cell carcinoma cell line A431.Methods Real-time fluorescence-based quantitative PCR and Western blot analysis were performed to detect the mRNA and protein expression of A431 and Colon16 cells respectively,and then A431 cells with highly expressed Gadd45α served as a research object.Cultured A431 cells were divided into 3 groups:experimental group transfected with Gadd45α-siRNA-1,negative control group transfected with negative control siRNA,and blank control group receiving no treatment.After the RNA interference,the mRNA and protein expression of Gadd45α in the above 3 groups were measured by real-time fluorescence-based quantitative PCR and Western blot analysis,respectively,and the effect of the RNA interference on the invasion and migration abilities of A431 cells was evaluated by Transwell and wound scratch assays.Results Gadd45α mRNA and protein were highly expressed in the A431 cells.After the RNA interference,the experimental group showed markedly lower mRNA and protein expressions of Gadd45 in the A431 cells (0.286 ± 0.013,0.33 ± 0.007,respectively) compared with the negative control group (1.028 ± 0.183,0.87 ± 0.002,respectively)and blank control group (1.001 ± 0.057,0.86 ± 0.004,respectively),and there were significant differences in the mRNA and protein expressions of Gadd45 among the 3 groups (F =5 893.857,2 763.000,both P ＜ 0.001).The number of A431 cells crossing the polycarbonate membrane was higher in the experimental group (66.33 ± 3.79) than in the negative control group (26.00 ± 4.36) and the blank control group (28.33 ± 4.16),and there was a significant difference among the 3 groups (F =20.084,P =0.002).Moreover,the wound scratch assay showed that the number of migrating A431 cells per high-power field was significantly higher in the experimental group than in the negative control group and the blank control group (315.33 ± 6.66,154.67 ± 2.08,130.67 ± 3.51 respectively;F =1 676.255,P ＜ 0.001).Conclusion Gadd45α is highly expressed in the cutaneous squamous cell carcinoma cell line A431,and targeted silencing of Gadd45α gene can enhance the in vitro invasion and migration abilities of A431 cells.

Effects of miR-130a on viability and apoptosis of rat basilar arterial smooth muscle cells / 中国病理生理杂志

AIM:To investigate the regulatory effects of microRNA (miR)-130a on the biological characteris-tics of rat basilar arterial smooth muscle cells (BASMCs) and its underlying mechanisms. METHODS:The expression of miR-130a in rat BASMCs was measured by real-time PCR. After knockdown of miR-130a with inhibitor in the BASMCs, the cell viability, cell cycle distribution and apoptosis were detected by CCK-8 assay and flow cytometry. The expression of cell cycle- and apoptosis-related molecules, such as cyclin D1, cyclin-dependent kinase 2 (CDK2), p21, Bcl-2 and cleaved caspase-3/caspase-3 at protein levels was determined by Western blot. The growth arrest-specific homeobox protein ( Gax) expression at mRNA and protein levels was determined by real-time PCR and Western blot. RESULTS:Angioten-sionⅡ (AngⅡ) up-regulated the expression of miR-130a and down-regulated the expression of Gax (P<0. 05). Transfec-tion with miR-130a inhibitor partly reversed the increase in AngⅡ-induced cell viability and promoted the Gax expression. Furthermore, the early cell apoptotic rate was significantly increased after down-regulation of miR-130a (P<0.05), and the protein levels of cyclin D1, CDK2, Bcl-2 and p-Rb were significantly decreased, accompanied with the up-regulation of p21 and cleaved caspase-3 (P<0.05). CONCLUSION:Down-regulation of miR-130a restrains the viability and pro-motes the apoptosis of BASMCs by promoting Gax expression and regulating cell cycle- and apoptosis-related molecules, suggesting that miR-130a may be a potential therapeutic target of brain vascular remodeling during hypertension.

Down-regulated expression and antioncogenetic role of GADD45g in acute myeloid leukemia / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To quantify the expression of growth arrest and DNA damage inducible protein 45g (GADD45g) gene in the bone marrow samples of patients withAML (acute myeloid leukemia) and inAML cell lines, as well as to study the correlation between the GADD45g expression and prognostic outcome in patients withAML and investigate the role of GADD45g over-expression in proliferation, apoptosis, senescence, differentiation, cell cycle arrest, and drug sensitivity in AML cell lines. Methods: In the study, a total of 27 cases of bone marrow specimens were selected from patients initially diagnosed as AML in Hospital of Blood Diseases affiliated to Chinese Academy of Medical Sciences from January 2013 to December 2016. mRNA and protein expression levels of GADD45g in BMMNCs (Bone marrow mononuclear cells) from patients with AML and healthy donors and in AML cell lines were quantified by quantitative real-time PCR and Western blotting. The correlation between GADD45g expression and overall survival (OS), coupled with event-free survival (EFS) in patients with AML was analyzed in two gene expression datasets (GSE10358, GSE425-GPL317). Lentiviral vectors over-expressing GADD45g were constructed and transfected into AML cell lines (U937, THP-1 and Molm-13 cell lines). The role of GADD45g over-expression in cell proliferation, colony formation, senescence, apoptosis, cell cycle arrest, differentiation and drug sensitivity of U937, THP-1 and Molm-13 cells were detected by cell counting, colony-forming assay, β-galactosidase staining and flow cytometric analysis of Annexin V/7AAD staining, PI staining and so on, respectively. Results: Expression of GADD45g was dramatically down-regulated in BMMNCs in AML patients and AML cell lines compared to that from healthy donors (all P<0.01). The OS (P<0.05) and EFS (P<0.05) of AML patients with low GADD45g expression were significantly shorter that those of AML patients with higher GADD45g level. Enforced expression of GADD45g could inhibit cell growth and colony formation, promote senescence and apoptosis, induce cell cycle arrest and differentiation and enhance drug sensitivity in AML cell lines (P<0.05 or P<0.01). Conclusion: GADD45g expression was remarkably silenced in marrow tissues of patients withAML andAML cell lines; it showed remarkable and all-around inhibiting effect onAMLcell lines, indicating that GADD45g expression has prognostic value inAML.

Research Progress of Gas6 with Type 2Diabetes Mellitus and Diabetic Complications / 昆明医科大学学报

Growth arrest-specific 6, which is a member of the vitamin K-dependent protein family, is a ligand of TAM (Tyro3, Axl, Mer) belonging to the receptor tyrosine kinase family.Gas6/Axl signaling pathways are involved in pathophysiological processes of cell adhesion, migration, proliferation, apoptosis, platelet aggregation, vascular remodeling, inflammatory and immune responses. In recent years, studies found that Gas6/Axl signaling pathway, Gas6 level and the gene polymorphism of Gas6 were significantly associated with type 2 diabetes, obesity, insulin resistance and diabetic complications.This article reviews the correlation between Gas6 and type 2 diabetes and its complications

Low Nanomolar Cucurbitacin-I Induces G2 / M Phase Arrest and Apoptosis in Gastric Cancer Cells and the Underlying Mechanism / 胃肠病学

Background:Upon inhibition of STAT3 signaling pathway,cucurbitacin-I elicits anticancer effect in various malignancies. However,the anticancer effect and underlying mechanism of cucurbitacin-I in gastric cancer is still elusive. Aims:To explore the effect of low nanomolar cucurbitacin-I on cell proliferation,cell cycle and apoptosis in human gastric cancer cells and the underlying mechanism in vitro. Methods:Human gastric adenocarcinoma cell lines AGS and HGC-27 were treated with cucurbitacin-I at low nanomolar concentration. The anti-proliferative effect of cucurbitacin-I was detected by CCK-8 assay and colony formation assay. Flow cytometry was used to assess the cell cycle and apoptosis. Expressions of cell cycle-related proteins,as well as activation of related pathways such as caspase-3 / PARP apoptotic pathway,STAT3, GADD45α and JNK/ p38 MAPK signaling pathways were determined by Western blotting. Results:Cucurbitacin-I markedly inhibited the growth of gastric cancer cells at low nanomolar concentration by inducing G2 / M phase arrest and apoptosis via a STAT3-independent manner. Furthermore,it was revealed that the anticancer effect of cucurbitacin-I was associated with up-regulation of GADD45α,activation of JNK/ p38 MAPK signaling pathway and the subsequent apoptotic events. Conclusions:The present study provides new insights into the mechanism of anticancer effect of cucurbitacin-I, supporting cucurbitacin-I as an attractive therapeutic drug in gastric cancer.

The efficiency of serum lncRNA GAS5/miR-21 as biomarkers in patients with diabetes and diabetic nephropathy / 中华肾脏病杂志

Objective To elucidate the efficiency lncRNA GAS5 and miR-21 as biomarkers in diabetes mellitus and diabetic nephropathy.Methods The patients were divided into three groups,diabetic nephropathy group (DN group proven by renal biopsy,n=25,14 males and 11 females),diabetes group (DM group,with normal urine albumin creatinine ratio,n=10,4 males and 6 females),and normal control group (NC group,n=9,4 males and 5 females).The expressions of lncRNA GAS5 and miR-21 in serum samples were detected by real-time quantitative PCR.The correlation between serum lncRNA GAS5 and miR-21 expressions and the clinical parameters was analyzed by T-test,Pearson,Spearman test and multivariate linear regression analysis.Differences of lncRNA GAS5 and miR-21 in different groups were analyzed by one-way analysis of variance.The ROC curve was used to analyze the diagnostic efficacy of lncRNA GAS5 and miR-21 in diabetes and diabetic nephropathy.All data were analyzed by SPSS 20.0 and GraphPad software,with P ＜ 0.05 as considered statistically significant.Results (1) The expression of serum lncRNA GAS5 was significantly down-regulated and serum miR-21 was significantly up-regulated in both diabetes mellitus and diabetic nephropathy patients compared to the NC group all (P ＜ 0.05).(2) In DN patients,the expression of serum lncRNA GAS5 was gradually up-regulated along with the increment of 24 h urinary protein.The expression of serum miR-21 was gradually up-regulated along with renal biopsy stage Ⅱb-Ⅲ of DN (P ＜ 0.05).(3)FBG and HbA1c were all negatively correlated with serum lncRNA GAS5 (P ＜ 0.05),and FBG was independently correlated with serum lncRNA GAS5 (P ＜ 0.05).Urine microalbumin,Total cholesterol (TC),Scr,Urea and SBP were all positively correlated with serum miR-21(P ＜ 0.05).Albumin (ALB)and estimated GFR (eGFR) were negatively correlated with serum miR-21(P ＜ 0.05),and ALB was independently correlated with serum miR-21 (P ＜ 0.05).(4) The diagnostic efficiency of serum lncRNA GAS5,miR-21 and lncRNA GAS5/miR-21 as "diagnostic signature" for DM were was good (P ＜ 0.05).(5) The diagnostic efficiency of serum miR-21 and lncRNA GAS5/miR-21 as "diagnostic signature" for DN were was good (P ＜ 0.05).Conclusions (1) Serum lncRNA GAS5 had good diagnostic efficiency in diabetes mellitus.The sensitivity of lncRNA GAS5/miR-21 for diagnosis of diabetes was 85.71％,and specificity was 88.89％.(2) The level of serum miR-21 can be used as a noninvasive diagnostic marker for diabetic nephropathy.

Effects of Gadd45 α-knockdown on human extravillous trophoblast cell migration and invasion functions under hypoxia/re-oxygenation / 上海交通大学学报（医学版）

Objective · To investigate the effects of growth arrest and DNA damage 45 alpha (Gadd45α) on the migration and invasion function of human extravillous trophoblast cells under hypoxia/re-oxygenation (H/R). Methods · Human extravillous trophoblast cells were infected by shRNA lentivirus targeting Gadd45α gene, to knock down Gadd45α gene expression. Then the oxidative stress model of preeclampsia was used in vitro to observe the changes of cell biological functions. The experiments were divided into 4 groups, nontreated group, hypoxia/re-oxygenation group, shRNA Gadd45α+H/R group and shRNA negative control+H/R group. Human villous explant experiments were used to determine the effects of silencing Gadd45α on human extravillous trophoblast cell under oxidative stress. Protein expression of Gadd45α was identified by Western blotting. Changes of cell migration and invasion were detected by transwell migration and Matrigel invasion assay. Gelatin zymography was used to detect the expression of matrix metalloproteinase (MMP) -2/9 in culture medium. Results · Hypoxia/re-oxygenation can increase the expression of Gadd45α in HTR8/SVneo cells and damage the trophoblast cell migration and invasion. Knocking down Gadd45α can increase the activities of MMP2/9, which can increase the cell migration and invasion. Conclusion · Knockdown of Gadd45α gene has promoted cell migration and invasion function of human extravillous trophobalst cells under oxidative stress.

Serum sex hormone and growth arrest-specific protein 6 levels in male patients with coronary heart disease / 亚洲男科学杂志(英文版)

Epidemiological studies have shown a high prevalence of low serum testosterone levels in men with cardiovascular disease. Moreover, the tyrosine kinase receptor Axl, the ligand of which is growth arrest-specific protein 6 (GAS6), is expressed in the vasculature, and serum GAS6 levels are associated with endothelial dysfunction and cardiovascular events. Testosterone regulates GAS6 gene transcription directly, which inhibits calcification of vascular smooth muscle cells and provides a mechanistic insight into the cardioprotective action of androgens. This study was designed to determine the correlation between serum GAS6 and testosterone levels in male patients with coronary heart disease (CHD). We recruited 225 patients with CHD and 102 apparently healthy controls. Serum concentrations of GAS6 and soluble Axl were quantified by an enzyme-linked immunosorbent assay. Levels of high-sensitivity C-reactive protein, testosterone, estradiol, and other routine biochemical markers were also measured. Testosterone decreased from 432.69 ± 14.40 to 300.76 ± 6.23 ng dl-1 (P < 0.001) and GAS6 decreased from 16.20 ± 0.31 to 12.51 ± 0.19 ng ml-1 (P < 0.001) in patients with CHD, compared with control subjects. Multiple linear regression analysis showed that serum testosterone and GAS6 levels were positively associated in male patients with CHD. Alterations in GAS6 levels may influence the development of CHD. Downregulation of GAS6/Axl signaling in the presence of low sex hormone levels during disease progression is a potential mechanism by which GAS6 affects CHD. This study provides novel results regarding the influence of sex hormones on serum GAS6 levels in patients with CHD.

Experimental study onα-Asarone-induced apoptosis in human esophageal carcinoma Eca-109 cell line I by regulating GADD153 and Smac mRNA expression levels / 医学研究生学报

Objective α?Asarone has the effect of relieving cough and asthma as well as sedative, hypnotic and anticonvul?sive function. Our study was designed to explore the effects of apoptosis induced by α? Asarone on human esophageal carcinoma Eca?109 cell line as well as the expression levels of GADD153 and Smac mRNA. Methods Human esophageal carcinoma Eca?109 cells were cultured in vitro, which were divided into blank control group, 5?FU group( 500μg/mL) andα?Asarone group of different dosages ( 25,50,100μg/mL) . After cultivation, MTT method and Annexin V?PI were used to measure cell proliferation rate and apoptosis rate. Expression levels of GADD153 and Smac protein and mRNA were detected by western blotting and real? time quantitative PCR. Results Compared with blank control group, the cell proliferation rates in other groups decreased significantly (P<0.01), and cell apoptosis rate increased significantly ( P<0.01) . Compared with blank control group, the expression levels of GADD153 and Smac pro?tein and mRNA in other groups increased significantly( P<0.05) . Conclusion α? Asarone can inhibit cell proliferation and induce the apoptosis of human esophageal carcinoma Eca?109 by regulating the expression levels of GADD153 and Smac gene.

Expression and role of long non-coding RNA GAS5 in human carcinomas / 国际肿瘤学杂志

Growth arrest special 5 (GAS5) is a common long non-coding RNA (lncRNA).Recent studies find that lncRNA GAS5 is down-regulated in various tumors,such as breast cancer,cervical cancer,lung cancer,gastrointestinal cancers and urologic cancers.GAS5 is closely related to the development,metastasis and prognosis of cancers,which becomes a new hotspot in cancer research.

Effect of Taurine on the expression of glucose regulated protein 78 and growth arrest and DNA damage - in-ducible protein 34 in zebrafish larvae brain after hypoxia/reoxygenation brain injury / 中华实用儿科临床杂志

Objective To investigate the expression of endoplasmic reticulum stress - related factors, glucose - regulated protein 78(GRP78)and growth arrest and DNA damage - inducible protein 34( GADD34)and neuronal apoptosis in the brain of zebrafish larvae after hypoxia/ reoxygenation brain injury,and the neuroprotective role of Taurine. Methods The 5 day post - fertilization zebrafish larvae were randomly assigned to 3 groups:control group, hypoxia/ reoxygenation model group(model group)and Taurine treatment group(Taurine group). According to the dif-ferent time points for observation,each group was subdivided into 5 subgroups(1 h,3 h,6 h,24 h,48 h)with 100 ze-brafish larvae each. The pathological changes in the brain tissues and cell apoptosis were detected by fluorescence ter-minal deoxynucleotidyltransferase - mediated dUTP nick end - labeling( TUNEL). The expression of GRP78 and GADD34 mRNA in the brain of zebrafish larvae were detected by real - time quantitative reverse transcription PCR (qRT - PCR). The changes in GRP78 and GADD34 protein were detected by Western blot. Results (1)TUNEL:apoptosis index(AI)was increased after hypoxia/ reoxygenation,and reached the peak at 3 h in model group,the AI in Taurine group was decreased compared with that in the model group at the same time point(1 h:22. 83 ± 1. 80 vs 30. 18 ± 1. 81,3 h:23. 22 ± 2. 46 vs 42. 97 ± 4. 01,6 h:16. 80 ± 1. 69 vs 22. 97 ± 1. 91,all P ﹤ 0. 05).(2)qRT -PCR:the expression of GRP78 and GADD34 mRNA was increased at 1 h after hypoxia/ reoxygenation,and reached the peak at 3 h in the model group,the expression in Taurine group was decreased compared with that in the model group at the same time point(GRP78 mRNA:1 h:2. 35 ± 0. 13 vs 5. 36 ± 0. 35,3 h:3. 08 ± 0. 33 vs 4. 27 ± 0. 52,6 h:1. 57 ± 0. 12 vs 3. 00 ± 0. 13,all P ﹤ 0. 05;GADD34 mRNA:1 h:5. 14 ± 0. 55 vs 7. 45 ± 0. 67,3 h:2. 79 ± 0. 58 vs 5. 83 ± 0. 51,6 h:1. 79 ± 0. 22 vs 3. 67 ± 0. 30,all P ﹤ 0. 05).(3)Western blot:the expression of GRP78 and GADD34 pro-tein was increased at 1 h,reached the peak at 6 h,but it was decreased in Taurine group(GRP78 protein:1 h:1. 12 ± 0. 11 vs 1. 37 ± 0. 13,3 h:0. 79 ± 0. 11 vs 1. 25 ± 0. 10,6 h:0. 55 ± 0. 10 vs 1. 52 ± 0. 14,all P ﹤ 0. 05;GADD34 pro-tein:1 h:0. 92 ± 0. 11 vs 1. 11 ± 0. 13,3 h:0. 96 ± 0. 11 vs 1. 52 ± 0. 09,6 h:0. 76 ± 0. 05 vs 1. 89 ± 0. 06,all P ﹤0. 05).(4)The expression of GRP78 and GADD34 protein was positively correlated with AI in the model group(r =0. 53,0. 56 respectively,all P ﹤ 0. 05). Conclusion One of neuroprotective mechanisms of Taurine against hypoxia/reoxygenation brain injury may down - regulate GRP78 and GADD34 expression.

Influence of edaravone on growth arrest and DNA damage-inducible protein 34 expression following focal cerebral ischemia-reperfusion in rats

Objective: To investigate the influence of edaravone on the expression of growth arrest and DNA damage-inducible protein 34 (GADD34). Methods: A total of 108 healthy male Sprague-Dawley rats were randomly divided into sham operation group, model group and edaravone group (36 cases for each group). Transient focal cerebral ischemia was induced by middle cerebral artery occlusion for 2 h followed by reperfusion in Sprague-Dawley rats. Then, GADD34 expression was measured with immunohistochemistry at different time-points after reperfusion in the peri-infarct regions of all rats. Results: The GADD34 expression was detected in the peri-infarct regions of rats 1 h after reperfusion, which reached its peak 24 h after reperfusion. And edaravone could significantly down-regulate the GADD34 expression. Conclusions: Edaravon could down-regulate GADD34 expression, which suggests that edaravone may exert an important function in inhibiting endoplasmic reticulum stress reaction by scavenging free radicals in the upper stream.

Assay of serum growth arrest-specific gene product 6 and stromal cell-derived factor isoform-1 in patients with diabetic retinopathy / 中华实验眼科杂志

Background Diabetic retinopathy (DR) is a progressive vision-threatening complication of diabetes mellitus (DM),but its pathogenic mechanism is still unclear.Researches showed that growth arrest-specific gene product 6 (Gas6) /TAM system participates in pathogenesis and development of DR,and stromal-derived factors (SDF) vary in 2-type DM patients.However,whether Gas6/TAM and SDF-1 are associated with DR is below understood.Objective This study was to determine the relationship between the staging of DR and the levels of serum Gas6,SDF-1α and SDF-1β in DM patients.Methods A prospective cohord study was designed in this study.Ninety 2-type DM patients were included in the 3rd Affiliated Hospital of Southern Medical University Hospital from January to August in 2013.The patients were grouped into the non-diabetic retinopathy (NDR) group,background DR group (BDR) and proliferative DR (PDR) group,with 30 for each group.Thirty normal volunteers were enrolled in the same hospital and same period.The periphery blood 2 ml was collected from all the subjects under the consent inform.The levels of serum Gas6,SDF-1α and SDF-1β were assayed by ELISA,and leukocytes,neutrophils,plasma triglycerides (TG),total cholesterol (CHOL),high density lipoprotein cholesterol (H DL-C) and low density lipoprotein cholesterol (LDL-C) levels were detected and compared among the 4 groups.The correlations of serum Gas6,SDF-1 α and SDF-1 β changes with blood inflammatory cells and blood lipid were analyzed.Results The plasma CHOL concentrations were 4.93(4.14,5.44),5.02(4.35,5.69),4.54(3.85,5.93) and 5.99(5.11,6.89)mmol/L in the normal control group,NDR group,BDR group and PDR group,respectively,and the blood CHOL concentrations were significantly higher in the PDR group than those of the normal control group,NDR group,BDR group (P =0.002,P =0.007,P =0.006).White blood cell counts in the normal control group,BDR group were higher than those of the PDR group (P =0.034,P =0.015),neutrophil counts in the BDR group were higher than those of the PDR group (P =0.024),HDL-C in the NDR group was higher than that in the PDR group (P =0.032).LDL-C in the PDR group was higher than that in the normal control group.Compared with the normal control group,serum Gas6 levels were significantly lower in the NDR group and BDR group (P =0.048,P =0.006),and the serum Gas6 level showed an insignificant increase in the PDR group in comparison with BDR group (P =0.297).Serum SDF-1α levels in the PDR group was significantly higher than that in the BDR group (P =0.033) ;serum SDF-1β levels in the PDR group,BDR group were significantly higher than that in the NDR group (P =0.011,P =0.008) and normal control group (P =0.030,P =0.002).Weaker positive correlation was observed between the serum Gas6 and CHOL,TG,LDL-C levels (r=0.285,r=0.200,r=0.241,all at P＜0.05),between SDF-1α and SDF-1β (r=0.190,P＜0.05) as well as between SDF-1β and white blood cell (r=0.183,P＜0.05).Serum Gas6 served as dependent variable,while white blood cell,neutrophil,CHOL,TG,HDL-C,LDL-C,SDF-1α,SDF-1β served as independent variables,multiple stepwise regression analysis showed Gas6 =170.791 + 5.283CHOL (F =5.021,P =0.027).Conclusions Serum Gas6,SDF-1α and SDF-1β probably participate in the development of DR in 2-type diabetic patients.Gas6,SDF-1 α,SDF-1 β may play roles by affecting blood glucose level,angiogenesis,inflammatory cells and blood lipid metabolism.