Gene Expression of GSK3, TGF ?-1 and PI3 Kinase in Oral Squamous Cell Carcinoma – A Real Time PCR Based Approach

Background: Oral squamous cell carcinoma (OSCC) is the most predominant type of oral cancer which has a poor prognosis, with 5-year survival rates less than 50%. Clinical characteristics such as tumor position, TNM classification and method of treat- ment, as well as histological grades have all been studied as OSCC prognostic factors but evaluating the genetic expression is the evolving trend in early diagnosis. Aim: To compare the gene expression of TGF-?-1, GSK3, Pi3 kinase in OSCC and normal tissue samples and to correlate the expression levels of these molecules with the pathological grading and survival in OSCC patients. Also to understand the role of GSK3 in Pi3 kinase pathway and TGF-? signaling pathway in OSCC progression thereby attempting targeted therapy in OSCC patients. Materials and Methods: 10 OSCC samples as well as normal healthy samples were collected and RNA isolation was done us- ing RNA easy kit from Qiagen (Valencia, CA), and thensubjected to cDNA synthesis using Human TGF-?1, Human GSK3? and Human Pi3 kinase primers. Real time PCR was performed using gene specific primers at 40 cycles. The results were retrieved, tabulated and analyzed. Results: The current research results revealed that there were up regulation of mRNA expression in GSK3, TGF ?-1 and Pi3 kinase in OSCC patients than in healthy individuals. On comparison, Pi3 kinase showed highest mRNA expression levels than GSK3 and TGF ?-1. Conclusion: The expression of GSK3 and its role in activation of Pi3 kinase pathway plays a crucial role in progression of oral cancer and targeting GSK3?could be a novel and targeted approach for treating OSCC.

Transforming growth factor-1 induces angiogenesis in vitro via VEGF production in human airway smooth muscle cells.

Increase in size and number of bronchial blood vessels as well as hyperaemia are factors that contribute to airway wall remodelling in patients with chronic airway diseases, such as asthma and chronic obstructive pulmonary diseases (COPD). Expression of transforming growth factor 1 (TGF-1), a multifunctional cytokine as well as vascular endothelial growth factor (VEGF), a key angiogenic molecule, has been shown in the inflammed airways in patients with chronic airway diseases. TGF-1 has been implicated in the regulation of extracellular matrix, leading to airway remodelling in patients with chronic airway diseases. However, the role of TGF-1 in regulating VEGF expression in patients with chronic airway diseases, as well as the underlying mechanisms are not yet well established. We investigated whether TGF-1 stimulates VEGF expression in vitro and hence could influence vascular remodelling. Cultured human airway smooth muscle cells (HASMC) were serum deprived for 60 h before incubation with 5ng/ml of TGF-1 for different time points. Control cells received serum-free culture medium. TGF‑1, treatment resulted in time dependent HASMC cell proliferation with maximal values for DNA biosynthesis at 24 h and cell number at 48 h. Northern blot analysis of VEGF mRNA expression showed increased levels in cells treated with TGF-1 for 4 to 8 h. TGF-1 also induced a time-dependent release of VEGF proteins in the conditioned medium after 48 h of treatment. Furthermore, the ability of HASMC-released VEGF proteins to induce human umbilical vein endothelial cells proliferation was inhibited by VEGF receptor antagonist, confirming that TGF-1 induced VEGF was biologically active. We conclude that TGF-1 in addition to an extracellular matrix regulator also could play a key role in bronchial angiogenesis and vascular remodelling via VEGF pathway in asthma.

Effect of 1,25-dihydroxyvitamin D_3 on renal expression of TGF-1 and HGF in type 1 diabetic rats / 重庆医科大学学报

Objective:To explore the effects of 1,25-dihydroxyvitamin D3 on renal expression of TGF-?1 and HGF in type 1 diabetic rats.Methods:30 Sprague-Dawley rats were randomized into 3 groups:normal control rats(group NC),diabetic rats(group DM ) and diabetic rats treated with 1,25-dihydroxyvitamin D3(group DC).Kidney-to-body weight ratio(KW/BW),mean glomerular volume(MGV),fractional mesangial area(FMA),urinary albumin excretion rate(UAER)and serum creatinine(Scr)were measured after 12 weeks of treatment,while transforming growth factor-betal(TGF-?1) expression in kidney cortex were observed by immunohistochemistry.The expression levels of TGF-?1 mRNA and HGF mRNA were detected by reverse transcription and polymerase chain reaction(RT-PCR) at the end of the experiment.Results:UAER,Scr,MGV,FMA and KW/BW ratio were significantly higher in the diabetic rats than those in the normal control rats(P

Effects and Mechanisms of PNS on Myocardial Fibrosis in Chronic Viral Myocarditic Mice / 浙江中医药大学学报

[Objective] To study the mechanisms and effects of panax notoginsenosides(PNS)on myocardial fibrosis in chronic viral myocarditic(VMC)mice.[Methods]The model mice with VMC at chronic status were established by repetitive infection of Coxsckievirus B3m(CVB3m).Heart changes of myocardium and collagen hyperplasia were observed by HE and VG stain.Collagen volume fraction(CVF)and perivascular circuferential area(PVCA)were examined by pathological examination with computed processing.Myocardium TGF-?1 was detected by immunohistochemical method and mRNA expression of TGF-?1 was analyzed by Reverse transcription polymerase chain reaction(RT-PCR)method.[Results]Marked myocardial fibrosis was observed in chronic VMC model group.Compared with blank group,the index of CVF and PVCA was increased significantly(P

Expression of protein vEGF and TGF-?1 in the process of cicatrization of incised wounds on rat skin / 重庆医科大学学报

Objective:To investigate whether the expression of protein VEGF and TGF-?1 can be used as an indextoestimate injuryage in the field of forensic medicine by approaching the temporal relation of VEGF and TGF-?1 expression with injury in the healing process of the incised wounds on rat skin.Methods:Experimental rat models were established by incisions made on the rat skin.the rat models were divided randomly into three groups:the experimental group,the normal group,and the postmortem injured group.The expression of VEGF and TGF-?1 were observed with Immunohistochemical techniques in their process of cicatrization.Results:In the experimental group,the expression of VEGF and TGF-?1 were higher than those in the normal control and postmortem injury group(P

Comparison of transforming growth factor-?_1 expression and therapeutic effects on tibia and fibular fracture with two different fixation methods / 重庆医科大学学报

Objective:To detect the expression of TGF-?1 in tibia and fibula fracture with two methods of compression plate fixation and intramedullary nail fixation,and to evaluate their therapeutic effects.Methods:117 patients with non severe tibia and fibula fractures were separated into two groups.One group received pressurized plates fixation for treatment,the other received intramedullary nail fixation.On 1,7,30 and 90 days after surgery,TGF-?1 was detected by ELISA,and on 30 days after surgery the clinical efficacy were also assessed. Results:The TGF-?1 content increased on 7 days as compared with that on 1 days after surgery,and the TGF-?1 content gradually decreased at on 30 to 90 days after surgery.The TGF-?1 content in compression plate fixation group were lower than that in intramedullary nailing group every points of four days(P

Changes of PI3K/AKT expression in renal tubulointerstitial fibrasis of unilateral ostruction rats / 基础医学与临床

Objective To observe the effect of PI3K/AKT signal passageway on renal tubulointerstitial fibrogenesis of UUO rats.Methods Thirty Sprague Dawley rats were divided into two groups,the sham-operated group and the uuo model group.The model group's left ureter was dissociated to be obstructed.The sham-operated group was dissociated left ureter only without obstruction.Five rats of each group were killed respectively at 5,10,15(days after) surgery.Histological changes of renal tubular interstitium was observed by HE and Masson staining.Immunohistochemistry was performed on left renal tubular interstitium for AKT1,phospho-AKT1,TGF-?1 in renal(tubulointerstitum) at each time point.Results AKT1,phospho-AKT1and TGF-?1 were faintly stained in the sham-operated kidneys.Immunostaining of AKT1,phospho-AKT1 and TGF-?1 in renal tubulointerstitium increased(progressively) starting from 5 to 15 days after UUO(vs sham-operated group,P

Relationship of angiogenesis and clinical characteristics of GA-secreting pituitary adenomas / 基础医学与临床

Objective To investigate MVD, TSP-1, TGF-?1 expression in GH-Secreting Pituitary Adenomas by immunohistochemistry, and to correlate data with clinical characteristics.Methods The protein expression of TSP-1, TGF-?1 in 48 surgical specimens (21 invasive cases; 27 non-invasive cases) of pituitary adenomas was measured using immunohistochemical method. The relationship between the expression and clinical properties was examined. MVD was measured by detecting CD34.Results Compared with the noninvasive group, no difference of expression of CD34(t=2.257; P=0.083) was observed. The expression of TSP-1 in invasive group was low. The expression of TGF-?1 was higher in invasive cases than that in noninvasive ones. The expression of TGF-?1 had positive correlations with MVD; but there was no correlation between the expression of CD34 and the invasion of pituitary adenomas. In addition, MVD count was not associated with the expression of TSP-1. Size, sex or rate of recurrence did not influence MVD and TSP-1 expression. Conclusion MVD values do not necessarily represent angiogenesis in pituitary adenomas. TGF-?1 may increase MVD, and TSP-1 does not affect MVD in pituitary adenomas and angiogenesis may be regulated by other pathway.

Adipose tissue derived stem cells transferred with TGF-?_1 gene and differentiated into chondrocytes / 中国矫形外科杂志

[Objective]To induce TGF-?1 gene which can increase ECM synthesis into adipose tissue derived stem cells(ADSCs) by employing gene transfer techniques and observe whether TGF-?1 gene could expresse continuously and whether type II collagen and aggrecan are synthesized in order to provide experimental data for constructing tissue-engineering cartilage. [Method]ADSCs were transferred with TGF-?1 gene ,TGF-?1,FN,ColⅡ and aggrecan were detected with RT-PCR and TGF-?1 protein was detected with Western-blot.[Result]RT-PCR demonstrated that the expression of TGF-?1,FN,ColⅡ and aggrecan in the TGF-?1 gene transferred groups increased more evidently than non-gene groups and control groups (P

Expression and signififcance of IGF-Ⅰ TGF-?1 in human mesenchymal stem cells after exposured to extracoporeal shock wave / 中国矫形外科杂志

[Objective]To demonstrate mechanism of ESW in curing osteogetic disorders,we studied expression of some osteogenetic factors in human mesenchymal stem cells(hMSCs)when exposed to ESW.[Method]After success in marrow aspiration,isolation and obtainment optimal experimental energy,a dose of 5kV and 100 times of ESW was applied to hMSCs of passage 1.The expression of IGF-Ⅰ and TGF-?1 were examined by immunocytochemical staining.[Result]The cytochemical staining results showed that expression of IGF-Ⅰ and TGF-?1 appeared at different passage of hMSCs after ESW intervention.Appearance of IGF-Ⅰ was earlier than TGF-?1 which didnt express until passage 7.At the same interval,the expression of IGF-Ⅰ and TGF-?1 in control group difference is lower than ESW group,respectively(P

Intervertebral disc cell therapy for regeneration:mesenchymal stem cell cultivation in vitro and intervention in TGF-?1 implantation in rabbit intervertebral discs / 中国矫形外科杂志

[Objective]To evaluate whether transplanted marrow mesenchymal stem cells interfered in TGF-?1 can differentiate to nucleus pulposus cells and increase the amount of proteoglycan and collagenase Ⅱ content in intervertebral discs.[Method]We used an in vivo model to investigate the feasibility of marrow mesenchymal stem cells that cultured in vitro and interfered in TGF-?1 delivery,retention,and survival in the degeneratived disc space.In 2,4,6,8 weeks we used immunohistochemical staining to determine the change of collagenase Ⅱ content;spectrophotometry to determine the change of amount of proteoglycan with Phlorglucinol;the experiment date were analyzed by SPSS 11.5 soft ware.[Result]We found MSCs could maintain viability and proliferate within the rabbit inter vertebral disc.The amount of proteoglycan and collagen Type Ⅱ content of the intervertebral in matrix synthesis in the experiment group was increased in 8 weeks.We found no changes in the modle group.[Conclusion]Our data suggest that transplanted marrow mesenchymal stem cells in vivo can survive and increase proteoglycan and collagen Type Ⅱ amount interfered in TGF-?1 in some periods,which support its potential use as a treatment of intervertebral disc degeneration.

Association of the promoter~(-509)C>T polymorphism of transforming growth factor-?1 with lumbar disc disease / 中国矫形外科杂志

T polymorphism of TGF-?1 gene related with the degree of L_(4?5) and L_5S_1 disc degeneration.

The Effect of Cytokine on Myocardial Fibrosis of Chronic Viral Myocarditis and the Interventional Treatment of QingXinⅡ / 浙江中医药大学学报

[Objective]To study the effect of TGF?1 on the expression of fibronectin and ?1 integrin receptor in chronic viral myocarditis of myocardial fibrosis induced by repeated virus infection and the interventional effect of QingXin.[Methods] Establish mice model of myocardial fibrosis in chronic stage of viralmyocarditis,then divide the mice model into model group,captopril group,QingXinⅡ high dose,medium dose and low dose groups.The normal group and model group are given saline through stomach perfusing,as well as the treated group are given respectively captopril and QingXinⅡ high,medium and low dose in the same way.After the treatment,hearts are collected,CVB3-RNA are detected by RT-PCR,TGF?1 and FN are detected by immunohistochemical technique,integrin?1 are analyzed by immunofluorescence staining-confocal laser scanning.[Results] After having done the model,TGF?1,integrin?1,FN has increased obviously;virus can be detected in themyocardium by RT-PCR;after the intervention of QingXinⅡ,TGF?1,integrin?1,FN has decreased,virus has reduced in treated group.[Conclusion] Virus,TGF?1,integrin?1 and FN have important effects on the formation ofmyocardial fibrosis;QingXinⅡ has the function of antivirus and anti-myocardial fibrosis.

Comparison Study between Curcumin and Enalapril Treatment in Unilateral Ureteral Obstruction Rats / 中国中医药信息杂志

Objective To investigate the effect of curcumin on renal interstitial fibrosis following unilateral ureteral obstruction (UUO), and compare the effects of Curcumin with that of Enalapril. Method UUO rat model was used. Male Wistar rats were randomly divided into four groups with 8 rats each:shame-operated group (0.9% saline solution), model group (dimethyl sulphoxide, DMSO), Curcumin treatment group [curcumin 100 mg/(kg?d)], and Enalapril treatment group [enalapril 10 mg/(kg?d)]. All rats were killed 4 w after surgery. The degree of tubulointerstitial fibrosis was scored by HE and Masson staining. The sites and levels of protein expression of TGF-?1, CTGF and TIMP-1 were detected by immunohistochemistry staining. Result Compared with those of shame-operated group, the blood urea nitrogen level was significantly increased (P 0.05). Conclusion Pharmacological effects of Curcumin or Enalapril on unilateral ureteral obstruction rats are quite similar. These effects may result from the down-regulation of TGF-?1, CTGF and TIMP-1 expression in kidney tissue.

Effects of Puerarin on Renal Advanced Glycosylation End Products and Expression of TGF-?_1 in Diabetic Rats / 中国中医药信息杂志

Objective To investigate the effects of puerarin on renal advanced glycosylation end products (AGEs), expression of transforming growth factor-?1 (TGF-?1) and pathomorphism in diabetic rats. Methods Rats were randomly divided into following groups:normal control rats (Group CN), diabetic rats (Group DM) and diabetic rats treated with puerarin (Group DP). After Puerarin was given to STZ-induced diabetic rats for 10 weeks, blood glucose, urine protein, glomerular basement membrance thickness (GBMT) and profile of kidney hypertrophy in diabetic rats were measured and analysed. The renal pathologic changes were observed by light microscopy and electron microscopy. Renal AGEs content was determined by expose time of fluorescence microscopy. The expression of TGF-?1 was examined by immunohistochemical methods. Results Urine protein, GBMT and profile of kidney hypertrophy were significantly increased in diabetic rats as compared with control animals (P

Matrine Compoand Injection Interrening the Liver Fibrosis Rat Induced by Tetrachloride / 医学研究杂志

Objective To examine the change of TGF-?1 in the bloods of rats which were induced by tetrachoride during the different times and to explain the mechanism of the change.MethodsThe experimental liver fibrosis induced by tetrachloride in rats was treated by matrine compound injection.Serum TGF-?1 and pathological changes in liver tissue were observed in control group,model group and therapy group during the time of the 4th,6th and 9th week.ResultsThere was an obvious change of TGF-?1 in the blood in different time and different groups.ConclusionMatrine compound injection has an antifibrogenetic action and TGF-?1 can reflect the change of liver fibrogenesis of different time as an index.

Effects of Traditional Chinese Medicine Ruanganyin on TGF-1/Smad Signaling Pathway in Carbon Tetrachloride-induced Hepatic Fibrosis of Rats / 浙江中医药大学学报

[Aim]To study the effects of Ruanganyin on TGF-1,Smad3,Smad7 expressions of the experimental hepatic fibrosis induced by carbon tetrachloride (CCl4) in rats,and to explore its anti-fibrotic molecular mechanism.[Method] Forty healthy Wistar rats were randomly divided into normal control group,vacant control group,CCl4-induced hepatic fibrosis group,and Ruanganyin treated group.The rats in normal group were treated with normal saline by intragastrical irrigation,and the other groups were subcutaneous injected with CCl4 to establish hepatic fibrosis model.The histological changes of the liver tissues were examined by pathological examination.The protein expression of transforming growth factor beta 1 (TGF-1) and Smad3,Smad7 were measured by immunohistochemical technique.[Result] In comparison with those inmodel group,the levels of TGF-1 were significantly decreased after treatment with Ruanganyin.Meanwhile,the levels of Smad3 was also down-regulated.However,the level of Smad7 expression was significantly up-regulated.The pathological changes of liver tissues in Ruanganyin groups were also markedly alleviated.[Conclusion] Ruanganyin has significant inhibitory effects on CCl4-induced hepatic fibrosis in rats,and its mechanism may be associated with the inhibition of TGF-1 expression,down-regulation of Smad3 expression and up-regulation of Smad7 expression.

The Effects of Low Molecular Weight Heparin on Serum Level of PCⅢ and Type IV Collage and TGF-?_1 / 医学研究杂志

Objective To study effects of low molecular weight heparin on liver fibrosis and the serum levels of TGF-?_1 in patients with chronic hepatitis B virus.Methods 45 patients with chronic hepatitis B virus were randomized into control group(the routine strategy)and trial group(the routine strategy + low molecular weight heparin).The period of treatment is 3 weeks.Serum hepatic fibrosis indices before and after heparin treatment were examined by RIA,the levels of serum TGF-?_1 before and after heparin treatment were examined by ELISA.Results Hepatic functions became significantly better in trial groups,serum PⅢP and type IV collage levels and the levels of serum TGF-?_1 in trial group decreased significantly after treatment.Conclusions The mechanism of anti-fibrosis action of LMWH may inhibited production of TGF-?_1 in patients with HBV.

Research on the Relationship Between the Changing Law of the Cytokines and Tumor-related Fatigue in Gastric Carcinoma / 医学研究杂志

Objective To study the relationship between the changing law of cytokine gene expression profile and tumor-related fatigue and to verify the preliminary theory conclusion that the pathogenesis of tumor-related fatigue may be due to the up-regulation of the TNF,TGF-?,etc.Methods The changing characteristics of gene expression profile of cytokines were studied in peripheral blood mononuclear cells samples with gene chip.Results The gene expression level of TNF-? and TGF-?1 was up regulated.Conclusions The experimental results conform to the theory research conclusion and show that the pathogenesis of tumor-related fatigue due to the up-regulation of the gene expression level of TNF and TGF-?.

A study on effect of Decorin in experimental pulmonary fibrosis / 中国实用内科杂志

Objective To investigate the protective effect and possible mechanism of Decorin on pulmonary fibrosis.Methods A total of 90 healthy Wistar rats were randomly divided into three groups:group A served as control group,group B was injected BLMA5 through trachea,group C was injected BLMA5 and received treatment of Decorin.Ten rats in each group were killed 7、14、28 days after intratrachael instillation.TGF-?_1 expression was measured by immunohistochemical technique.The degree of alveolitis and fibrosis were measured by HE staining.The contents of hydroxideproline were measured in the homodenate of the rat lung.Results The expressions of TGF-?_1 were increased gradually in group B;in group C the expressions were decreased significantly than group B(P