LINC01268 promotes epithelial-mesenchymal transition,invasion and metastasis of gastric cancer via the PI3K/Akt signaling pathway and targeting MARCKS

BACKGROUND Long non-coding RNAs(lncRNAs)with differential expression characteristics have been found to be closely related to the tumorigenesis and development of gastric cancer(GC),but their specific mechanisms and roles still need to be further elucidated.AIM To investigate the expression of LINC01268 in GC and its mechanism of affecting GC progression.METHODS Real-time quantitative polymerase chain reaction was used to detect the expression of LINC01268 in GC tissues,cell lines and plasma.The Kaplan-Meier method was used to evaluate the value of LINC01268 in the prognostication of GC patients.An receiver operating characteristic curve was constructed to evaluate the value of LINC01268 in the diagnosis of GC.Transwell migration and invasion assays and wound healing assays were used to confirm the effect of LINC01268 on the invasion and migration of GC cells.The regulatory relationship between LINC01268 and myristoylated alanine rich protein kinase C substrate(MARCKS),the PI3K/Akt signaling pathway,and the epithelial-mesenchymal transition(EMT)process in GC was demonstrated by western blot analysis.RESULTS The expression of LINC01268 was increased in GC tissues and cell lines.The expression level of LINC01268 was significantly correlated with lymph node metastasis,TNM stage,and tumor differentiation in patients with GC.Over-expression of LINC01268 indicated a poor prognosis for patients with GC,and it had a certain auxiliary diagnostic value for GC.In vitro functional experiments proved that the abnormal expression of LINC01268 further activated the PI3K/Akt signaling pathway and promoted EMT by targeting and regulating MARCKS and ultimately promoted the invasion and metastasis of GC.CONCLUSION This study elucidates that LINC01268 in GC may be an oncogene that further activates the PI3K/Akt signaling pathway and EMT by targeting and regulating MARCKS,and ultimately promotes the invasion and metastasis of GC.LINC01268 may be a potential effective target for the treatment of GC.

Clean-DM1, a Korean Polyherbal Formula, Improves High Fat Diet-Induced Diabetic Symptoms in Mice by Regulating IRS/PI3K/AKT and AMPK Expressionsin Pancreas and Liver Tissues

Objective:To investigate the effects of Clean-DM1(C-DM1),a polyherbal formulation of Radix Scrophulariae,Radix Astragali,Rhizoma Atractylodis,and Radix Salviae Miltiorrhizae,on high-fat diet(HFD)-induced diabetes mice.Methods:The information about active components of C-DM1 extract and molecular mechanism was obtained from network pharmacology analysis.Main compounds of C-DM1 extract by high performance liquid chromatography-mass spectrometry(HPLC-MS)analysis were conducted for quality control.For in vivo study,mice were induced diabetes by HFD for 12 weeks.The mice in the normal group(Nor)were maintained with a regular diet and treated with saline by gavage.The HFD model mice were randomly divided into 3 groups,including a HFD diabetic model group,a C-DM1 extract-administered group(C-DM1,500 mg/kg),and metformin-administered groups(Met,500 mg/kg),8 mice in each group.Food intake,body weight(BW),and fasting blood glucose(FBG)levels were recorded weekly for 4 weeks.After 4 weeks of treatment,alanine aminotransferase(ALT),aspartate aminotransferase(AST),blood glucose,low-density lipoprotein cholesterol(LDL-C)were determined using an automated clinical chemistry analyzer,and homeostatic model for assessing insulin resistance(HOMA-IR)levels and oral glucose tolerance test(OGTT)were detected.The histopathological changes of liver and pancreatic tissues were observed by hematoxylin-eosin staining.Insulin receptor substrate(IRS)/phosphatidylinositol 3 kinase(PI3K)/protein kinase B(AKT)and adenosine 5'-monophosphate-activated protein kinase(AMPK)expressions in liver and pancreas tissues were detected by Western blot analysis.Results:HPLC-MS identified dihydroisotanshinone,dihydroisotanshinone I,cryptotanshinone,harpagoside,and atractyloside A in C-DM1 extract.The administration of C-DM1 extract significantly decreased body weight,calorie intake,and the levels of blood glucose and insulin in the diabetic mice(P<0.05 or P<0.01).The C-DM1 extract administration improved the impaired glucose tolerance and insulin resistance in the diabetic mice and significantly decreased the levels of LDL-C,ALT and AST(P<0.01).The C-DM1 extract inhibited the histopathological changes of fatty liver and hyperplasia of pancreatic islets in the diabetic mice.The C-DM1 extract significantly increased the phosphorylation of IRS,AKT,and AMPK and the expression of PI3K in pancreas and liver tissues(P<0.05 or P<0.01),which was consistent with the analysis results of network pharmacology.Conclusion:C-DM1 extract improved diabetes symptoms in longterm HFD-induced mice by regulation of IRS/PI3K/AKT and AMPK expressions in pancreas and liver tissues,suggesting that C-DM1 formulation may help prevent the progression of T2DM.

Crk-associated substrate,vascular smooth muscle and hypertension

Hypertension is characterized by vascular smooth muscle constriction and vascular remodeling involving cell migration,hypertrophy and growth.Crk-associated substrate(CAS),the first discovered member of the adapter protein CAS family,has been shown to be a critical cellular component that regulates various smooth muscle functions.In this review,the molecular structure and protein interactions of the CAS family members are summarized.Evidence for the role of CAS in the regu-lation of vascular smooth muscle contractility is pre-sented.Contraction stimulation induces CAS phosphor-ylation on Tyr-410 in arterial smooth muscle,creating the binding site for the Src homology(SH)2/SH3 protein CrkII,which activates neuronal Wiskott-Aldrich syn-drome protein(N-WASP)-mediated actin assembly and force development.The functions of CAS in cell migra-tion,hypertrophy and growth are also summarized.Abelson tyrosine kinase(Abl),c-Src,focal adhesion kinase(FAK),proline-rich tyrosine kinase 2(PYK2),pro-tein tyrosine phosphatase-proline,glutamate,serine and threonine sequence protein(PTP-PEST)and SHP-2 have been documented to coordinate the phosphorylation and dephosphorylation of CAS.The downstream signaling partners of CAS in the context of cell motility,hyper-trophy,survival and growth are also discussed.These new findings establish the important role of CAS in the modulation of vascular smooth muscle functions.Furthermore,the upstream regulators of CAS may be new biologic targets for the development of more effective and specific treatment of cardiovascular diseases such as hypertension.

Effect of Shenzhu Tiaopi granule(参术调脾颗粒) on hepatic insulin resistance in diabetic Goto-Kakizakirats via liver kinase B1/adenosine 5’-monophosphate/mammalian target of rapamycin signaling pathway

OBJECTIVE: To observe the therapeutic effect of Shenzhu Tiaopi granule(参术调脾颗粒, STG) on insulin resistance(IR) in the liver of diabetic Goto-Kakizaki(GK) rat and investigate underlying mechanisms.METHODS: Ten 12-week-old male Wistar rats were assigned as normal control(NC) group, while 4012-week-old male specific-pathogen-free GK rats were randomly divided into four experimental groups, 10 diabetic rats each. Animals were fed with a normal diet. Fasting blood glucose(FBG), water intake, and body weight were recorded during6 weeks of daily single-dose treatment: STG low-dose group, 4.5 g/kg(STG-L);STG high-dose group,9 g/kg(STG-H);metformin group, 0.1 g/kg(MET);model control(MC) and NC groups, equal volume of 0.9% Na Cl solution. The serum fasting insulin(FINS), C-Peptide and IR index(HOMA-IR) were detected every 2 weeks during treatment and glucose tolerance was measured in the 3 rd day before the material was taken. After the 6-week STG treatment, Liver tissues were processed for hematoxylin-eosin staining to perform light microscopy analysis and for assessing expression and distribution of insulin receptor substrates(IRS-1) and glucose transporter(GLUT-4) by immunohistochemistry analysis. Expression levels of liver kinase B1(LKB1)/adenosine 5’-monophosphate-activated protein kinase(AMPK)/mammalian target of rapamycin(mTOR) pathway proteins, including LKB1, phospho-AMPK(p-AMPK)/AMPK, phospho-mTOR(p-mTOR)/mTOR, and ribosomal protein S6 kinase polypeptide 1(S6 K1),were detected by Western blotting.RESULTS: STG significantly reduced the FBG level and liver fat deposition in diabetic GK rats. After STG treatment completion, FINS, HOMA-IR, C-Peptide and area under blood glucose curve(AUC)were lower in STG groups than in the MC group, indicating that IR was reduced and liver fat lesions were resolved. In liver tissues, STG groups displayed significantly higher IRS-1 and GLUT-4 expression than the MC group, along with increased LKB1 and p-AMPK/AMPK expression and decreased p-mTOR/mTOR and phospho-S6 K1 expression, suggesting that STG stimulated LKB1 activation of AMPK and suppressed themTOR/S6 K1 downstream pathway.CONCLUSION: Growing GK rats developed hepatic IR, but STG treatment significantly improved hyperglycemia and IR and resolved hepatic fatty lesions.Interestingly, STG treatment stimulated the expression of IRS-1 and GLUT-4 in the liver of diabetic GK rats, indicating a potential involvement in the regulation of the LKB1/AMPK/mTOR signaling pathway.

Regulation of TLR7/9 signaling in plasmacytoid dendritic cells

Plasmacytoid dendritic cells(pDCs),also known as type I interferon(IFN)-producing cells,are specialized immune cells characterized by their extraordinary capabilities of mounting rapid and massive type I IFN response to nu-cleic acids derived from virus,bacteria or dead cells.PDCs selectively express endosomal Toll-like receptor(TLR)7 and TLR9,which sense viral RNA and DNA re-spectively.Following type I IFN and cytokine responses,pDCs differentiate into antigen presenting cells and ac-quire the ability to regulate T cell-mediated adaptive immunity.The functions of pDCs have been implicated not only in antiviral innate immunity but also in immune tolerance,inflammation and tumor microenvironments.In this review,we will focus on TLR7/9 signaling and their regulation by pDC-specific receptors.