BACKGROUND AND PURPOSE:Our previous study reported that fructose intake increased systemic blood pressure and reduced nitric oxide (NO) in the nucleus tractus solitarius (NTS) due to oxidative stress and neuroinflammation. However, it remains unclear how reactive oxygen species (ROS) reduce NO and how this process impacts neuroinflammation in the NTS. This study aimed at investigating the effect of ROS on acetylation of high mobility group box 1 protein (HMGB1) in the NTS of fructose-induced hypertensive rats. EXPERIMENTAL APPROACH:Male Wistar-Kyoto (WKY) rats were fed with 10% fructose water to elevate blood pressure. Thereafter, CLI-095 and glycyrrhizic acid (GA) treatments were delivered for up to 2 weeks (1 mg·12 μL-1·day-1, by intracerebroventricular injection) to reduce the negative effects of toll-like receptor 4 (TLR4) and HMGB1 activation. KEY RESULTS:Two weeks of CLI-095 and GA treatment reduced systemic blood pressure and significantly preserved neuronal and endothelial nitric oxide synthase (nNOS and eNOS) availability against the inflammatory insults of fructose consumption. Both CLI-095 and GA halted the interaction of acetylated HMGB1 and TLR4. Two weeks of CLI-095 and GA treatment markedly reduced NTS inflammation (pro-inflammatory cytokines and microglial activation) and lowered serum norepinephrine levels. CONCLUSION AND IMPLICATIONS:Our data reveal novel pharmacological properties for CLI-095 and GA, which improved blood pressure and inflammatory conditions by decreasing the interaction of acetylated HMGB1 with TLR4. These findings challenge the commonly accepted dogma that essential hypertension is specifically mediated by neuroinflammation due to acetylated HMGB1 coupling to TLR4. LINKED ARTICLES:This article is part of a themed issue Recent Innovations in Targeting Redox Biology for Therapeutics. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v183.1/issuetoc.

Our previous study reported that the heterodimer of Angiotensin II Type I Receptor (AT1R) and Mu-Opioid Receptor 1 (MOR1) involves Nitric Oxide (NO) reduction which leads to elevation of blood pressure. Secondly, we showed that Toll-like Receptor 4 (TLR4) may be involved in the heterodimerization of AT1R and MOR1 in the brainstem Nucleus Tractus Solitarii (NTS), which regulates systemic blood pressure and gastric nitric oxide through the insulin pathway. Here, we investigated the role of microglial activation and TLR4 in the heterodimerization of AT1R and MOR1. Hypertensive rats were established after four weeks of fructose consumption. SBP of rats was measured using non-invasive blood pressure method. PLA technique was utilized to determine protein-protein interaction in the nucleus tractus solitarii. Results showed that the level of MOR-1 and AT1R was induced significantly in the fructose group compared with control. PLA signal potentially showed that AT1R and MOR1 were formed in the nucleus tractus solitarii after fructose consumption. Meanwhile, the innate immune cell in the CNS microglia was observed in the nucleus tractus solitarii using biomarkers and was activated. TLR4 inhibitor CLI-095, was administered to animals to suppress the neuroinflammation and microglial activation. CLI-095 treatment reduced the heterodimer formation of AT1R and MOR1 and restored nitric oxide production in the nucleus tractus solitarii. These findings imply that TLR4-primed neuroinflammation involves formation of heterodimers AT1R and MOR1 in the nucleus tractus solitarii which leads to increase in systemic blood pressure.

AIMS:Chronic hyperglycemia triggers overproduction of AKR1B1 (aldo-keto reductase family 1 member B) and receptor for advanced glycation end product (RAGE), which causes epithelial-mesenchymal transition (EMT) in the lens epithelial cells (LECs) of diabetic mellitus (DM) cataracts. However, it is unclear whether EMT in LECs is related to abnormal increase of SGLT2. Sodium glucose cotransporter 2 (SGLT2) inhibitor, also known as dapagliflozin (Dapa) can be used to treat diabetes. Here, we examined how Dapa or nano eye-drops (DapaN) reduce EMT in LECs of DM cataracts. The nano eye-drop provides an ophthalmic treatment that suppressed diabetic cataract progression and improved potency with reduced side effects. MAIN METHODS:SD rats were injected with streptozocin (STZ) (65 mg/kg, ip), nano-Dapa drops (0.456 mg/10 ml/eye) or Dapa (1.2 mg/kg/day) treatment for 6-12 weeks. Immunofluorescence staining was used for protein quantification of RAGE, SGLT2, N-cadherin and E-cadherin in the LECs of rats. KEY FINDINGS:In this study, Dapa applies nanotechnology-based delivery system and it contains polyvinylpyrrolidone (PVP) and HPBCD. Dapa showed therapeutic effect on DM cataracts, wherein it targeted EMT biomarker, E-cadherin. The nano-Dapa drops or oral Dapa inhibited SGLT2, suppressed AKR1B1 expression, decreased AcSOD2- and RAGE-induced EMT in diabetic cataracts. Our findings suggest that nanotechnology-based Dapa eye drops (Dapa-PVP-HPBCD) can effectively improve solubility of Dapa in aqueous solution. SIGNIFICANCE:Taken together, results suggest that the SGLT2-mediated DM cataract therapy may involve the AKR1B1-RAGE-AcSOD2-EMT pathway. The nano eye drops and Dapa show potential beneficial effects for cataract prevention. This study conveys new insights into cataract treatment and supplementation of nano-Dapa drops shows promising result in preventing diabetic cataracts.

Background Several studies have suggested mechanisms whereby excessive fructose intake increases blood pressure (BP). Glucose transporter 5 (GLUT5) is a fructose transporter expressed on enterocytes, and its involvement in the nucleus tractus solitarius (NTS)-modulated increase in BP following fructose intake remains unclear. Objectives Herein, we investigated whether NTS Glut5 knockdown (KD) can alleviate fructose-induced hypertension in rat models. Methods Male Wistar-Kyoto rats (6-8 weeks old; average weight: 230 g) were randomly assigned into 4 groups [control (Con), fructose (Fru), fructose + scrambled (Fru + S), and Fru + KD]. The Con group rats had ad libitum access to regular water, and the other 3 groups were provided 10% fructose water ad libitum for 4 weeks (2 weeks before lentiviral transfection in the Fru + S and Fru + KD groups). Glut5 short hairpin RNA was delivered into the NTS of rats using a lentivirus system. Fructose-induced hypertension was assessed via the tail-cuff technique, a noninvasive blood pressure measurement approach. GLUT5-associated and other insulin signaling pathways in the NTS of rats were assessed using immunofluorescence and immunoblotting analyses. We evaluated between-group differences using the Mann-Whitney U test or Kruskal-Wallis 1-way ANOVA. Results Compared with the Fru + S group, the Fru + KD group had reduced sympathetic nerve hyperactivity (48.8 +/- 3.2 bursts/min; P < 0.05), improved central insulin signaling, upregulated protein kinase B (AKT; 3.0-fold) and neuronal NO synthase (nNOS; 2.78-fold) expression, and lowered BP (17 +/- 1 mmHg, P < 0.05). Moreover, Glut5 KD restored signaling dependent on adenosine 5 '-monophosphate-activated protein kinase and reduced fructose-induced oxidative stress 2.0-fold, and thus decreased NAD(P)H oxidase in p67-phox 1.9-fold within the NTS. Conclusions Fructose-induced reactive oxygen species generates in the NTS of rats through GLUT5 and receptor for advanced glycation end products signaling, thus impairing the AKT-nNOS-NO signaling pathway and ultimately causing hypertension.

Chronic hyperglycemia triggers an abnormal rise in reactive oxygen species (ROS) that leads to blindness in patients with diabetes mellitus (DM) and cataracts. In this study, the effects of dapagliflozin, metformin and resveratrol on ROS production were investigated in lens epithelial cells (LECs) of animals with fructose-induced DM. LECs were isolated from patients without DM, or with DM devoid of diabetic retinopathy. Animals were treated with 10% fructose for 8 weeks to induce DM, which was verified by monitoring blood pressure and serum parameters. For drug treatments, 1.2 mg/day of dapagliflozin was given for 2 weeks, 500 mg/kg/day of metformin was given, and 10 mg/kg/day of resveratrol was given. Dihydroethidium was used to stain endogenous O2˙− production in vivo of the LECs. Superoxide production was expressed in the cataract of DM, or patients without DM. Sodium–glucose cotransporter 2 (SGLT2), glucose transporter 1 (GLUT1), GLUT5, the reduced form of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase subunit p47/p67-phox, NOX4 and RAGE were significantly increased in LECs with DM. In addition, the dapagliflozin treatment reduced GLUT5, p47/p67-phox, NADPH oxidase 4 (NOX4) and receptor for advanced glycation end products (RAGE) expressions. On the contrary, metformin or resveratrol inhibited p47-phox, GLUT5, and SGLT2 expressions, but not nuclear factor erythroid 2–related factor 2 (NRF2). In summary, dapagliflozin, metformin or resveratrol down-regulated p47-phox expression through SGLT2 inactivation and ROS reduction. These important findings imply that SGLT2 can be blocked to ameliorate oxidative stress in the cataracts of DM patients.

Opioids, a kind of peptide hormone involved in the development of hypertension, cause systemic and cerebral inflammation, and affects regions of the brain that are important for blood pressure (BP) control. A cause-and-effect relationship exists between hypertension and inflammation; however, the role of blood pressure in cerebral inflammation is not clear. Evidence showed that AT1R and μOR heterodimers’ formation in the NTS might lead to the progression of hypertension. In this study, we investigated the formation of the μOR/AT1R heterodimer, determined its correlation with μORs level in the NTS, and explored the role of TLR4-dependent inflammation in the development of hypertension. Results showed that Ang II increased superoxide and Iba-1 (microgliosis marker: ionized calcium-binding adaptor molecule (1) levels in the NTS of spontaneously hypertensive rats (SHRs). The AT1R II inhibitor, losartan, significantly decreased BP and abolished superoxide, Iba-1, TLR4 expression induced by Ang II. Furthermore, losartan significantly increased nNsOSS1416 phosphorylation. Administration of a μOR agonist or antagonist in the NTS of WKY and SHRs increased endogenous μ-opioids, triggered the formation of μOR/AT1R heterodimers and the TLR4-dependent inflammatory pathway, and attenuated the effect of depressor nitric oxide (NO). These results imply an important link between neurotoxicity and superoxides wherein abnormal increases in NTS endogenous μ-opioids promote the interaction between Ang II and μOR, the binding of Ang II to AT1R, and the activation of microglia. In addition, the interaction between Ang II and μOR enhanced the formation of the AT1R and μOR heterodimers, and inactivated nNOS-derived NO, leading to the development of progressive hypertension.

Studies demonstrated that the receptor of advanced glycation end products (RAGE) induced epithelial-mesenchymal transition (EMT) formation in the lens epithelial cells (LECs) of diabetic cataracts. This work investigated how 3H-1,2-dithiole-3-thione (D3T) reduces EMT formation in LECs of the fructose-induced diabetes mellitus (DM). LECs were isolated during cataract surgery from patients without DM or with DM. In a rat model, fructose (10% fructose, eight weeks) with or without D3T (10 mg/kg/day) treatment induced DM, as verified by blood pressure and serum parameter measurements. We observed that the formation of advanced glycation end products (AGEs) was significantly higher in epithelial human lens of DM (+) compared to DM (−) cataracts. Aldose reductase (AKR1B1), AcSOD2, and 3-NT were significantly enhanced in the rat lens epithelial sections of fructose-induced DM, however, the phosphorylation level of AMPKT172 showed a reversed result. Interestingly, administration of D3T reverses the fructose-induced effects in LECs. These results indicated that AMPKT172 may be required for reduced superoxide generation and the pathogenesis of diabetic cataract. Administration of D3T reverses the fructose-induced EMT formation the LECs of fructose-induced DM. These novel findings suggest that the D3T may be a candidate for the pharmacological prevention of cataracts in patients with DM.

The metabolic syndrome caused by excessive consumption of fructose is one of a variety of abnormal physiological phenomena, which is related to cardiovascular diseases. Our previous study reported that hypertension and central insulin resistance in WKY rats were caused by fructose intake. Several studies have been reported that the intake of fructose causes central inflammation. Toll‐like receptor 4 (TLR4) play a key role of innate immune signaling and has been implicated in fructose‐induced metabolic syndrome rats. We analyzed systolic blood pressure, biochemical parameter, and TLR4‐signaling pathway of WKY rats and fructose‐intake rats treated with vehicle or CLI‐095 (12 mg×d−1 for 14 days through osmotic mini‐pump). We found that the systolic blood pressure of rats treated CLI‐095 was improved to the normotensive group. The body weight change was recovered to the rats treated with the vehicle. Serum glucose treated with CLI‐095 was restored and stabilized compared with the fructose group. Immunoblotting results showed HMGB1 and TLR4 rather than TLR2, TLR7, TLR9 were induced in the brainstem nuclei area. MyD88, phosphorylated TGF beta‐activated kinase 1 (TAK1) and JNK were inhibited in the CLI‐095 group. Neuroinflammatory indicator iNOS was reduced in the therapy group. These data demonstrated that TLR4 in the central brain participates in the regulation of blood pressure, glucose, and body weight, which suggested HMGB1‐TLR4 might be a potential target for therapeutic interventions in the condition with metabolic syndrome combined with neuroinflammation.Support or Funding InformationThis work was supported by the Ministry of Science and Technology grants 107‐2320‐B‐075B‐001 (to P.W.C.) and 107‐2320‐B‐075B‐002‐MY2 (to C.J.T.) and Kaohsiung Veterans General Hospital grants VGHKS108‐162 (to Y.T.L.) and VGHUST108‐G3‐1‐3 (to Y.T.L.).

Background Inflammation is a common pathophysiological trait found in both hypertension and cardiac vascular disease. Recent evidence indicates that fractalkine (FKN) and its receptor CX3CR1 have been linked to inflammatory response in the brain of hypertensive animal models. Here, we investigated the role of CX3CR1-microglia in nitric oxide (NO) generation during chronic inflammation and systemic blood pressure recovery in the nucleus tractus solitarii (NTS). Methods The hypertensive rat model was used to study the role of CX3CR1-microglia in NTS inflammation following hypertension induction by oral administration of 10% fructose water. The systolic blood pressure was measured by tail-cuff method of non-invasive blood pressure. The CX3CR1 inhibitor AZD8797 was administered intracerebroventricularly (ICV) in the fructose-induced hypertensive rat. Using immunoblotting, we studied the nitric oxide synthase signaling pathway, NO concentration, and the levels of FKN and CX3CR1, and pro-inflammatory cytokines were analyzed by immunohistochemistry staining. Results The level of pro-inflammatory cytokines IL-1β, IL-6, TNF-α, FKN, and CX3CR1 were elevated two weeks after fructose feeding. AZD8797 inhibited CX3CR1-microglia, which improved the regulation of systemic blood pressure and NO generation in the NTS. We also found that IL-1β, IL-6, and TNF-α levels were recovered by AZD8797 addition. Conclusion We conclude that CX3CR1-microglia represses the nNOS signaling pathway and promotes chronic inflammation in fructose-induced hypertension. Collectively, our results reveal the role of chemokines such as IL-1β, IL-6, and TNF-α in NTS neuroinflammation with the involvement of FKN and CX3CR1.

Purpose: Cataracts in patients with diabetes mellitus (DM) are a major cause of blindness in developed and developing countries. This study aims to examine whether the generation of reactive oxygen species (ROS) via the increased expression of glucose transporters (GLUTs) and the receptor for advanced glycation end products (RAGE) influences the cataract development in DM. Methods: Lens epithelial cells (LECs) were isolated during cataract surgery from patients without DM or with DM, but without diabetic retinopathy. In a rat model, fructose (10% fructose, 8 or 12 weeks) with or without dapagliflozin (1.2 mg/day, 2 weeks) treatment did induce DM, as verified by blood pressure and serum parameter measurements. Immunofluorescence stainings and immunoblottings were used to quantify the protein levels. Endogenous O2˙¯ production in the LECs was determined in vivo with dihydroethidium stainings. Results: We investigated that GLUT levels in LECs differed significantly, thus leading to the direct enhancement of RAGE-associated superoxide generation in DM patients with cataracts. Superoxide production was significantly higher in LECs from rats with fructose-induced type 2 DM, whereas treatment with the sodium-glucose cotransporter 2 (SGLT2) inhibitor dapagliflozin prevented this effect in fructose-fed rats. Protein expression levels of the sodium/glucose cotransporter 2 (SGLT2), GLUT1, GLUT5, the nicotinamide adenine dinucleotide phosphate reduced form (NADPH) oxidase subunit p67-phox, NOX2/4 and RAGE were upregulated in fructose-fed animals, whereas dapagliflozin treatment reversed these effects. Conclusions: In rats with fructose-induced DM, dapagliflozin downregulates RAGE-induced NADPH oxidase expression in LECs via the inactivation of GLUTs and a reduction in ROS generation. These novel findings suggest that the SGLT2 inhibitor dapagliflozin may be a candidate for the pharmacological prevention of cataracts in patients with DM.

Essential hypertension is the most common cardiovascular disease and is a major risk factor for stroke and heart disease. It is known the nucleus tractus solitarii (NTS) located on the dorsal medulla play an important role in regulating cardiovascular responses. More recently, metabolic syndrome‐induced phosphorylated AMPK defect has been demonstrated to underline the initiation of superoxide elevation. In recent years, fructose induces a decrease in SOD2 activity, which leads to a large increase in superoxide and a decrease in the pathway of Akt‐nNOS. Studies have found that stimulation with abnormal diet factor can promote the reduction of Sirt3 and increase the content of superoxide. Therefore, we hypothesize that the SOD2 activity in the NTS was inhibited via Sirt3‐AMPK dysfunction induced superoxide expression and hypertension. During this study, the rats were fed 10% fructose with or without natural compound 3H‐1,2‐Dithiole‐3‐thione (D3T) for 4 weeks with the D3T treatment beginning at week 2. The systolic blood pressure and serum fasting high‐density lipoprotein decreased significantly in the D3T‐treated group compared fructose‐fed group after feeding D3T for 2 weeks. The protein levels of Sirt3, and AMPKT172 showed a significant increase in the D3T‐treated group compared fructose group after feeding D3T for 2 weeks by immunoblotting analyses. However, the acetylation of SOD2 were significantly decreased in the D3T‐fed group compared the fructose‐fed groups. Superoxide anion expression levels in the NTS were clearly eliminated in the D3T‐fed group compared the fructose‐fed groups. Collectively, fructose induced Sirt3 and AMPK defect and acetylation of SOD2 in the NTS of fructose‐fed rats, while treatment of D3T in the fructose‐fed rat reversed the effect. This study suggest that fructose‐induced hypertension may through Sirt3‐AMPK to mediate the acetylation of SOD2 and causes superoxide generation in the central nervous system. Our findings suggested a better understanding of complex cardiovascular neural pathways and may provide a new link between hypertension and neuronal superoxide that could be targeted for the treatment of cardiometabolic diseases.Support or Funding InformationThis work was supported by funding from the Ministry of Science and Technology (MOST‐107‐2320‐B‐075B‐002).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

SCOPEIn the Natural Medicines database, coenzyme Q10 (CoQ10) is classified as possibly effective for the treatment of hypertension. Patients with hypertension frequently have a significant deficiency of the antioxidant CoQ10. Furthermore, reactive oxygen species are overproduced in the nucleus tractus solitarii (NTS) during the cardiovascular regulation of hypertension in vivo. However, the molecular mechanisms by which CoQ10 modulates cardiovascular functions in the NTS are unclear. In this study, the effects of CoQ10 on superoxide generation, downstream NO signaling in the NTS, and blood pressure were evaluated in rats with fructose-induced hypertension.METHODS AND RESULTSTreatment with oral CoQ10 for 4 weeks abolished nicotinamide adenine dinucleotide phosphate-oxidase (NADPH oxidase) activation, decreased p38 phosphorylation, and increased superoxide dismutase 2 production in the NTS of fructose-fed rats. The serum levels of uric acid decrease in response to CoQ10 treatment in fructose-fed rats. Oral CoQ10 reduced blood pressure by inducing Akt and nNOS phosphorylation in NTS of fructose-induced hypertensive rats.CONCLUSIONOral CoQ10 decreases blood pressure by negatively regulating fructose-induced NADPH oxidase levels, abolishing ROS generation, reducing p38 phosphorylation, and enhancing the Akt-nNOS pathway in the NTS. These results support the beneficial effects of CoQ10 in oxidative stressassociated hypertension.

Recent studies have indicated that several anti-hypertensive drugs may delay the development and progression of Alzheimer's disease (AD). However, the relationships among AD, hypertension, and oxidative stress remain to be elucidated. Here, we aimed to determine whether reactive oxygen species (ROS) reduction by resveratrol in the brain leads to cognitive impairment reduction in rats with angiotensin II (Ang-II)-induced early AD. Male Wistar Kyoto (WKY) rats with Ang-II-induced AD were treated with losartan or resveratrol for two weeks. Our results show decreased blood pressure, increased hippocampal brain-derived neurotrophic factor (BDNF) level, and decreased nucleus tractus solitarius (NTS) ROS production in the Ang-II groups with losartan (10 mg/kg), or resveratrol (10 mg/kg/day) treatment. Furthermore, losartan inhibition of hippocampal TauT231 phosphorylation activated AktS473 phosphorylation, and significantly abolished Ang-II-induced Aβ precursors, active caspase 3, and glycogen synthase kinase 3β (GSK-3β)Y216 expressions. Consistently, resveratrol showed similar effects compared to losartan. Both losartan and resveratrol restored hippocampal-dependent contextual memory by NADPH oxidase 2 (NOX2) deletion and superoxide dismutase 2 (SOD2) elevation. Our results suggest that both losartan and resveratrol exert neuroprotective effects against memory impairment and hippocampal damage by oxidative stress reduction in early stage AD rat model. These novel findings indicate that resveratrol may represent a pharmacological option similar to losartan for patients with hypertension at risk of AD during old age.

Hypertension is a risk factor for several cardiovascular disease, such as stroke, myocardial infarction, and heart failure. Our previous studies demonstrated that fructose inhibit NO release through P-AktS473-P-nNOSS1416 pathway in the nucleus tractus solitarii (NTS), leading to increased blood pressure (BP). Evidence suggests that long-term intake of fructose increase the sterile inflammatory response, producing mRNA level of proinflammatory cytokines (e.g. IL-1beta, IL-6, TNF-alpha) in central nervous system (CNS). Furthermore, neuroinflammation is known to be associated with hypertension; however, the mechanism is still unclear. Therefore, this study is to determine whether NO release will be inhibited through neuroinflammatory response in the CNS of fructose-induced hypertension. The hypertensive group were fed with 10% fructose for 1, 2, or 4 weeks respectively. Systolic blood pressure was measured by tail-cutoff method. Immunoblotting or immunohistochemistry analyses were used to quantify protein expression levels of inflammatory cytokines or signal pathway. After that, we treated fructose-induced hypertensive rat by intracerebroventricular infusion of AZD8797 (CX3CR1 antagonist) via osmotic minipump for 2 weeks. In our results, we found the IL-1beta, IL-6, CX3CL1 and CX3CR1 have high expression levels in the NTS and blood pressure (BP) increased significantly after fructose feeding for 2 weeks. However, the BP restores to normal level after administration of AZD8797 groups. Besides, the protein level of P-AktS473 and P-nNOSS1416 showed a significant recovery in the AZD8797 group compared to the fructose-vehicle group. These findings suggest that fructose induced hypertension may through CX3CL1-CX3CR1 caused neuroinflammation which induced P-AktS473-P-nNOSS1416 pathway defect in the central neuronal system. Based on our findings, there is a novel link between hypertension and neuronal inflammation that could be targeted for treatment of cardiometabolic diseases.

Abstract MicroRNAs (miRNAs) are small non-coding RNAs, approximately 20-23 nucleotides in length. The specific functions of miRNAs involved in several cellular processes, including metabolism, development, differentiation, proliferation, apoptosis and tumorigenesis. In order to identify the invasion/metastasis related miRNA in lung cancer, the cell line model (lowly invasive lung cancer cells, CL1-0, and highly invasive lung cancer cells, CL1-5) and human miRNA microarray were carried out. In this study, we selected one of the invasion/metastasis related miRNAs, miR-127, derived from our microarray results for further functional characterization. Recently reports have indicated that human miR-127 was highly expressed in normal prostate and bladder tissues but was remarkably down-regulated in the corresponding tumors. miR-127 may function as a tumor suppressor in human bladder and prostate cancer cells. However, the role of the miR-127 in lung cancer cell proliferation, migration and invasion ability still remains unclear. The miR-127 expression level in various invasive capability lung cancer cell lines were also verified using stem-loop real time RT-PCR. We have constructed and established a miR-127 constitutive expression system in CL1-0 lung cancer cells using pSilencer3.1 vector, and we also knockdown the miR-127 expression in CL1-5 and A549 lung cancer cells using anti-miR127 modified oligonucleotide. Our results shown that over-expression of miR-127 can increase lung cancer cell proliferation, migration, and invasion ability. Finally, we suggest that miR-127 may be acting like an oncogene in lung adenocarcinoma cells. These results will provide important information and help us to understand the roles of miR127 in lung cancer cells. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 3947. doi:10.1158/1538-7445.AM2011-3947