Expression Levels Of Caveolin-1 Research Articles

P2.01-48 Enhanced Expression of miR-204 Reduces Cisplatin Resistance in NSCLC

Non-small cell lung cancer (NSCLC) is the most common and lethal human malignant tumor worldwide. Platinum-based chemotherapy is still the mainstay treatment for NSCLC. However, long-term chemotherapy usually induces serious drug resistance. Accordingly, it is valuable to develop treatment strategies that could reduce the resistance against platinum-based drugs in NSCLC cells. Cisplatin-resistant NSCLC models in A549 and PC9 cell lines (CR-A549 and CR-PC9) were established through long-term exposure to cisplatin. Expression levels of caveolin-1 (CAV-1) and miR-204 in A549, PC9, CR-A549, and CR-PC9 cell lines were detected by western blot and q-PCR, respectively. Regulation of CAV-1 by miR-204 was assessed by bioinformatic analysis, western blot and luciferase reporter assay. The role of the miR-204/CAV-1 axis in regulating cisplatin resistance in NSCLC cells was tested by MTT assay. Interactions with Bcl-2-associated agonist of cell apoptosis (Bad) and Bcl-xl (Bcl-2) were evaluated by co-immunoprecipitation assay. Flow cytometry was used to detect the apoptosis of NSCLC cell lines. We observed significant upregulation of CAV-1 expression and a significant decrease of miR-204 expression in CR-A549 and CR-PC9 cells compared to their parental A549 and PC9 cells. Furthermore, we detected down regulation of miR cells. We then found that enforced expression of miR-204 can resensitize CR-A549 and CRPC9 cells to cisplatin treatment through the inhibition of CAV-1 expression. Thus, we declared that recovery of miR-204 expression was able to sensitize the cisplatin-induced mitochondrial apoptosis observed in CR-A549 and CR-PC9 cells through suppression of the caveolin-1/AKT/Bad pathway. This study suggested that enhanced expression of miR-204 could increase the sensitivity of cisplatin treatment by suppression of caveolin-1/AKT/Bad pathway, which concluded that overexpression of miR-204 could be a potential approach for enhancing the sensitivity in NSCLC cells with cisplatin resistance.

Dipeptidyl peptidase-4 plays a pathogenic role in BSA-induced kidney injury in diabetic mice

Diabetic kidney disease (DKD) is appeared to be higher risk of declining kidney function compared to non-diabetic kidney disease with same magnitude of albuminuria. Epithelial-mesenchymal transition (EMT) program of tubular epithelial cells (TECs) could be important for the production of the extracellular matrix in the kidney. Caveolin-1 (CAV1), dipeptidyl peptidase-4 (DPP-4) and integrin β1 have shown to be involved in EMT program. Here, we found diabetic kidney is prone for albuminuria-induced TECs damage and DPP-4 plays a vital role in such parenchymal damages in diabetic mice. The bovine serum albumin (BSA) injection induced severe TECs damage and altered expression levels of DPP-4, integrin β1, CAV1, and EMT programs including relevant microRNAs in type 1 diabetic CD-1 mice when compared to non-diabetic mice; teneligliptin (TENE) ameliorated these alterations. TENE suppressed the close proximity among DPP-4, integrin β1 and CAV1 in a culture of HK-2 cells. These findings suggest that DPP-4 inhibition can be relevant for combating proteinuric DKD by targeting the EMT program induced by the crosstalk among DPP-4, integrin β1 and CAV1.

Subanesthetic Dose of Ketamine Improved CFA-induced Inflammatory Pain and Depression-like Behaviors Via Caveolin-1 in Mice.

Ketamine, a commonly used nonbarbiturate anesthetic drug, possesses antidepressant properties at subanesthetic doses; however, the underlying mechanisms remain unclear. The analgesic and antidepressant effects of ketamine were explored using a complete Freund adjuvant (CFA)-induced peripheral inflammatory pain model in vivo. Mice were first divided into sham or CFA injection group randomly, and were observed for mechanical hyperalgesia, depression-like behavior, and mRNA expression of caveolin-1. Then ketamine was administered in CFA-treated mice at day 7. The behavioral testing results revealed mechanical hyperalgesia and depression in mice from days 7 to 21 after CFA injection. Ketamine reversed depression-like behaviors induced by CFA injection. It also restored the brain-regional expression levels of caveolin-1 in CFA-treated mice. In addition, caveolin-1 mRNA and protein expression were increased in the prefrontal cortex and nucleus accumbens of CFA-treated mice. However, ketamine reversed the increase in caveolin-1 expression in the ipsilateral and contralateral prefrontal cortex and nucleus accumbens, supporting the distinct roles of specific brain regions in the regulation of pain and depression-like behaviors. In CFA-treated mice that exhibited pain behavior and depression-like behavior, ketamine reversed depression-like behavior. The prefrontal cortex and nucleus accumbens are the important brain regions in this regulation network. Despite these findings, other molecules and their mechanisms in the signal pathway, as well as other regions of the brain in the pain matrix, require further exploration.

Clinical implications of hypoxia-inducible factor-1α and caveolin-1 overexpression in isocitrate dehydrogenase-wild type glioblastoma multiforme.

Glioblastoma multiforme (GBM) is the most common type of primary brain tumour in adults, and presents a very low survival rate. Isocitrate dehydrogenase (IDH)1/2 mutations have been found in ~12% of glioblastomas and are associated with long-term GBM survival. However, the risk factors that influence the prognosis of IDH-wild type GBM remain unclear. Hypoxia-inducible factor (HIF)-1α, an important oxygen-regulated transcription factor, has been demonstrated to serve a crucial role in tumour development and to be associated with a poor prognosis. In addition, caveolin-1 (CAV1) is a plasma membrane organizing protein, the expression of which can also be regulated by a hypoxic microenvironment. The present study therefore aimed to examine the expression levels of HIF-1α and CAV1, and their association with GBM prognosis. Reverse transcription-quantitative polymerase chain reaction and western blotting were performed to analyse the expression levels of HIF-1α and CAV1 in paired GBM tumour and adjacent non-tumour tissues. Immunohistochemistry was used to analyse the expression of the two proteins in paraffin-embedded tissues obtained from 42 patients with IDH-wild type GBM. Statistical analyses were performed to examine the correlation between HIF-1α and CAV1 expression and patient prognosis. The results revealed hat the expression levels of HIF-1α and CAV1 were upregulated in IDH-wild type GBM tissues compared to their paired non-tumour tissues (P<0.001). The expression of CAV1 was significantly correlated with high HIF-1α expression (P<0.01). In addition, overexpression of HIF-1α and CAV1 was markedly associated with a poor prognosis (P<0.001). In conclusion, HIF-1α and CAV1 may represent potential biomarkers for IDH-wild type GBM prognosis and potential targets for the development of therapies extending GBM survival.

Resveratrol blocks atherosclerosis development by inhibiting IL-1β in macrophages induced by cholesterol

Resveratrol is a polyphenolic compound that exhibits antiinflammatory and cardioprotective properties. In this study we investigated the protective role of resveratrol on the inflammatory activation of macrophages during pathogenesis of atherosclerosis. Macrophage Ana-1 cells were stimulated by cholesterol and resveratrol, and the cell culture supernatant was collected to treat human umbilical vein endothelial cells (HUVECs). The release of IL-1? into the Ana-1 cell supernatant was quantified by ELISA. Expression of the adhesion molecule ICAM-1 and E-selectin in HUVECs were examined by Western-blotting. Additionally, the adhesion of monocytes in HUVECs under different conditions was tested by cell adhesion analyses. The results indicated that the high cholesterol treatment increased the expression level of IL-1?, while pretreatment with resveratrol inhibited this induction of IL-1? in Ana-1 cells. Resveratrol inhibited the adhesion of monocytes to the endothelium at least partly through inhibition of IL-1? expression in macrophages. Moreover, the expression level of caveolin-1 significantly increased after the pretreatment with resveratrol, indicating that resveratrol enhances reverse cholesterol transport (RCT) in macrophages. Our study indicated that resveratrol has significant antiinflammatory effects and can be considered as a candidate molecule to prevent atherosclerosis.

The effect of caveolin-1 knockdown on interleukin-1β-induced chemokine (C-C motif) ligand 2 expression in synovial fluid-derived fibroblast-like synoviocytes from patients with rheumatoid arthritis.

Rheumatoid arthritis (RA) is a chronic autoimmune disease leading to destructive changes in peripheral joints and their irreversible deformity. The influx of chemoattractant-mediated inflammatory cells to the joints is one of the main features of RA. The aim of this study was to investigate the effect of a knockdown of caveolin-1 (CAV1), a known regulator of multiple cell signaling pathways, on chemokine (C-C motif) ligand 2/monocyte chemoattractant protein-1 (CCL2/MCP-1) expression in synovial fluid-derived fibroblast-like synoviocytes (sfd-FLSs) obtained from patients with RA. Primary cell cultures of sfd-FLSs were established from RA synovial fluids. Cells were transiently transfected with small interfering RNA (siRNA) specific for CAV1, and then incubated with interleukin (IL)-1β to induce CCL2 expression. The expression levels of CAV1 and CCL2 were assessed at transcript level, using quantitative polymerase chain reaction (qPCR) and at protein level by enzyme-linked immunosorbent assay (ELISA) and western blotting analysis. A transient CAV1 knockdown in sfd-FLSs resulted in a decrease in the IL-1β-induced CCL2 mRNA expression level vs non-transfected cells and cells transfected with non-targeting siRNA. The concentration of secreted CCL2 was not affected significantly. Our study demonstrates that CCL2 expression in sfd-FLSs is CAV1-dependent, but only at transcript level. As the function of CAV1 has not been unequivocally determined, more studies are needed to confirm the role of CAV1 in inflammatory processes related to RA.

Eps15 homology domain 1 promotes the evolution of papillary thyroid cancer by regulating endocytotic recycling of epidermal growth factor receptor.

Papillary thyroid cancer (PTC) is the most common type of thyroid malignancy, and it is often observed to overexpress epidermal growth factor receptor (EGFR). Previous research has indicated that EH domain-containing 1 (EHD1) is associated with EGFR-mediated endocytotic recycling in multiple tumor types. The objective of the present study was to determine the protein expression levels and clinical significance of EHD1, EGFR, caveolin-1 (CAV-1) and RAB11 family interacting protein 3 (RAB11FIP3) in PTC. PTC specimens were analyzed for EHD1, EGFR, CAV-1 and RAB11FIP3 expression via immunohistochemistry and western blotting. The associations between protein expression and clinicopathological features were assessed. EHD1, EGFR, CAV-1 and RAB11FIP3 expression levels were increased in human PTC. Additionally, the expression level of EHD1 protein was significantly associated with tumor size, lymph node metastasis and EGFR expression (P<0.05). CAV-1 was associated with tumor size and EGFR expression (P<0.05). EGFR was only associated with lymph node metastasis (P=0.027) and RAB11FIP3 was not associated with any clinicopathological characteristics. The correlations between EHD1 and EGFR (r=0.564, P<0.05), CAV-1 (r=0.865, P<0.01) and RAB11FIP3 (r=0.504, P<0.05) were statistically significant. Overall, EHD1, CAV-1 and RAB11FIP3, which are key proteins in endocytotic recycling, promote PTC tumorigenesis through the regulation of the transport of EGFR.

The effects of caveolin1 on β cell proliferation.

Our study aims to access the influence of caveolin1 (CAV1) on β cell expression profiles. We knocked down the expression of CAV1 in both NIT-1 cells and islets isolated from C57BL/6J mice using an RNA interference technique, which was realized by the transfer of an shRNA vector targeting CAV1 mRNA into NIT-1 cells or islets through latent virus infection. First, we identified the change in gene expression profiles in islets, in which the CAV1 expression level was down-regulated, as ascertained by mouse gene expression microarray, and the results showed that pathways related to β cell proliferation and pancreatic secretion functions were significantly influenced. The results of MTT demonstrated that the knockdown of CAV1 expression in NIT-1 cells promoted proliferation. The protein array results showed that pro-apoptotic cytokines were down-regulated in the NIT-1 cell line with CAV1 knockdown. These findings suggest that CAV1 might be involved in apoptosis and proliferation regulation in β cells, and therefore could be a potential target for the development of novel therapies for diabetes mellitus.

Effects of levothyroxine on pregnancy outcomes and caveolin-1 expression in rat models of thyroid dysfunction

To investigate the effect of thyroid hormone levothyroxine (L-T4 ) on pregnancy outcomes and expression of caveolin-1 in rat models of thyroid dysfunction. Materials and Methods: One hundred Wistar rats (SPE grade, 60 female and 40 male) were randomly divided into three groups of control, model (hypothyroidism), and treatment (hypothyroidism + therapy with L-T4 ) groups. Pregnant rats were investigated for pregnancy outcomes (litter size, number of dead fetus, placental index, body weight and length, and tail length of neonatal rat). Expression of caveolin-1 in placenta was analyzed using qRT- PCR and Western blot. Results: The modeling success rate was 75%. Investigations showed that the litter size, number of dead fetus, placental index, body and length, and tail length of neonatal rat in the models were significantly lower than those in control rats (p p p > 0.05). The expression level of caveolin-1 in placenta was significantly higher in models than that in control (p Conclusion: Propylthiouracil (PTU) can successfully induce hypothyroidism in rat and hypothyroidism can seriously hinder the pregnancy process, resulting in developmental disorders in fetal rat.

Effect of Andrographolide on Gene Expression Profile and Intracellular Calcium in Primary Rat Myocardium Microvascular Endothelial Cells.

Andrographolide (ANDRO) is a diterpene lactone compound with extensive biological effects, such as antibacterial, antitumor and treatment of cardiovascular diseases. Until now, studies on the pharmacological functions of ANDRO are still in progress. However, little is known about the gene expression profile and calcium response of endothelial cells to ANDRO. In this study, we used a microarray technology to investigate the gene expression responses in primary rat myocardium microvascular endothelial cells treated with 10 μg/mL ANDRO. The expression of caveolin-1 and 1-phosphatidylinositol 4, 5-bisphosphate phosphodiesterase δ3 was verified by RT-PCR and western blot. In addition, we investigated the effect of ANDRO on intracellular calcium induced by exogenous adenosine triphosphate and on inflammatory response induced by lipopolysaccharide. Results showed that ANDRO treatment induced an abundance of differential expressed genes, exhibiting a multitarget regulatory effect. ANDRO significantly decreased caveolin-1 and phosphodiesterase δ3 expression, lipopolysaccharide-induced IL-6 and TNF-α levels and expression of several chemokine genes, which are associated with reducing inflammation response and decreasing calcium release without affecting normal endothelia cell function, suggesting that ANDRO may be a potential candidate to treat cardiovascular diseases with less toxicity.

Effects of combined helium pre/post-conditioning on the brain and heart in a rat resuscitation model.

The noble gas helium induces cardio- and neuroprotection by pre- and post-conditioning. We investigated the effects of helium pre- and post-conditioning on the brain and heart in a rat resuscitation model. After approval by the Animal Care Committee, 96 Wistar rats underwent cardiac arrest for 6min induced by ventricular fibrillation. Animals received 70% helium and 30% oxygen for 5min before cardiac arrest and for 30min after restoration of spontaneous circulation (ROSC). Control animals received 70% nitrogen and 30% oxygen. Hearts and brains were excised after 2, 4h or 7days. Neurological degeneration was evaluated using TUNEL and Nissl staining in the hippocampal CA-1 sector. Cognitive function after 7days was detected with the tape removal test. Molecular targets were measured by infrared western blot. Data are shown as median [Interquartile range]. Helium treatment resulted in significantly less apoptosis (TUNEL positive cells/100 pixel 73.5 [60.3-78.6] vs.78.2 [70.4-92.9] P=0.023). Changes in Caveolin-3 expression in the membrane fraction and Hexokinase-II in the mitochondrial fraction were observed in the heart. Caveolin-1 expression of treated animals significantly differed from control animals in the membrane fraction of the heart and brain after ROSC. Treatment with helium reduced apoptosis in our resuscitation model. Differential expression levels of Caveolin-1, Caveolin-3 and Hexokinase II in the heart were found after helium pre- and post-conditioning. No beneficial effects were seen on neurofunctional outcome.

The level of caveolin-1 expression determines response to TGF-\u03b2 as a tumour suppressor in hepatocellular carcinoma cells

Hepatocellular carcinoma (HCC) is a heterogeneous tumour associated with poor prognostic outcome. Caveolin-1 (CAV1), a membrane protein involved in the formation of caveolae, is frequently overexpressed in HCC. Transforming growth factor-beta (TGF-β) is a pleiotropic cytokine having a dual role in hepatocarcinogenesis: inducer of apoptosis at early phases, but pro-tumourigenic once cells acquire mechanisms to overcome its suppressor effects. Apoptosis induced by TGF-β is mediated by upregulation of the NADPH oxidase NOX4, but counteracted by transactivation of the epidermal growth factor receptor (EGFR) pathway. Previous data suggested that CAV1 is required for the anti-apoptotic signals triggered by TGF-β in hepatocytes. Whether this mechanism is relevant in hepatocarcinogenesis has not been explored yet. Here we analysed the TGF-β response in HCC cell lines that express different levels of CAV1. Accordingly, stable CAV1 knockdown or overexpressing cell lines were generated. We demonstrate that CAV1 is protecting HCC cells from TGF-β-induced apoptosis, which attenuates its suppressive effect on clonogenic growth and increases its effects on cell migration. Downregulation of CAV1 in HLE cells promotes TGF-β-mediated induction of the pro-apoptotic BMF, which correlates with upregulation of NOX4, whereas CAV1 overexpression in Huh7 cells shows the opposite effect. CAV1 silenced HLE cells show attenuation in TGF-β-induced EGFR transactivation and activation of the PI3K/AKT pathway. On the contrary, Huh7 cells, which do not respond to TGF-β activating the EGFR pathway, acquire the capacity to do so when CAV1 is overexpressed. Analyses in samples from HCC patients revealed that tumour tissues presented higher expression levels of CAV1 compared with surrounding non-tumoural areas. Furthermore, a significant positive correlation among the expression of CAV1 and TGFB1 was observed. We conclude that CAV1 has an essential role in switching the response to TGF-β from cytostatic to tumourigenic, which could have clinical meaning in patient stratification.

Missing-in-metastasis B (MIM-B) combined with caveolin-1 promotes metastasis of hepatocellular carcinoma.

BackgroundIncreasing amounts of evidence indicate that Missing in metastasis B (MIM-B) promotes cancer metastasis. Here, we sought to better understand the mechanism through which MIM-B promotes tumor metastasis in hepatocellular carcinoma (HCC).MethodsWe performed confocal microscopy analysis to determine the distributions of MIM-B and caveolin-1 and conducted co-immunoprecipitation assays to detect the interactions between MIM-B and caveolin-1 in vitro. We performed transwell assays to analyze the invasive ability of HCC cells. Changes in the expression levels of key genes and some molecular makers were detected by immunohistochemistry and western blotting in HCC tissue samples.ResultsWe found that MIM-B co-localizes with caveolin-1 and demonstrated that MIM-B and caveolin-1 interact in vitro. Repressing MIM-B and caveolin-1 expression inhibited the epidermal growth factor receptor signaling pathway. We overexpressed MIM-B and caveolin-1 in Hep3B cells, which enhanced Hep3B cell invasiveness. Furthermore, MHCC97H cell invasiveness was significantly decreased in cells in which MIM-B and caveolin-1 expression was inhibited. Additionally, we found that MIM-B and caveolin-1 were expressed at higher levels in HCC tissues than in paired normal tissues. Moreover, HCC patients with MIM-B and caveolin-1 up-regulation experienced significantly worse outcomes than controls (P < 0.001), and HCC patients with high MIM-B and caveolin-1 expression levels often developed pulmonary metastasis (P < 0.001).ConclusionsMIM-B combined with caveolin-1 promotes metastasis of HCC, and elevated MIM-B and caveolin-1 expression levels are associated with a poor prognosis in HCC patients; therefore, MIM-B and caveolin-1 may represent novel targets for the diagnosis and treatment of HCC.

337 Downregulated Caveolin-1 expression in monocytes may contribute to the pathogenesis of psoriasis

Caveolin-1 (CAV-1) is a membrane protein that is essential for the formation of flask-shaped membrane invaginations known as caveolae. CAV-1 regulates a variety of signaling molecules and receptors, and aberrant CAV-1 expression is involved in a variety of diseases. Our previous study showed aberrant reduction of CAV-1 in the epidermis of psoriasis patient, which leads to enhance phosphorylation of STAT3 and cytokine production, suggesting that decreased CAV-1 may contribute to the psoriatic inflammation. Not only keratinocytes, immune cells are major players in psoriasis. Migrated immune cells produce variety of effector cytokines to affect keratinocytes proliferation and induce psoriatic inflammation. The purpose of this study is to determine whether CAV-1 is involved in the leucocytes function of psoriasis and investigate their role in the pathogenesis of psoriasis. We first determined CAV-1 expression levels of peripheral blood mononuclear cells (PBMCs) and polymorphonuclear cells (PMNs) isolated from patients with psoriasis and control subjects by quantitative PCR and immunoblotting. Immunocytochemistry and magnetic cell isolation were further conducted to identify which cells were aberrant on CAV-1 expression in PBMCs. As a result, mRNA and protein levels of CAV-1 in PBMCs and PMNs were significantly decreased in psoriasis patients compared to that in control subjects. In PBMCs, reduction of CAV-1 was observed significantly in CD14 positive monocytes. We further assessed function of CAV-1 decreased monocytes on cytokine production and their migration. RNA interference of CAV-1 in isolated monocytes represented elevated levels of IL-1b and IL-6 under LPS stimulation compared to controlled monocytes. Furthermore, migration assay revealed that CAV-1 silenced monocytes had enhanced chemotaxis activity induced by MCP-1. Our results suggest that aberrant CAV-1 expression on monocytes may be involved in the pathogenesis of psoriasis mediating cell migration and cytokine production.

Caveolin-1 expression as a prognostic marker in triple negative breast cancers of Asian women

BackgroundTriple-negative breast cancers (TNBCs) are defined by their lack of oestrogen receptor, progesterone receptor and epidermal growth factor receptor 2. Although heterogeneous, the majority are aggressive and treatment options are...

Abstract 1770: The association between sarcopenia and cellular senescence of cancer associated fibroblast in pancreatic cancer

Abstract Background: Sarcopenia is a syndrome that is characterized by progressive and generalized loss of skeletal muscle mass and strength. Several recent studies have shown that sarcopenia has prognostic significance for patients with malignant disease, including cancers of the esophagus, colon, pancreas and liver. On the other hand, cellular senescence is gaining increased attention from clinicians and researchers, yet incompletely understood role in the development of malignant disease. Previous studies reported that Caveolin-1 expression in cancer associated fibroblast is associated with cancer progression, and it is known in recent years that caveolin-1 plays a major role in controlling cellular senescence. Therefore, we hypothesized that sarcopenia causes cellular senescence of cancer associated fibroblast (CAF). The aim of this study is to clarify the between sarcopenia and cellular senescence of CAF through Caveolin-1 in pancreatic cancer. Methods: All consecutive patients with pancreatic cancer underwent curative resection between January 2004 and December 2014 were enrolled in this retrospective study. Skeletal muscle and visceral fat amount at the third lumbar vertebra (L3) in the inferior direction were quantified using enhanced computed tomography. The patients were divided into two groups, with and without sarcopenia, based on Japan Society of Hepatology guidelines for sarcopenia. Next, Caveolin-1 expression was analyzed by immunohistochemistry (IHC). Moreover, we investigated the relationship between sarcopenia and Caveolin-1 expression. Results: Forty-one (31.5%) of 130 patients were identified with sarcopenia. Kaplan-Meier analysis showed that patients with sarcopenia had a significantly worse cancer specific survival than patients without sarcopenia (3-year-survival: 37.2% vs 50.4%, respectively, p = 0.038). While, a high level of Caveolin-1 expression was detected in 29.2% (38/130) of our samples. High Caveolin-1 expression in CAF significantly correlated with poor prognoses with respect to overall survival (log-rank p = 0.014) and disease-free survival (log-rank p = 0.0015). However, there was no relationship between sarcopenia and IHC score of Caveolin-1 expression in CAF. Conclusion: The present result suggested that sarcopenia and Caveolin-1 expression of CAF in patient with pancreatic cancer is respectively associated with poor prognosis, but not associated each other. Now, we focus on Caveolin-1 expression of CAF, and establish primary cultures of CAF from the pancreatic cancer tissues and examine its function. Citation Format: Kensuke Yamamura, Yo-ichi Yamashita, Yuki Kitano, Kota Arima, Takayoshi Kaida, Tatsunori Miyata, Shigeki Nakagawa, Kosuke Mima, Katsunori Imai, Daisuke Hashimoto, Akira Chikamoto, Hideo Baba. The association between sarcopenia and cellular senescence of cancer associated fibroblast in pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1770. doi:10.1158/1538-7445.AM2017-1770

Caveolin-1 is Markedly Downregulated in Patients with Early-Stage Colorectal Cancer.

Caveolin-1 (CAV-1), the main scaffold protein in caveolae, is frequently deregulated in human cancer. Of importance, this protein has been described to show tumor suppressor or oncogenic properties depending on the cell type and the stage of the disease. In fact, its role in colorectal cancer (CRC) remains to be fully clarified due to discrepancies in the literature. We analyzed CAV-1 by western blot in a set of early-stage CRC patients with paired tumor tissue and normal colonic mucosa available. CAV-1 mRNA and expression levels of miR-124, 133 and 802 were quantified by real-time PCR. We found CAV-1 strongly downregulated in 76.2% of tumor samples and associated with the subgroup of elderly patients (p=0.027). We observed by real-time PCR a lack of correlation between CAV-1 mRNA and protein levels in some cases with CAV-1 downregulated by western blot, and miR-124 deregulation was identified as a potential contributing alteration to decrease CAV-1 protein expression. CAV-1 is commonly downregulated in patients with primary CRC, which suggests its tumor suppressor role in early stages of this disease. Moreover, based on our findings, the previous discrepancies observed in different studies to date could be due to a complex posttranscriptional CAV-1 regulation.

Role of caveolin-1/vascular endothelial growth factor pathway in basic fibroblast growth factor-induced angiogenesis and neurogenesis after treadmill training following focal cerebral ischemia in rats

Exercise is known to aid functional recovery following ischemia, though the mechanisms responsible for the beneficial effects of exercise on recovery from ischemic stroke are not fully understood. Basic fibroblast growth factor (bFGF) contributes to angiogenesis and promotes neurologic functional recovery after stroke. The present study aimed to investigate the possible mechanisms whereby treadmill exercise ameliorated impaired angiogenesis and neurogenesis following transient cerebral ischemia in middle cerebral artery occlusion (MCAO) rats. Treadmill exercise was started 2days after ischemia-reperfusion in MCAO rats and continued until 7 or 28days after MCAO, after which the animals were sacrificed. Changes in neurological deficit, infarction volume, neuronal morphology, expression levels of bFGF, caveolin-1, and vascular endothelial growth factor (VEGF), and angiogenesis and neurogenesis in the ischemic penumbra were examined by reverse transcription-polymerase chain reaction, western blots, and/or double immunofluorescence. The results suggested that treadmill exercise promoted the expression of bFGF, improved neurological recovery, and reduced infarct volume compared with non-exercised rats, and also enhanced the expression of caveolin-1, VEGF, VEGF receptor 2(FIK-1)/CD34, and Brdu/nestin staining. Small interfering RNA targeting bFGF blocked the protective effects of bFGF. In addition, 4weeks of post-stroke recovery still ameliorated ischemia-induced damage without bFGF shRNA. These findings suggest a novel mechanism underlying the beneficial effects of bFGF following stroke, and indicate that treadmill exercise may aid stroke recovery by regulating the caveolin-1/VEGF pathway in the ischemic zone.

MiR-124-3p attenuates hyperphosphorylation of Tau protein-induced apoptosis via caveolin-1-PI3K/Akt/GSK3β pathway in N2a/APP695swe cells.

Hyperphosphorylation of Tau forming neurofibrillary tangles has been considered as a crucial event in the pathogenesis of Alzheimer's disease (AD). MiR-124-3p belongs to microRNA (miRNA) family and was markedly decreased in AD, however, the functions of miR-124-3p in the pathogenesis of AD remain unknown. We observed that the expression of miR-124-3p was significantly decreased in N2a/APP695swe cells; and transfection of miR-124-3p mimics not only attenuated cell apoptosis and abnormal hyperphosphorylation of Tau protein without any changes of total Tau protein, but also increased expression levels of Caveolin-1, phosphoinositide 3-kinase (PI3K), phospho-Akt (Akt-Ser473)/Akt, phospho-glycogen synthase kinase-3 beta (GSK-3β-Ser9)/GSK-3β in N2a/APP695swe cells. We further found that miR-12-3p directly targeted Caveolin-1; miR-124-3p inhibited abnormal hyperphosphorylation of Tau by regulating Caveolin-1-PI3K/Akt/GSK3β pathway in AD. This study reveals that miR-124-3p may play a neuroprotective role in AD, which may provide new ideas and therapeutic targets for AD.

Expression and clinicopathologic significance of caveolin-1 and erythropoietin producing human hepatocellular receptor A2 in osteosarcoma

Objective To investigate the expression of caveolin-1 and erythropoietin producing human hepatocellular receptor A2 (EphA2) in osteosarcoma, and their relation to the clinicopathological features. Methods Immunohistochemical method was used to assess the expression of caveolin-1 and EphA2 in 23 cases of osteochondroma tissues and 73 cases of osteosarcoma tissues. Results The expression rate of caveolin-1 in osteosarcoma tissues was significantly higher than that in osteochondroma tissues (63.01% vs. 13.04%, P<0.01). The expression levels of caveolin-1 were related to Eneeking stage and lung metastasis of osteosarcoma (P<0.05). The expression rate of EphA2 in osteosarcoma tissues was significantly higher than that in osteochondroma tissues (75.34% vs. 8.70%, P<0.01). The expression levels of EphA2 were related to soft tissue involvement, Eneeking stage and lung metastasis of osteosarcoma (P<0.05). Conclusion Detection of caveolin-1 and EphA2 expression in osteosarcoma may help to predict the biological behaviors of osteosarcoma. Key words: Caveolin-1; Erythropoietin producing human hepatocellular receptor A2; Osteosarcoma