Caveolin-1 Levels Research Articles

Inhibitory effect of paeonol on aortic endothelial inflammation in atherosclerotic rats by up-regulation of caveolin-1 expression and suppression of NF-κB pathway

To explore whether paeonol can play an anti-atherosclerotic role by regulating the expression of aortic caveolin-1 and affecting NF-κB pathway, so as to inhibit the inflammatory response of vascular endothelium in atherosclerotic rats. The atherosclerotic model of rats was induced by high-fat diet and vitamin D_2. The primary culture of vascular endothelial cells(VECs) was carried out by tissue block pre-digestion and adherent method. The injury model of VECs was induced by lipopolysaccharide(LPS), and filipin, a small concave protein inhibitor, was added for control. HE staining was used to observe pathological changes of aorta. TNF-α, IL-6 and VCAM-1 were detected by ELISA. Western blot assay was used to detect the protein expression levels of caveolin-1 and p65 in aorta and VECs. The results showed that as compared with model group, paeonol significantly reduced aortic plaque area and lesion degree in rats, decreased the level of serum TNF-α, IL-6 and VCAM-1 in the rats and enhanced the relative expression level of caveolin-1, decreased p65 expression conversely(P<0.05 or P<0.01). In vitro, as compared to model group, paeonol obviously improved cell morphology, decreased the secretion of TNF-α, IL-6 and VCAM-1 in VECs, increased caveolin-1 expression, and decreased p65 protein expression(P<0.05 or P<0.01). Furthermore, filipin could reverse the effect of paeonol on expression of inflammatory factors and proteins(P<0.05 or P<0.01). According to the results, it was found that paeonol could play the role of anti-atherosclerosis by up-regulating the expression of caveolin-1 and inhibiting the activation of NF-κB pathway to reduce vascular inflammation in atherosclerotic rats.

Clinicopathological and prognostic significance of caveolin-1 and ATG4C expression in the epithelial ovarian cancer.

Altered expression of caveolin-1 (CAV1) and autophagy marker ATG4C is observed in various types of human cancers. However, the clinical significance of CAV1 and ATG4C expression in epithelial ovarian cancer (EOC) remains largely unknown. The present study aims to explore the clinicopathological value and prognostic significance of CAV1 and ATG4C expression in EOC. The expression pattern and prognostic value of CAV1 and ATG4C mRNA in EOC were analyzed using data from the Cancer Genome Atlas (TCGA) database (N = 373). In addition, immunohistochemistry analysis was performed to detect and assay the expression of CAV1 and ATG4C proteins in tissue microarray of EOC. Based on TCGA data, Kaplan-Meier analysis indicated that patients with low CAV1 mRNA (p = 0.021) and high ATG4C mRNA (p = 0.018) expression had a significantly shorter overall survival (OS). Cox regression analysis demonstrated that the expression levels of CAV1 (p = 0.023) and ATG4C mRNA (p = 0.040) were independent prognostic factors for OS in EOC. In addition, the Concordance Index of the nomogram for OS prediction was 0.660. Immunohistochemical analysis showed the expression levels of stromal CAV1 and cancerous ATG4C proteins, and high expression of both CAV1 and ATG4C protein in the stroma were found to significantly correlate with the histologic subtypes of EOC, especially with serous subtype. Decreased expression of CAV1 mRNA and increased expression of ATG4C mRNA in EOC can predict poor overall survival. The expression levels of CAV1 protein in stromal cells and ATG4C protein in cancer cells are significantly associated with histologic subtypes of EOC. These findings suggest that CAV1 and ATG4C serve as useful prognostic biomarkers and candidate therapeutic targets in EOC.

Effect of Aerobic and Resistance Training on Endothelial Progenitor Cells in Mice with Type 2 Diabetes.

Since no study has explored whether exercise could improve impaired proliferation, migration, and angiogenesis of endothelial progenitor cells (EPCs) in animal models or humans with type 2 diabetes, we aimed to explore the effect of different models of exercise on EPC function and expression of caveolin-1, PI3K, and AKT in mice with type 2 diabetes. Male db/db mice (age: 8 weeks) with type 2 diabetes were subjected to aerobic training (AT), resistance training (RT), or combined aerobic and resistance training (AT+RT) 3 or 4 days/week. Mice in the control group remained sedentary with no specific training requirement. Bone marrow-derived EPCs were isolated, and the protein concentrations of caveolin-1, Pi3k, and AKT, and EPC function, were identified in the 1st, 4th, 8th, and 12th weeks of the intervention. Greater increases in proliferation, migration, and angiogenesis were observed in the AT, RT, and AT+RT groups than in the control group. AT+RT was more effective than AT or RT in improving the migratory and angiogenesis function of EPCs in mice with type 2 diabetes and achieved maximum improvement after 8 weeks of intervention. Western blot analysis showed that caveolin-1, p-PI3k, and p-Akt levels were obviously increased in the AT, RT, and AT+RT groups compared with the control group. The expression level of these proteins in the AT+RT group was higher than that in the AT and RT groups. AT+RT may be a helpful reference when choosing exercise methods for the prevention of diabetes-related cardiovascular diseases.

Benzo[a]pyrene exposure induced reproductive endocrine-disrupting effects via the steroidogenic pathway and estrogen signaling pathway in female scallop Chlamys farreri

Benzo[a]pyrene (B[a]P), as one of the typical polycyclic aromatic hydrocarbons and environmental contaminants, may cause endocrine disrupting effects and reproductive impairments in bivalves. However, the molecular mechanisms are still not fully understood. In this study, three reproductive stages (proliferative stage, growing stage and mature stage) of female scallops Chlamys farreri were exposed to B[a]P at 0, 0.38 and 3.8 μg/L. The present study determined the adverse effects of B[a]P on gonadosomatic index, circulating hormone concentrations, endocrine-associated gene expression and ovarian histology. Significant decrease in sex hormones including progesterone (P), testosterone (T) and 17β-estradiol (E2), was observed in B[a]P-treated C. farreri at growing stage and mature stage. These effects were associated with down-regulated expression of steroidogenic enzymes, including 3β-HSD, CYP17 and 17β-HSD, which were regulated by the upstream adenylate cyclase (Adcy) - protein kinase (PKA) signaling pathway. Ovarian transcript levels of estrogen receptor (ER) and caveolin-1 (cav-1) were decreased in B[a]P-treated C. farreri. Vitellogenin (Vtg), an estrogen-mediated gene involved in ovarian development, was down-regulated by B[a]P. Furthermore, ovarian histology was investigated to clarify the impairment of B[a]P on ovaries at growing stage and mature stage. Overall, the present results elucidated the anti-estrogenic mechanisms along the steroidogenic pathway and estrogen signaling pathway for the stage-dependent endocrine-disrupting effects of B[a]P. This finding provides important information regarding to the underlying molecular mechanisms of B[a]P-induced endocrine disruption in different reproductive stages of bivalves. In addition, the adverse effects should be taken into concertation during protection of bivalves germplasm resources and comprehensive evaluation of ecological risks.

Expression of Caveolin-1 Is Associated With Thyroid Function in Patients With Human Papillary Thyroid Carcinoma.

Objective:The aim of this study was to evaluate the levels of caveolin-1 in thyroid follicular epithelial cells of papillary thyroid cancer, follicular thyroid cancer, and nonmalignant thyroid nodule benign follicular adenoma, as well as to explore the relationship between the levels of caveolin-1 and thyroid function.Methods:Thirty cases of papillary thyroid cancer, 10 cases of follicular thyroid cancer, 32 cases of nonmalignant thyroid nodule benign follicular adenoma, and 30 controls were enrolled in this study. Caveolin-1 expression in tissue specimens obtained from these cases was evaluated by immunohistochemistry and Western blotting.Results:Caveolin-1 expression in thyroid epithelial cells of patients with papillary thyroid cancer, particularly female patients, was significantly higher than that in patients with follicular thyroid cancer and nonmalignant thyroid nodule benign follicular adenoma (P < .005). Serum thyroid-stimulating hormone (TSH) levels in the caveolin-1-positive expression group were lower than that in the caveolin-1-negative expression group, and the lowest expression of caveolin-1 was detected in tissues of patients with Graves’ disease. The serum TSH level was associated with caveolin-1 expression in thyroid epithelial cells.Conclusion:Caveolin-1 may participate in regulating thyroid function and is a potential biomarker of follicular thyroid cancer.

Caveolin-1 regulates the ASMase/ceramide-mediated radiation response of endothelial cells in the context of tumor\u2013stroma interactions

The integral membrane protein caveolin-1 (CAV1) plays a central role in radioresistance-mediating tumor–stroma interactions of advanced prostate cancer (PCa). Among the tumor–stroma, endothelial cells (EC) evolved as critical determinants of the radiation response. CAV1 deficiency in angiogenic EC was already shown to account for increased apoptosis rates of irradiated EC. This study explores the potential impact of differential CAV1 levels in EC on the acid sphingomyelinase (ASMase)/ceramide pathway as a key player in the regulation of EC apoptosis upon irradiation and cancer cell radioresistance. Enhanced apoptosis sensitivity of CAV1-deficient EC was associated with increased ASMase activity, ceramide generation, formation of large lipid platforms, and finally an altered p38 mitogen-activated protein kinase (MAPK)/heat-shock protein 27 (HSP27)/AKT (protein kinase B, PKB) signaling. CAV1-deficient EC increased the growth delay of LNCaP and PC3 PCa cells upon radiation treatment in direct 3D spheroid co-cultures. Exogenous C6 and C16 ceramide treatment in parallel increased the growth delay of PCa spheroids and induced PCa cell apoptosis. Analysis of the respective ceramide species in PCa cells with increased CAV1 levels like those typically found in radio-resistant advanced prostate tumors further revealed an upregulation of unsaturated C24:1 ceramide that might scavenge the effects of EC-derived apoptosis-inducing C16 ceramide. Higher ASMase as well as ceramide levels could be confirmed by immunohistochemistry in human advanced prostate cancer specimen bearing characteristic CAV1 tumor–stroma alterations. Conclusively, CAV1 critically regulates the generation of ceramide-dependent (re-)organization of the plasma membrane that in turn affects the radiation response of EC and adjacent PCa cells. Understanding the CAV1-dependent crosstalk between tumor cells and the host-derived tumor microvasculature and its impact on radiosensitivity may allow to define a rational strategy for overcoming tumor radiation resistance improving clinical outcomes by targeting CAV1.

Src-family kinase inhibitors block early steps of caveolin-1-enhanced lung metastasis by melanoma cells

In advanced stages of cancer disease, caveolin-1 (CAV1) expression increases and correlates with increased migratory and invasive capacity of the respective tumor cells. Previous findings from our laboratory revealed that specific ECM-integrin interactions and tyrosine-14 phosphorylation of CAV1 are required for CAV1-enhanced melanoma cell migration, invasion and metastasis in vivo. In this context, CAV1 phosphorylation on tyrosine-14 mediated by non-receptor Src-family tyrosine kinases seems to be important; however, the effect of Src-family kinase inhibitors on CAV1-enhanced metastasis in vivo has not been studied. Here, we evaluated the effect of CAV1 and c-Abl overexpression, as well as the use of the Src-family kinase inhibitors, PP2 and dasatinib (more specific for Src/Abl) in lung metastasis of B16F10 melanoma cells. Overexpression of CAV1 and c-Abl enhanced CAV1 phosphorylation and the metastatic potential of the B16F10 murine melanoma cells. Alternatively, treatment with PP2 or dasatinib for 2 h reduced CAV1 tyrosine-14 phosphorylation and levels recovered fully within 12 h of removing the inhibitors. Nonetheless, pre-treatment of cells with these inhibitors for 2 h sufficed to prevent migration, invasion and trans-endothelial migration in vitro. Importantly, the transient decrease in CAV1 phosphorylation by these kinase inhibitors prevented early steps of CAV1-enhanced lung metastasis by B16F10 melanoma cells injected into the tail vein of mice. In conclusion, this study underscores the relevance of CAV1 tyrosine-14 phosphorylation by Src-family kinases during the first steps of the metastatic sequence promoted by CAV1. These findings open up potential options for treatment of metastatic tumors in patients in which Src-family kinase activation and CAV1 overexpression favor dissemination of cancer cells to secondary sites.

Study of immune responses in mice to oral administration of Flor·Essence.

Flor·Essence (FE), a natural food grade herbal formula product manufactured by Flora Manufacturing & Distributing Ltd., has been used by patients with cancer in North America to stimulate immune cells in order to attenuate or reverse immune damage. To elucidate the mechanisms underlying the effects of FE on the immune system, spleen lymphocyte proliferation was analyzed by an MTT assay, and the phagocytic capacity of macrophages was measured via the neutral red phagocytosis method. The cytotoxicity of natural killer (NK) cells towards K562 cells was assessed via a CytoTox 96 assay. The production of the cytokines interleukin (IL)-12 and interferon (IFN)-γ in the peripheral blood was determined via ELISA and PCR analysis. The expression levels of caveolin-1 and NF-κB were measured via western blotting. In addition, cyclophosphamide was used to establish a mouse model of immunosuppression. It was found that the proliferation of splenocytes, the phagocytic capacity of macrophages and the cytotoxicity of NK cells against K562 cells were increased after oral administration of FE to mice. FE augmented the production of IL-12 and IFN-γ in the peripheral blood of mice. FE significantly increased the expression of proliferating cell nuclear antigen and caveolin-1, and decreased NF-κB expression. Finally, FE enhanced the viability of immune cells from cyclophosphamide-treated immunosuppressed mice. The results indicated that FE could activate immune responses and enhance natural immunity, suggesting that oral administration of FE can activate the body's immune response and resist damage caused by cyclophosphamide chemotherapy.

Caveolin-1 Impacts on TGF-β Regulation of Metabolic Gene Signatures in Hepatocytes.

Caveolin-1 (CAV1) is a membrane protein associated with metabolism in various cell types. The transforming growth factor beta (TGF-β) is a pro-fibrogenic cytokine in the liver, but its metabolic gene signatures remain unclear to date. We have previously shown that CAV1 alters TGF-β signaling and blocks its pro-apoptotic function. Here, we defined TGF-β-induced metabolic gene signatures in hepatocytes and assessed whether CAV1 abundance affects TGF-β control of those metabolic genes. Microarray analyses of primary hepatocytes after TGF-β stimulation (48 h) showed differential expression of 4224 genes, of which 721 are metabolic genes (adjusted p < 0.001). Functional annotation analysis revealed that TGF-β mainly suppresses metabolic gene network, including genes involved in glutathione, cholesterol, fatty acid, and amino acid metabolism. TGF-β also upregulated several genes related to glycan metabolism and ion transport. In contrast to TGF-β effects, CAV1 knockdown triggered the upregulation of metabolic genes. Immortalized mouse hepatocytes (AML12 cells) were used to validate the gene changes induced by TGF-β stimulation and CAV1 knockdown. Noteworthy, of the TGF-β metabolic target genes, CAV1 modulated the expression of 228 (27%). In conclusion, we present several novel metabolic gene signatures of TGF-β in hepatocytes and show that CAV1 abundance alters almost a third of these genes. These findings could enable a better understanding of TGF-β function in normal and diseased liver especially where differential CAV1 level is implicated.

Aerobic and resistance training enhances endothelial progenitor cell function via upregulation of caveolin-1 in mice with type 2 diabetes

BackgroundTo explore the effect of aerobic training (AT), resistance training (RT) or a combination of AT and RT (AT+RT) on the function of endothelial progenitor cells (EPCs) in mice with type 2 diabetes and the potential effective mechanismsMethodsEight-week-old db/db male mice were used as type 2 diabetic animal models in this study. Mice were randomly assigned to the control group (n = 5), AT group (n = 5), RT group (n = 5) and AT+RT group (n = 5). Mice in the control group remained sedentary with no specific training requirement. Mice were motivated to perform AT, RT or AT+RT by a gentle pat on their body for 3 or 4 days/week for 14 days. AT was performed by treadmill running, RT was performed by ladder climbing and AT+RT involved both AT and RT. Bone-derived EPCs were isolated after 14 days of the intervention. EPC expression of CD31, CD34, CD133, CD144 and VEGFR2 was detected by immunofluorescence staining. Fluorescence detection was performed on attached mononuclear cells to detect double-positive EPCs. We then explored the effect of caveolin-1 knockdown (lentiviral vector with caveolin-1-siRNA) on the proliferation and adherence of EPCs and the concentration of caveolin-1 and PI3K/AKT via western blot analyses.ResultsCompared to the mice in the control group, the mice in the AT, RT and AT+RT groups presented significant increases in proliferation and adherence after 14 days of intervention. AT+RT induced an increase in EPC adherence, which was greater than that of the control, RT and AT groups. Caveolin-1 knockdown inhibited the EPC proliferative and adherent abilities. The AT+RT group showed higher levels of caveolin-1 and p-AKT than the control group, but these changes were decreased by caveolin-1-siRNA transfection.ConclusionCombined AT and RT is an effective way to improve EPC function through upregulation of caveolin-1 in mice with type 2 diabetes.

Effect of Minoxidil on Trabecular Outflow via the Paracellular Pathway

PurposeTo investigate the pathway and effects of minoxidil on trabecular outflow in cultured human trabecular meshwork (TM) cells.MethodsAfter exposing primarily cultured TM cells to 0, 10, 50, or 100 µM minoxidil sulfate (MS), trabecular outflow was assessed by measuring TM cell monolayer permeability to carboxyfluorescein and transepithelial electrical resistance. To assess the pathway of permeability changes, caveolin-1, occludin, and claudin-5 levels were measured via western blot. Generation of reactive oxygen species (ROS) was measured using the dichlorofluorescein diacetate assay. To assess the involvement of nitric oxide (NO) in minoxidil-induced permeability increase, the degrees of endothelial nitric oxide synthase mRNA expression and NO production were measured with reverse transcription polymerase chain reaction and Griess assays, respectively. Permeability was also measured with co-exposure to 50 µM N-acetyl cysteine.ResultsMS significantly increased TM cell monolayer permeability (p < 0.05) and decreased transepithelial electrical resistance (p < 0.05). MS decreased the degree of endothelial nitric oxide synthase mRNA expression but did not affect NO production. MS decreased occludin and claudin-5 levels but did not affect caveolin-1 level. MS at 100 µM increased the generation of ROS, and MS-induced permeability increase was attenuated after co-exposure to 50 µM N-acetyl cysteine.ConclusionsMinoxidil may preferentially increase trabecular permeability via a paracellular pathway by downregulation of tight junction proteins. This minoxidil-induced permeability through the TM may be mediated by generation of ROS.

Internalization and Transportation of Endothelial Cell Surface KCa2.3 and KCa3.1 in Normal Pregnancy and Preeclampsia.

Altered redox state modulates the expression levels of endothelial KCa2.3 and KCa3.1 (KCas) in normal pregnancy (NP) and preeclampsia (PE), thereby regulating vascular contractility. The mechanisms underlying KCas endocytosis and transportation remain unknown. We investigated the regulation of KCas expression in plasma membrane (PM) during NP and PE. Cultured human uterine artery endothelial cells were incubated in serum from normal nonpregnant women and women with NP or PE, or in oxidized LDL-, or lysophosphatidylcholine- (LPC-) containing a medium for 24 hours. NP serum elevated PM levels of KCas and reduced caveolin-1 and clathrin levels. PE serum, oxidized LDL, or LPC reduced PM levels of KCas and elevated caveolin-1, clathrin, Rab5c, and early endosome antigen-1 (EEA1) levels. Reduced KCas levels by PE serum or LPC were reversed by inhibition of caveolin-1, clathrin, or EEA1. Catalase and glutathione peroxidase 1 (GPX1) knockdown elevated PM-localized KCas levels and reduced caveolin-1 and clathrin levels. Elevated KCa2.3 levels upon catalase and GPX1 knockdown were reversed by PEG-catalase treatment. An H2O2 donor reduced clathrin and Rab5c. In contrast, elevated clathrin, caveolin-1, or colocalization of caveolin-1 with KCa3.1 by PE serum or LPC was reversed by NADPH oxidase inhibitors or antioxidants. A superoxide donor xanthine+xanthine oxidase elevated caveolin-1 or Rab5c levels. We concluded that KCas are endocytosed in a caveola- or a clathrin-dependent manner and transported in a Rab5c- and EEA1-dependent manner during pregnancy. The endocytosis and transportation processes may slow down via H2O2-mediated pathways in NP and may be accelerated via superoxide-mediated pathways in PE.

Plasma membrane proteome of adhesion-competent endometrial epithelial cells and its modulation by Rab11a.

Human endometrial epithelium (EE) is composed of a multitude of proteins, amongst which those localized on the plasma membrane [plasma membrane proteins (PMPs)] are of critical relevance in the early stages of implantation. Evidence supports the key role offew PMPs in implantation. However, many remain unidentified, as efforts have not been made till date to generate the plasma membrane proteome of human EE cells, using a gel-free approach. This study presents a protein catalog of the PMP enriched fraction of Ishikawa cell line; often used as an in vitro model for embryo-adhesive EE. Liquid chromatography with tandem mass spectrometry identified 3,598 proteins. Of these, 1,963 proteins were annotated for their membrane localization. Of 1,963 proteins, 1,321 were found to have a transmembrane domain and 43 proteins had glycophosphatidylinositol (GPI) anchor. Extensive data mining revealed endometrial expression of 943 proteins reported in humans and/or rodents. Further, quantitative alterations were observed in the plasma membrane proteome on the perturbation of intracellular trafficking. Silencing of Rab11a (known for its role in plasma membrane organization) expression caused alteration in the abundance of 74 proteins. Caveolin-1 and EpCAM levels were reduced whereas Rab4a abundance increased in the PMP extracts of Rab11a deficient cells, compared with control cells. Briefly, the study reports the identity of several novel plasma membrane-localized proteins. A major spin-off of the study is the identification of novel proteins trafficked by Rab11a to the plasma membrane. Targeted analysis of novel PMPs may reveal their specific roles in endometrial receptivity and implantation.

Caveolin-1 Expression Together with VEGF can be a Predictor for Lung Metastasis and Poor Prognosis in Osteosarcoma.

Caveolin-1, the major protein component of caveolae, plays vital functions in tumorigenesis and metastasis. Previous evidence demonstrated the positive role of Caveolin-1 in the regulation of endothelial cell differentiation and the involvement of Caveolin-1 in vascular endothelial growth factor (VEGF) mediated angiogenesis. The correlation of Caveolin-1 expression and angiogenesis is not yet elucidated in osteosarcoma. This study aimed to investigate the expression levels of Caveolin-1 and VEGF in osteosarcoma and their associations with clinicopathological data. This study included 66 formalin-fixed and paraffin embedded osteosarcoma tissue samples. The expression levels of Caveolin-1 and VEGF were assessed by immunohistochemistry. Then associations with clinicopathological variables and the correlation between both markers were evaluated statistically. We also investigated the expression of Caveolin-1 and VEGF values in gene microarrays of osteosarcoma patients and cell lines by using GEO data sets on https://www.ncbi.nlm.nih.gov. Caveolin-1 and VEGF were expressed in 19.6% and 77.3%, respectively. Caveolin-1 expression was associated positively with osteoblastic histological subtype (P < 0.0001). VEGF expression showed positive association with patient age, histological grade and clinical stage (P = 0.031, P = 0.024 and P < 0.001; respectively). An inverse correlation between Caveolin-1 and VEGF expressions in osteosarcoma was found (r = 0.2 P = 0.04). In silico analysis of Caveolin-1 and VEGF expression supported our results. Our results suggest that Caveolin-1 may act as a tumor suppressor in osteosarcoma. Down-regulation of Caveolin-1 can be used as an indicator for poor prognosis in osteosarcoma patients. Meanwhile, overexpression of VEGF is a predictor of pulmonary metastasis and poor prognosis.

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.

Depletion of Caveolin-1 in Type 2 Diabetes Model Induces Alzheimer's Disease Pathology Precursors.

Type 2 diabetes mellitus (T2DM) is a risk factor for the development of late-onset Alzheimer's disease (AD). However, the mechanism underlying the development of late-onset AD is largely unknown. Here we show that levels of the endothelial-enriched protein caveolin-1 (Cav-1) are reduced in the brains of T2DM patients compared with healthy aging, and inversely correlated with levels of β-amyloid (Aβ). Depletion of Cav-1 is recapitulated in the brains of db/db (Leprdb ) diabetic mice and corresponds with recognition memory deficits as well as the upregulation of amyloid precursor protein (APP), BACE-1, a trending increase in β-amyloid Aβ42/40 ratio and hyperphosphorylated tau (p-tau) species. Importantly, we show that restoration of Cav-1 levels in the brains of male db/db mice using adenovirus overexpressing Cav-1 (AAV-Cav-1) rescues learning and memory deficits and reduces pathology (i.e., APP, BACE-1 and p-tau levels). Knocking down Cav-1 using shRNA in HEK cells expressing the familial AD-linked APPswe mutant variant upregulates APP, APP carboxyl terminal fragments, and Aβ levels. In turn, rescue of Cav-1 levels restores APP metabolism. Together, these results suggest that Cav-1 regulates APP metabolism, and that depletion of Cav-1 in T2DM promotes the amyloidogenic processing of APP and hyperphosphorylation of tau. This may suggest that depletion of Cav-1 in T2DM underlies, at least in part, the development of AD and imply that restoration of Cav-1 may be a therapeutic target for diabetic-associated sporadic AD.SIGNIFICANCE STATEMENT More than 95% of the Alzheimer's patients have the sporadic late-onset form (LOAD). The cause for late-onset Alzheimer's disease is unknown. Patients with Type 2 diabetes mellitus have considerably higher incidence of cognitive decline and AD compared with the general population, suggesting a common mechanism. Here we show that the expression of caveolin-1 (Cav-1) is reduced in the brain in Type 2 diabetes mellitus. In turn, reduced Cav-1 levels induce AD-associated neuropathology and learning and memory deficits. Restoration of Cav-1 levels rescues these deficits. This study unravels signals underlying LOAD and suggests that restoration of Cav-1 may be an effective therapeutic target.

MiR-124 Regulates Caveolin-1 Expression and Affects Proliferation and Apoptosis of Osteosarcoma Cells

Osteosarcoma (OS) seriously affects human health. miR-124 expression is closely related to osteosarcoma, but its specific mechanism remains unclear. Our study intends to evaluate miR-124’s effect on osteosarcoma. MG-63 cells were transfected with miR-124 mimics/NC followed by analysis of miR-124 expression by real-time PCR, cell proliferation by CCK8 assay, cell apoptosis by flow cytometry as well as the level of caveolin-1 (CAV1) by Western blot. miR-124 was significantly lower and CAV1 was increased in the four osteosarcoma cells than those in normal osteoblasts (P &lt; 0.05). miR-124 mimics transfection significantly reduced CAV1 level and cell number (P &lt; 0.05) and increased cell apoptosis rate (P &lt; 0.05). Moreover, miR-124 inhibitor significantly promoted the relative luciferase activity in pmirGLO-CAV1-3′UTR-wt-transfected cells (P &lt; 0.05). miR-124 affects osteosarcoma cell proliferation and apoptosis via targeting CAV1.

Immunoreactivity for prostate specific antigen and Ki67 differentiates subgroups of prostate cancer related to outcome

Based on gene-expression profiles, prostate tumors can be subdivided into subtypes with different aggressiveness and response to treatment. We investigated if similar clinically relevant subgroups can be identified simply by the combination of two immunohistochemistry markers: one for tumor cell differentiation (prostate specific antigen, PSA) and one for proliferation (Ki67). This was analyzed in men with prostate cancer diagnosed at transurethral resection of the prostate 1975–1991 (n = 331) where the majority was managed by watchful waiting. Ki67 and PSA immunoreactivity was related to outcome and to tumor characteristics previously associated with prognosis. Increased Ki67 and decreased PSA were associated with poor outcome, and they provided independent prognostic information from Gleason score. A combinatory score for PSA and Ki67 immunoreactivity was produced using the median PSA and Ki67 levels as cut-off (for Ki67 the upper quartile was also evaluated) for differentiation into subgroups. Patients with PSA low/Ki67 high tumors showed higher Gleason score, more advanced tumor stage, and higher risk of prostate cancer death compared to other patients. Their tumor epithelial cells were often ERG positive and expressed higher levels of ErbB2, phosphorylated epidermal growth factor receptor (pEGF-R) and protein kinase B (pAkt), and their tumor stroma showed a reactive response with type 2 macrophage infiltration, high density of blood vessels and hyaluronic acid, and with reduced levels of caveolin-1, androgen receptors, and mast cells. In contrast, men with PSA high/Ki67 low tumors were characterized by low Gleason score, and the most favorable outcome amongst PSA/Ki67-defined subgroups. Men with PSA low/Ki67 low tumors showed clinical and tumor characteristics intermediate of the two groups above. A combinatory PSA/Ki67 immunoreactivity score identifies subgroups of prostate cancers with different epithelial and stroma phenotypes and highly different outcome but the clinical usefulness of this approach needs to be validated in other cohorts.

The CAV1 Gene 3' Untranslated Region Single Nucleotide Polymorphisms Are Associated with the Risk of Pulmonary Hypertension in Chinese Han Chronic Obstructive Pulmonary Patients.

Aims: Caveolin-1, which is encoded by the caveolin-1 (CAV1) gene, plays an important role in the development of pulmonary hypertension (PH). The purpose of this study was to determine the relationship between 3' untranslated region (UTR) single nucleotide polymorphisms (SNPs) of the CAV1 gene and the risk of PH in Chinese Han patients with chronic obstructive pulmonary disease (COPD). Methods: From March 2016 to October 2018, 235 patients with COPD combined with PH (COPD/PH+) and 240 patients with COPD and without PH (COPD/PH-) were recruited to the study. The CAV1 gene rs1049314, rs8713, rs1049334, rs6867, and rs1049337 loci were genotyped and the plasma hsa-miR-451 and caveolin-1 levels were measured in all subjects. Results: The risk of PH in patients with COPD carrying the C allele of the rs8713 locus was 2.82 times higher than that in A allele carriers (95% confidence interval [CI], 1.94-4.08; p < 0.001). The risk of PH in patients with COPD carrying the T allele of the rs1049337 locus was significantly lower than that in C allele carriers (odds ratio [OR], 0.48; 95% CI, 0.37-0.63; p < 0.001). The ACGAC haplotype was found to be a highly-significant risk factor for COPD combined with PH (OR, 2.24; 95% CI, 1.20-4.17; p = 0.01). Plasma levels of hsa-miR-451 and the caveolin1 protein in patients with the rs8713 C allele were significantly lower than in those with the wild type (WT) allele regardless of PH status. Conversely, the hsa-miRN-451 and caveolin-1 levels in patients with the rs1049337 mutant C allele were significantly higher than those in the WT T allele (p < 0.05). There was a positive correlation between plasma hsa-miR-451 and caveolin-1 levels in patients with COPD/PH+ and COPD/PH- (r = 0.72 and 0.63, respectively). Conclusion: SNPs of the CAV1 gene loci rs8713 and rs1049337 are associated with a risk of PH in COPD patients. The underlying mechanism is likely to be related to the effect of the SNPs on the regulation of caveolin-1 by hsa-miR-451.

Dasatinib, paclitaxel and carboplatin in women with advanced-stage and recurrent endometrial cancer: A pilot clinical and translational study

Abstract Purpose To evaluate the effect of dasatinib therapy on EphA2 signaling in cancers of women with measurable (biopsy amenable) advanced-stage, chemo-naive primary or recurrent endometrial cancer. Preliminary efficacy was also assessed. Patients and methods We performed a pilot study of single-agent dasatinib lead-in, followed by triplet dasatinib, paclitaxel, and carboplatin. We measured the downstream effectors of EphA2 signaling in pre- and post-dasatinib treatment biopsy tissue samples; we also determined the severity of adverse events and patients' progression-free survival and overall survival durations. Results Eighteen patients were recruited and given dasatinib (150 mg orally daily for 14 days), followed by paclitaxel, carboplatin and dasatinib (daily) for six cycles (21-day cycles). Seventeen patients were evaluable for toxicity and 11 patients for response. A reverse phase protein array and proximity ligation assay revealed that CRAF/BRAF dimerization, caveolin-1 level, and Notch pathway signaling were predictive of response and resistance to dasatinib. Overall, the objective response rate was 45% (95% CI: 17%–77%), with median progression-free survival duration of 10.5 months and median overall survival duration of 30.4 months. The most common grade 3 or 4 adverse events were neutropenia (76%), thrombocytopenia (53%), anemia (53%), and fatigue (12%). Conclusions Caveolin-1 expression, in combination with CRAF/BRAF heterodimerization, is associated with resistance to EphA2 targeting by dasatinib. The triplet combination showed interesting clinical activity in endometrial cancer with acceptable toxicity. Pretreatment with dasatinib may accentuate combination therapy toxicity.