作者机构：
[Tang, Chao-Ke; Yu, Xiao-Hua] Univ South China, Life Sci Res Ctr, Hengyang 421001, Hunan, Peoples R China.;[Tang, Chao-Ke; Yin, Kai] Univ South China, Key Lab Atherosclerol Hunan Prov, Inst Cardiovasc Res, Hengyang 421001, Hunan, Peoples R China.;[Fu, Yu-Chang] Univ Alabama Birmingham, Dept Nutr Sci, Birmingham, AL 35294 USA.;[Zhang, Da-Wei] Univ Alberta, Dept Pediat, Edmonton, AB T6G 2S2, Canada.;[Zhang, Da-Wei] Univ Alberta, Grp Mol & Cell Biol Lipids, Edmonton, AB T6G 2S2, Canada.
通讯机构：
[Yin, Kai] Univ South China, Key Lab Atherosclerol Hunan Prov, Inst Cardiovasc Res, Hengyang 421001, Hunan, Peoples R China.
关键词：
ABCA1;ABCG1;ACAT1;AGE;ATP-binding cassette transporter A1;ATP-binding cassette transporter G1;Atherosclerosis;CD36;CE;ERK1/2;FC;Foam cells;HDL;LDLR;LXR;MAPK;PI3K;PKB;PKC;PPAR response elements;PPAR-gamma;PPREs;RCT;RXR;SR-A;SR-BI;TGF-beta;acyl coenzyme A:cholesterol acyltransferase-1;advanced glycation end products;apoA-I;apoE;apolipoprotein A-I;apolipoprotein E;cholesterol ester;extracellular signal-regulated kinases 1 and 2;free cholesterol;high-density lipoprotein;liver X receptor;low-density lipoprotein receptor;mitogen-activated protein kinase;nCEH;neutral cholesteryl ester hydrolase;ox-LDL;oxidized low-density lipoprotein;peroxisome proliferator-activated receptor-gamma;phosphatidylinositol 3-kinase;protein kinase B;protein kinase C;retinoid X receptor;reverse cholesterol transport;scavenger receptor BI;scavenger receptor class A;transforming growth factor-beta
摘要：
Atherosclerosis is a chronic disease characterized by the deposition of excessive cholesterol in the arterial intima. Macrophage foam cells play a critical role in the occurrence and development of atherosclerosis. The generation of these cells is associated with imbalance of cholesterol influx, esterification and efflux. CD36 and scavenger receptor class A (SR-A) are mainly responsible for uptake of lipoprotein-derived cholesterol by macrophages. Acyl coenzyme A:cholesterol acyltransferase-1 (ACAT1) and neutral cholesteryl ester hydrolase (nCEH) regulate cholesterol esterification. ATP-binding cassette transporters A1 (ABCA1), ABCG1 and scavenger receptor BI (SR-BI) play crucial roles in macrophage cholesterol export When inflow and esterification of cholesterol increase and/or its outflow decrease, the macrophages are ultimately transformed into lipid-laden foam cells, the prototypical cells in the atherosclerotic plague. The aim of this review is to describe what is known about the mechanisms of cholesterol uptake, esterification and release in macrophages. An increased understanding of the process of macrophage foam cell formation will help to develop novel therapeutic interventions for atherosclerosis. (c) 2013 The Authors. Published by Elsevier B.V. All rights reserved.
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience,2012年32(9):3044-3057 ISSN：1529-2401
通讯作者：
Liu, Wenlan
作者机构：
[Liu, Jie; Jin, Xinchun; Liu, Ke J.; Liu, Wenlan] Univ New Mexico, Coll Pharm, Hlth Sci Ctr, Albuquerque, NM 87131 USA.;[Liu, Ke J.] Univ New Mexico, Dept Neurol, Hlth Sci Ctr, Albuquerque, NM 87131 USA.;[Liu, Jie] Univ South China, Sch Med, Dept Med Microbiol & Immunol, Hengyang 421001, Hunan, Peoples R China.
通讯机构：
[Liu, Wenlan] Univ New Mexico, Coll Pharm, Hlth Sci Ctr, Albuquerque, NM 87131 USA.
摘要：
Blood-brain barrier (BBB) disruption occurs early enough to be within the thrombolytic time window, and this early ischemic BBB damage is closely associated with hemorrhagic transformation and thus emerging as a promising target for reducing the hemorrhagic complications of thrombolytic stroke therapy. However, the mechanisms underlying early ischemic BBB damage remain poorly understood. Here, we investigated the early molecular events of ischemic BBB damage using in vitro oxygen-glucose deprivation (OGD) and in vivo rat middle cerebral artery occlusion (MCAO) models. Exposure of bEND3 monolayer to OGD for 2 h significantly increased its permeability to FITC-labeled dextran and promoted the secretion of metalloproteinase-2 and -9 (MMP-2/9) and cytosolic translocation of caveolin-1 (Cav-1). This same OGD treatment also led to rapid degradation of tight junction protein occludin and dissociation of claudin-5 from the cytoskeleton, which contributed to OGD-induced endothelial barrier disruption. Using selective MMP-2/9 inhibitor SB-3CT (2-[[(4-phenoxyphenyl)sulfonyl]methyl]-thiirane) or their neutralizing antibodies or Cav-1 siRNA, we found that MMP-2 was the major enzyme mediating OGD-induced occludin degradation, while Cav-1 was responsible for claudin-5 redistribution. The interaction between Cav-1 and claudin-5 was further confirmed by coimmunoprecipitation. Consistent with these in vitro findings, we observed fluorescence tracer extravasation, increased gelatinolytic activity, and elevated interstitial MMP-2 levels in ischemic subcortical tissue after 2 h MCAO. Moreover, occludin protein loss and claudin-5 redistribution were detected in ischemic cerebromicrovessels. These data indicate that cerebral ischemia initiates two rapid parallel processes, MMP-2-mediated occludin degradation and Cav-1-mediated claudin-5 redistribution, to cause BBB disruption at early stroke stages relevant to acute thrombolysis.
期刊：
Journal of Cellular Biochemistry,2011年112(9):2508-2517 ISSN：1097-4644
通讯作者：
Xiao, Zhi-Qiang
作者机构：
[Wan, Xun-Xun; Qu, Jia-Quan; Zhu, Wei; Yi, Hong; He, Qiu-Yan; Zuo, Jian-Hong; Li, Mao-Yu; Xiao, Zhi-Qiang; Li, Xin-Hui; Zeng, Gu-Qing; Li, Jian-Huang; Chen, Yu] Cent S Univ, Xiangya Hosp, Key Lab Canc Prote, Chinese Minist Hlth, Changsha 410008, Hunan, Peoples R China.;[Zuo, Jian-Hong] Univ S China, Dept Microbiol & Immunol, Sch Med, Hengyang 421001, Peoples R China.;[Zhu, Wei] Univ S China, Affiliated Hosp 1, Dept Oncol, Hengyang 421001, Peoples R China.;[Xiao, Zhi-Qiang] Cent S Univ, Xiangya Hosp, Key Lab Canc Prote, Chinese Minist Hlth, 87 Xiangya Rd, Changsha 410008, Hunan, Peoples R China.
通讯机构：
[Xiao, Zhi-Qiang] Cent S Univ, Xiangya Hosp, Key Lab Canc Prote, Chinese Minist Hlth, 87 Xiangya Rd, Changsha 410008, Hunan, Peoples R China.
关键词：
EGFR;HNSCC;Invasion;MMP‐9;E‐cadherin
摘要：
EGFR is a potent stimulator of invasion and metastasis in head and neck squamous cell carcinomas (HNSCC). However, the mechanism by which EGFR may stimulate tumor cell invasion and metastasis still need to be elucidated. In this study, we showed that activation of EGFR by EGF in HNSCC cell line SCC10A enhanced cell migration and invasion, and induced loss of epitheloid phenotype in parallel with downregulation of E-cadherin and upregulation of N-cadherin and vimentin, indicating that EGFR promoted SCC10A cell migration and invasion possibly by an epithelial to mesenchymal transition (EMT)-like phenotype change. Interestingly, activation of EGFR by EGF induced production of matrix metalloproteinase-9 (MMP-9) and soluble E-cadherin (sE-cad), and knockdown of MMP-9 by siRNA inhibited sE-cad production induced by EGF in SCC10A. Moreover, both MMP-9 knockdown and E-cadherin overexpression inhibited cell migration and invasion induced by EGF in SCC10A. The results indicate that EGFR activation promoted cell migration and invasion through inducing MMP-9-mediated degradation of E-cadherin into sE-cad. Pharmacologic inhibition of EGFR, MEK, and PI3K kinase activity in SCC10A reduced phosphorylated levels of ERK-1/2 and AKT, production of MMP-9 and sE-cad, cell migration and invasion, and expressional changes of EMT markers (E-cadherin and N-cadherin) induced by EGF, indicating that EGFR activation promotes cell migration and invasion via ERK1/2 and PI3K-regulated MMP-9/E-cadherin signaling pathways. Taken together, the data suggest that EGFR activation promotes HNSCC SCC10A cell migration and invasion by inducing EMT-like phenotype change and MMP-9-mediated degradation of E-cadherin into sE-cad related to activation of ERK-1/2 and PI3K signaling pathways. J. Cell. Biochem. 112: 2508-2517, 2011. (C) 2011 Wiley-Liss, Inc.
期刊：
Clinical Cancer Research,2013年19(20):5602-5612 ISSN：1078-0432
通讯作者：
Xie, Xiaoming
作者机构：
[Tang, Hailin; Xie, Xinhua; Xie, Xiaoming; Kong, Yanan; Guo, Jiaoli; Ye, Feng] Sun Yat Sen Univ, Dept Breast Oncol, Ctr Canc, Guangzhou 510060, Guangdong, Peoples R China.;[Tang, Hailin; Xie, Xinhua; Xie, Xiaoming; Kong, Yanan; Guo, Jiaoli; Ye, Feng] Sun Yat Sen Univ, Ctr Canc, State Key Lab Oncol South China, Guangzhou 510060, Guangdong, Peoples R China.;[Deng, Min] Guangzhou Med Univ, Canc Hosp, Guangzhou, Guangdong, Peoples R China.;[Deng, Min] Guangzhou Med Univ, Canc Res Inst, Guangzhou, Guangdong, Peoples R China.;[Tang, Hailin; Tang, Yunyun; Deng, Min; Su, Qi] Univ South China, Canc Res Inst, Hengyang, Hunan, Peoples R China.
通讯机构：
[Xie, Xiaoming] Sun Yat Sen Univ, Dept Breast Oncol, State Key Lab Oncol South China, Ctr Canc, 651 East Dongfeng Rd, Guangzhou 510060, Guangdong, Peoples R China.
通讯机构：
[Xiao, Xianzhong] Cent S Univ, Xiangya Sch Med, Dept Pathophysiol, 110 Xiangya Rd, Changsha 410078, Hunan, Peoples R China.
关键词：
Apoptosis;Doxorubicin;Resveratrol;SIRT1;p53
摘要：
Aims Doxorubicin (DOX) is an anthracycline drug with a wide spectrum of clinical antineoplastic activity, but increased apoptosis has been implicated in its cardiotoxicity. Resveratrol (RES) was shown to harbour major health benefits in diseases associated with oxidative stress. In this study, we aimed to determine the effect of RES on DOX-induced myocardial apoptosis in mice. Methods and results Male Balb/c mice were randomized to one of the following four treatments: saline, RES, DOX, or RES plus DOX (10 mice in each group). DOX treatment markedly depressed cardiac function, decreased the heart weight, the body weight, and the ratio of heart weight to body weight, but inversely increased the level of protein carbonyl, malondialdehyde, and serum lactate dehydrogenase, and induced mitochondrial cytochrome c release and cardiomyocyte apoptosis. However, these effects of DOX were ameliorated by its combination with RES. Further studies with a co-immunoprecipitation assay revealed an interaction between p53 and Sirtuin 1 (SIRT1). It was found by western blot and electrophoretic mobility shift assay that DOX treatment increased p53 protein acetylation and cytochrome c release from mitochondria, activated p53 binding at the Bax promoter, and up-regulated Bax expression, but supplementation with RES could weaken all these effects. Conclusion The protective effect of RES against DOX-induced cardiomyocyte apoptosis is associated with the up-regulation of SIRT1-mediated p53 deacetylation.
摘要：
Recently, microRNAs have been detected in serum and plasma, and circulating microRNA (miRNA) profiles have now been associated with many diseases such as cancers and heart disease, as well as altered physiological states. Because of their stability and disease resistance, circulation miRNAs appear to be an ideal material for biomarkers of diseases and physiological states in blood. However, the lack of a suitable internal reference gene (internal reference miRNA) has hampered research and application of circulating miRNAs. Currently, U6 and miR-16 are the most common endogenous controls in the research of miRNAs in tissues and cells. We performed microarray-based serum miRNA profiling on the serum of 20 nasopharyngeal carcinoma patients and 20 controls to detect the expressions of U6 and miRNAs. Profiling was followed by real-time quantitative Polymerase Chain Reaction (qPCR) in 80 patients (20 each with gastric cancer, nasopharyngeal carcinoma, colorectal cancer, and breast cancer) and 30 non-cancerous controls. qPCR was also performed to detect miRNAs in serum with repeated freezing and thawing. The results of microarray showed that with the exception of U6, Ct values of miR-16, miR-24, miR-142-3p, miR-19b and miR-192 in serum samples of nasopharyngeal carcinoma were greater than control samples. The results of 110 cases showed large fluctuations in U6 expression. The difference between the greatest and the least levels of expression was 3.29 for delta Ct values, and 1.23 for miR-16. The expressions of U6, miR-16 and miR-24 in serum subjected to different freeze-thaw cycles showed that U6 expression gradually decreased after 1, 2, and 4 cycles of freezing and thawing, while the expression of miR-16 and miR24 remained relatively stable. Collectively, our results suggested that U6 is unsuitable as an internal reference gene in the research of circulating miRNAs. (C) 2014 Elsevier Inc. All rights reserved.
摘要：
Background MicroRNAs have been considered as a kind of potential novel biomarker for cancer detection due to their remarkable stability in the blood and the characteristics of their expression profile in many diseases. Methods We performed microarray-based serum miRNA profiling on the serum of twenty nasopharyngeal carcinoma patients at diagnosis along with 20 non-cancerous individuals as controls. This was followed by a real-time quantitative Polymerase Chain Reaction (RT-qPCR) in a separate cohort of thirty patients with nasopharyngeal carcinoma and thirty age- matched non-cancerous volunteers. A model for diagnosis was established by a conversion of mathematical calculation formula which has been validated by analyzing 74 cases of patients with nasopharyngeal carcinoma and 57 cases of non-cancerous volunteers. Results The profiles showed that 39 and 17 miRNAs are exclusively expressed in the serum of non-cancerous volunteers and of patients with nasopharyngeal carcinoma respectively. 4 miRNAs including miR-17, miR-20a, miR-29c, and miR-223 were found to be expressed differentially in the serum of NPC compared with that of non-cancerous control. Based on this, a diagnosis equation with Ct difference method has been established to distinguish NPC cases and non-cancerous controls and validated with high sensitivity and specificity. Conclusions We demonstrate that the serum miRNA-based biomarker model become a novel tool for NPC detection. The circulating 4-miRNA-based method may provide a novel strategy for NPC diagnosis.
摘要：
Rationale: Macrophage cholesterol homeostasis maintenance is the result of a balance between influx, endogenous synthesis, esterification/hydrolysis and efflux. Excessive accumulation of cholesterol leads to foam cell formation, which is the major pathology of atherosclerosis. Previous studies have shown that miR-27 (miR-27a and miR-27b) may play a key role in the progression of atherosclerosis. Objective: We set out to investigate the molecular mechanisms of miR-27a/b in intracellular cholesterol homeostasis. Methods and results: In the present study, our results have shown that the miR-27 family is highly conserved during evolution, present in mammals and directly targets the 3' UTR of ABCA1, LPL, and ACAT1. apoA1, ABCG1 and SR-B1 lacking miR-27 bind sites should not be influenced by miR-27 directly. miR-27a and miR-27b directly regulated the expression of endogenous ABCA1 in different cells. Treatment with miR-27a and miR-27b mimics reduced apoA1-mediated cholesterol efflux by 33.08% and 44.61% in THP-1 cells, respectively. miR-27a/b also regulated HDL-mediated cholesterol efflux in THP-1 macrophages and affected the expression of apoA1 in HepG2 cells. However, miR-27a/b had no effect on total cellular cholesterol accumulation, but regulated the levels of cellular free cholesterol and cholesterol ester. We further found that miR-27a/b regulated the expression of LPL and CD36, and then affected the ability of THP-1 macrophages to uptake Dil-oxLDL. Finally, we identified that miR-27a/b regulated cholesterol ester formation by targeting ACAT1 in THP-1 macrophages. Conclusion: These findings indicate that miR-27a/b affects the efflux, influx, esterification and hydrolysis of cellular cholesterol by regulating the expression of ABCA1, apoA1, LPL, CD36 and ACAT1. (C) 2014 Elsevier Ireland Ltd. All rights reserved.
期刊：
Journal of Experimental & Clinical Cancer Research,2017年36(1):1-12 ISSN：1756-9966
通讯作者：
Zeng, Zhaoyang;Tang, Hailin
作者机构：
[Li, Xiaoling; Zeng, Zhaoyang; He, Rongfang; Li, Guiyuan; Xiong, Wei] Cent S Univ, Xiangya Hosp, Chinese Minist Hlth, Key Lab Carcinogenesis, Changsha, Hunan, Peoples R China.;[Li, Xiaoling; Zeng, Zhaoyang; He, Rongfang; Li, Guiyuan; Xiong, Wei] Cent S Univ, Canc Res Inst, Chinese Minist Educ, Key Lab Carcinogenesis & Canc Invas, Changsha, Hunan, Peoples R China.;[He, Rongfang] Univ South China, Affiliated Hosp 1, Dept Pathol, Hengyang, Hunan, Peoples R China.;[Tang, Hailin; Xie, Xiaoming; Liu, Peng] Sun Yat Sen Univ, Canc Ctr, Dept Breast Oncol, Guangzhou, Guangdong, Peoples R China.;[Tang, Hailin; Xie, Xiaoming; Liu, Peng] State Key Lab Oncol South China, Guangzhou, Guangdong, Peoples R China.
通讯机构：
[Zeng, Zhaoyang] Cent S Univ, Xiangya Hosp, Chinese Minist Hlth, Key Lab Carcinogenesis, Changsha, Hunan, Peoples R China.;[Zeng, Zhaoyang] Cent S Univ, Canc Res Inst, Chinese Minist Educ, Key Lab Carcinogenesis & Canc Invas, Changsha, Hunan, Peoples R China.;[Tang, Hailin] Sun Yat Sen Univ, Canc Ctr, Dept Breast Oncol, Guangzhou, Guangdong, Peoples R China.;[Tang, Hailin] State Key Lab Oncol South China, Guangzhou, Guangdong, Peoples R China.;[Tang, Hailin] Collaborat Innovat Ctr Canc Med, Guangzhou, Guangdong, Peoples R China.
关键词：
Circular RNAs;miR-34a;GFRA1;Competitive endogenous RNAs;Triple negative breast cancer
摘要：
Accumulating evidences indicate that circular RNAs (circRNAs), a class of non-coding RNAs, play important roles in tumorigenesis. However, the function of circRNAs in triple negative breast cancer (TNBC) is largely unknown. We performed circRNA microarrays to identify circRNAs that are aberrantly expressed in TNBC cell lines. Expression levels of a significantly upregulated circRNA, circGFRA1, was detected by quantitative real-time PCR (qRT-PCR) in TNBC cell lines and tissues. Kaplan-Meier survival analysis was used to explore the significance of circGFRA1 in clinical prognosis. Then, we examined the functions of circGFRA1 in TNBC by cell proliferation, apoptosis and mouse xenograft assay. In addition, luciferase assay was used to explore the miRNA sponge function of circGFRA1 in TNBC. Microarray analysis and qRT-PCR verified a circRNA termed circGFRA1 that was upregulated in TNBC. Kaplan-Meier survival analysis showed that upregulated circGFRA1 was correlated with poorer survival. Knockdown of circGFRA1 inhibited proliferation and promoted apoptosis in TNBC. Via luciferase reporter assays, circGFRA1 and GFRA1 was observed to directly bind to miR-34a. Subsequent experiments showed that circGFRA1 and GFRA1 regulated the expression of each other by sponging miR-34a. Taken together, we conclude that circGFRA1 may function as a competing endogenous RNA (ceRNA) to regulate GFRA1 expression through sponging miR-34a to exert regulatory functions in TNBC. circGFRA1 may be a diagnostic biomarker and potential target for TNBC therapy.
作者机构：
[Tang, Chao-ke; Yin, Kai] Univ S China, Inst Cardiovasc Res, Life Sci Res Ctr, Key Lab Atherosclerol Hunan Prov, Hengyang 421001, Hunan, Peoples R China.;[Liao, Duan-fang] Hunan Univ Chinese Med, Sch Pharm, Dept Tradit Chinese Diagnost, Changsha, Hunan, Peoples R China.
通讯机构：
[Tang, Chao-ke] Univ S China, Inst Cardiovasc Res, Life Sci Res Ctr, Key Lab Atherosclerol Hunan Prov, Hengyang 421001, Hunan, Peoples R China.
摘要：
Atherosclerosis is characterized by a chronic inflammatory condition that involves numerous cellular and molecular inflammatory components. A wide array of inflammatory mediators, such as cytokines and proteins produced by macrophages and other cells, play a critical role in the development and progression of the disease ATP-binding membrane cassette transporter A1 (ABCA1) is crucial for cellular cholesterol efflux and reverse cholesterol transport (RCT) and is also identified as an important target in antiatherosclerosis treatment Evidence from several recent studies indicates that inflammation, along with other atherogenic-related mediators, plays distinct regulating roles in ABCA1 expression Proatherogenic cytokines such as interferon (IFN)-gamma and interleukin (IL)-1 beta have been shown to inhibit the expression of ABCA1, while antiatherogenic cytokines, including IL-10 and transforming growth factor (TGF)-beta 1, have been shown to promote the expression of ABCA1 Moreover, some cytokines such as tumor necrosis factor (TNF)-alpha seem to regulate ABCA1 expression in species-specific and dose-dependent manners Inflammatory proteins such as C-reactive protein (CRP) and cyclooxygenase (COX)-2 are likely to inhibit ABCA1 expression during inflammation, and inflammation induced by lipopolysaccharide (LPS) was also found to block the expression of ABCA1 Interestingly, recent experiments revealed ABCA1 can function as an antiinflammatory receptor to suppress the expression of inflammatory factors, suggesting that ABCA1 may be the molecular basis for the interaction between inflammation and ROT. This review aims to summarize recent findings on the role of inflammatory cytokines, inflammatory proteins, inflammatory lipids, and the endotoxin-mediated inflammatory process in expression of ABCA1. Also covered is the current understanding of the function of ABCA1 in modulating the immune response and inflammation through its direct and indirect antiinflammatory mechanisms including lipid transport, high-density lipoprotein (HDL) formation and apoptosis. (C) 2010 The Feinstein Institute for Medical Research, www.feinsteininstitute.org
期刊：
Molecular and Cellular Biology,2013年33(6):1104-1113 ISSN：0270-7306
通讯作者：
Jiang, ZS
作者机构：
[Jiang, Zhi-Sheng] Univ South China, Inst Cardiovasc Dis, Hengyang City, Hunan, Peoples R China.
通讯机构：
[Jiang, ZS ] Univ South China, Inst Cardiovasc Dis, Hengyang City, Hunan, Peoples R China.;Univ South China, Inst Cardiovasc Dis, Hengyang City, Hunan, Peoples R China.
摘要：
LRRC4 is not only a brain-specific gene, but it has also been identified as a tumor suppressor gene for glioma. Promoter methylation of LRRC4 is frequently involved in the inactivation in glioma. MiRNA-mediated gene regulation has recently been demonstrated to play an important role in multiple biological processes related to cancer, including glioma. In this study, we demonstrated that a small regulatory microRNA, hsa-miR-381, an "oncomir", had a major role in glioma progression and that LRRC4 was a target of hsa-miR-381. By regulating LRRC4, hsa-miR-381 increased the in vitro and in vivo proliferation of glioma cells, and this action was associated with decreased inhibition of MEK/ERK and AKT signaling. Conversely, LRRC4, as a glioma suppressor, inhibited the endogenous expression of hsa-miR-381 and decreased cell proliferation and tumor growth. The interaction of hsa-miR-381 and LRRC4 is involved in the pathogenesis of glioma. In addition, the stable expression of hsa-miR-381 in blood provides a novel and promising diagnostic biomarker, and anti-hsa-miR-381 "antagomir" may be an ideal target for glioma therapy. (C) 2011 Elsevier B.V. All rights reserved.
摘要：
Proprotein convertase subtilisin/kexin 9 (PCSK9), a member of the protein-converting enzyme family, is highly expressed in adult hepatocytes and small intestinal enterocytes. To our knowledge, in this study, we demonstrate for the first time that PCSK9 is upregulated in a dose-dependent manner via oxidized low-density lipoprotein (oxLDL) stimulation in THP-1-derived macrophages. PCSK9 small interfering RNA (siRNA) suppresses the oxLDL-induced inflammatory cytokine expression in THP-1-derived macrophages. The exposure of macrophages to oxLDL markedly increased the expression of NF-kappa B protein in the nucleus. However, this effect was significantly attenuated by PCSK9 si RNA. These findings indicate that PCSK9 expression is induced by ox LDL, and that PCSK9 si RNA protects against inflammation via the inhibition of NF-kappa B activation in oxLDL-stimulated THP-1-derived macrophages. Our results suggest that PCSK9 may be used as a therapeutic target for the treatment of atherosclerosis since PCSK9 siRNA suppresses oxLDL-induced I kappa B-alpha degradation and NF-kappa B nuclear translocation into THP-1-derived macrophages.
摘要：
Rationale: Macrophage accumulation of cholesterol leads to foam cell formation which is a major pathological event of atherosclerosis. Recent studies have shown that microRNA (miR)-19b might play an important role in cholesterol metabolism and atherosclerotic diseases. Here, we have identified miR-19b binding to the 3'UTR of ATP-binding cassette transporter A1 (ABCA1) transporters, and further determined the potential roles of this novel interaction in atherogenesis. Objective: To investigate the molecular mechanisms involved in a miR-19b promotion of macrophage cholesterol accumulation and the development of aortic atherosclerosis. Methods and results: We performed bioinformatics analysis using online websites, and found that miR-19b was highly conserved during evolution and directly bound to ABCA1 mRNA with very low binding free energy. Luciferase reporter assay confirmed that miR-19b bound to 3110-3116 sites within ABCA1 3'UTR. MiR-19b directly regulated the expression levels of endogenous ABCA1 in foam cells derived from human THP-1 macrophages and mouse peritoneal macrophages (MPMs) as determined by qRT-PCR and western blot. Cholesterol transport assays revealed that miR-19b dramatically suppressed apolipoprotein AI-mediated ABCA1-dependent cholesterol efflux, resulting in the increased levels of total cholesterol (TC), free cholesterol (FC) and cholesterol ester (CE) as revealed by HPLC. The excretion of H-3-cholesterol originating from cholesterol-laden MPMs into feces was decreased in mice overexpressing miR-19b. Finally, we evaluated the proatherosclerotic role of miR-19b in apolipoprotein E deficient (apoE(-/-)) mice. Treatment with miR-19b precursor reduced plasma high-density lipoprotein (HDL) levels, but increased plasma low-density lipoprotein (LDL) levels. Consistently, miR-19b precursor treatment increased aortic plaque size and lipid content, but reduced collagen content and ABCA1 expression. In contrast, treatment with the inhibitory miR-19b antisense oligonucleotides (ASO) prevented or reversed these effects. Conclusion: MiR-19b promotes macrophage cholesterol accumulation, foam cell formation and aortic atherosclerotic development by targeting ABCA1. (C) 2014 Elsevier Ireland Ltd. All rights reserved.
作者：
Liu, Y. F.;Xiao, Z. Q.;Li, M. X.;Li, M. Y.;Zhang, P. F.;Li, C.;Li, F.;Chen, Y. H.;Yi, H.;Yao, H. X.;Chen, Z-C*
期刊：
The Journal of Pathology,2009年217(1):54-64 ISSN：1096-9896
通讯作者：
Chen, Z-C
作者机构：
[Li, C.; Zhang, P. F.; Yi, H.; Xiao, Z. Q.; Li, F.; Liu, Y. F.; Yao, H. X.; Chen, Z-C; Li, M. X.; Li, M. Y.] Cent S Univ, Xiangya Hosp, Chinese Minist Hlth, Key Lab Canc Proteom, Changsha 410008, Hunan, Peoples R China.;[Li, F.; Liu, Y. F.; Yao, H. X.; Chen, Z-C; Li, M. X.] Cent S Univ, Xiangya Sch Med, Canc Res Inst, Changsha 410078, Hunan, Peoples R China.;[Li, M. X.] Univ S China, Dept Histol & Embryol, Hengyang, Peoples R China.;[Chen, Y. H.] Univ So Calif, Los Angeles, CA USA.;[Chen, Z-C] Cent S Univ, Xiangya Hosp, Chinese Minist Hlth, Key Lab Canc Proteom, 87 Xiangya Rd, Changsha 410008, Hunan, Peoples R China.
通讯机构：
[Chen, Z-C] Cent S Univ, Xiangya Hosp, Chinese Minist Hlth, Key Lab Canc Proteom, 87 Xiangya Rd, Changsha 410008, Hunan, Peoples R China.
摘要：
Metastasis is a common phenomenon and the major lethal cause of lung adenocarcinoma (AdC). To discover novel potential biomarkers associated with lymph node metastasis and prognosis in lung AdC, we assessed differences in protein expression between primary lung AdC with (LNM AdC) and without lymph node metastasis (non-LNM AdC) using a quantitative proteomic approach. Laser capture microdissection was performed to purify the cancer cells from primary lung AdC tissues. The differential proteins between the pooled microdissected non-LNM AdC and LNM AdC tissues were identified by two-dimensional difference gel electrophoresis (2D-DIGE) coupled with mass spectrometry (MS). In this study, twenty proteins were found to be differentially expressed in two types of lung AdC. ANXA3, significantly up-regulated in LNM AdC compared with non-LNM AdC, was validated by western blotting. Immunohistochemistry showed that ANXA3 over-expression was frequently observed in LNM AdCs and matched lymph node metastases compared with non-LNM AdCs. ANXA3 over-expression was significantly associated with advanced clinical stage (p < 0.001) and lymph node metastasis (p < 0.001) and increased relapse rate (p < 0.001) and decreased overall survival (p < 0.001) in lung AdCs. Cox regression analysis indicated ANXA3 over-expression was an independent prognostic factor. Our results indicate that ANXA3 might serve as a novel biomarker for lymph node metastasis and prognosis in lung AdC, and play an important role in lung AdC progression. Copyright (C) 2008 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
