Value of glucose transport protein 1 expression in detecting lymph node metastasis in patients with colorectal cancer

BACKGROUND There are limited data on the use of glucose transport protein 1(GLUT-1)expre-ssion as a biomarker for predicting lymph node metastasis in patients with colorectal cancer.GLUT-1 and GLUT-3,hexokinase(HK)-II,and hypoxia-induced factor(HIF)-1 expressions may be useful biomarkers for detecting primary tumors and lymph node metastasis when combined with fluorodeoxyglucose(FDG)uptake on positron emission tomography/computed tomography(PET/CT).AIM To evaluate GLUT-1,GLUT-3,HK-II,and HIF-1 expressions as biomarkers for detecting primary tumors and lymph node metastasis with 18F-FDG-PET/CT.METHODS This retrospective study included 169 patients with colorectal cancer who underwent colectomy and preoperative 18F-FDG-PET/CT at Chungbuk National University Hospital between January 2009 and May 2012.Two tissue cores from the central and peripheral areas of the tumors were obtained and were examined by a dedicated pathologist,and the expressions of GLUT-1,GLUT-3,HK-II,and HIF-1 were determined using immunohisto-chemical staining.We analyzed the correlations among their expressions,various clinicopathological factors,and the maximum standardized uptake value(SUVmax)of PET/CT.RESULTS GLUT-1 was found at the center or periphery of the tumors in 109(64.5%)of the 169 patients.GLUT-1 positivity was significantly correlated with the SUVmax of the primary tumor and lymph nodes,regardless of the biopsy site(tumor center,P<0.001 and P=0.012;tumor periphery,P=0.030 and P=0.010,respectively).GLUT-1 positivity and negativity were associated with higher and lower sensitivities of PET/CT,respectively,for the detection of lymph node metastasis,regardless of the biopsy site.GLUT3,HK-II,and HIF-1 expressions were not significantly correlated with the SUVmax of the primary tumor and lymph nodes.CONCLUSION GLUT-1 expression was significantly correlated with the SUVmax of 18F-FDG-PET/CT for primary tumors and lymph nodes.Clinicians should consider GLUT-1 expression in preoperative endoscopic biopsy in interpreting PET/CT findings.

GLUT1、GLUT2介导的葡萄糖摄取对小鼠牙齿早期发育的影响

目的:探究葡萄糖转运蛋白(glucose transporter,GLUT)1、GLUT2介导的葡萄糖摄取对小鼠牙齿早期发育的影响。方法:收集胚胎13.5 d(embryonic day 0.5,E13.5)、E14.5、E16.5、E18.5和出生后1 d(postnatal day 1,P1)时期的下颌磨牙牙胚、上颌切牙牙胚;实时荧光定量聚合酶链反应(real time-quantitative polymerase chain reaction,RT-qPCR)和Western blot检测牙胚中Glut1、Glut2 mRNA和蛋白水平;免疫组织化学染色检测牙胚中GLUT1、GLUT2、Ki67、糖原水平。将下颌磨牙牙胚在无糖DMEM培养基、高糖且含不同浓度的根皮素(0、0.25、0.5 mmol/L)的DMEM培养基中培养9 d;并对其进行苏木精-伊红(hematoxylin-eosin,HE)染色。结果:(1)在E13.5时期,GLUT1在成釉器中高表达,细胞增殖活跃,糖原在牙板和牙囊中大量沉积;在E14.5、E18.5时期,GLUT1在成釉器中表达逐渐降低,细胞增殖减少,糖原在成釉器和牙乳头中大量沉积;在P1时期,GLUT1在中间层和内釉上皮中表达较多,细胞增殖增多,糖原在牙乳头中少量沉积。在整个牙胚发育阶段,GLUT2表达相对较少。(2)GLUT1在前成釉细胞、前成牙本质细胞中高表达,而在分化后的成釉细胞、成牙本质细胞中表达较少;GLUT2与GLUT1呈现相反的表达趋势。(3)0.5 mmol/L根皮素能够抑制E13.5时期外植体牙胚发育,0.25 mmol/L根皮素不抑制E13.5、E14.5时期外植体牙胚发育,但能够导致牙胚变小,且具有根皮素浓度依赖性。无糖培养基能够抑制E13.5、E14.5时期外植体牙胚发育。结论:GLUT1、GLUT2在牙齿早期发育中的表达受到精确的时空调控,由GLUT1、GLUT2介导的葡萄糖摄取在小鼠牙齿早期发育中发挥重要作用。

熊果苷调节PI3K/Akt/GLUT1信号通路对胃癌细胞恶性进展的影响

目的 探讨熊果苷(Arb)调节磷脂酰肌醇-3激酶B/蛋白激酶B/葡萄糖转运蛋白1(PI3K/Akt/GLUT1)信号通路对胃癌细胞恶性进展的影响。方法 使用不同浓度(12.5、25、50、100、200、400 mol/L)Arb处理SNU-601细胞,检测细胞活性,筛选最佳浓度;将细胞分为对照组(Control组)、熊果苷低、中、高浓度组(L-Arb组、M-Arb组、H-Arb组)、熊果苷高浓度+PI3K/Akt激活剂组(H-Arb+740 Y-P组),分别检测细胞凋亡率、细胞周期、葡萄糖摄取、乳酸生成、ATP生成、细胞迁移数和细胞侵袭数;Western blot检测Bax、半胱氨酸天冬氨酸蛋白水解酶3(cleaved-caspase3)、B细胞淋巴瘤-2(Bcl-2)、p-PI3K、PI3K、p-Akt、Akt、GLUT1、HKⅡ蛋白表达。结果 12.5~400 mol/L的Arb可显著抑制SNU-601细胞增殖,选择50、100、200 mol/L的Arb进行后续实验。与Control组比较,L-Arb组、M-Arb组、H-Arb组细胞活性、葡萄糖摄取、乳酸生成、ATP生成、细胞迁移数、细胞侵袭数及Bcl-2、p-PI3K/PI3K、p-Akt/Akt、GLUT1、HKⅡ蛋白表达降低,细胞凋亡率和Bax、cleaved-caspase3蛋白表达增加(P<0.05);与H-Arb组比较,H-Arb+740 Y-P组细胞活性、葡萄糖摄取、乳酸生成、ATP生成、细胞迁移数、细胞侵袭数及Bcl-2、p-PI3K/PI3K、p-Akt/Akt、GLUT1、HKⅡ蛋白表达增加,细胞凋亡率和Bax、cleaved-caspase3蛋白表达降低(P<0.05)。结论 Arb通过抑制PI3K/Akt/GLUT1信号通路抑制胃癌细胞恶性进展。

GLUT1在肝癌炎症性贫血患者中的表达及临床意义

目的探讨促葡萄糖转运蛋白1(GLUT1)在肝癌炎症性贫血患者血清中的表达,分析其临床意义。方法收集84例肝癌住院患者,选取其中C-反应蛋白(CRP)高表达的患者72例,根据血红蛋白(Hb)水平分为贫血组和非贫血组。采用酶联免疫吸附试验(ELISA)检测患者血清中GLUT1、铁调素和白细胞介素-6(IL-6)表达水平。分析GLUT1与肝癌炎症性贫血的关系。结果CRP高表达的肝癌患者中贫血发生率为43.06%(31/72)。贫血组血清中GLUT1、CRP、IL-6和铁调素表达水平均高于非贫血组(U分别=323.00、459.00、312.50、336.00,P均<0.05)。GLUT1与Hb呈负相关,与CRP、IL-6和铁调素均呈正相关,差异均有统计学意义(r分别=-0.50、0.56、0.61、0.60,P均<0.05)。结论GLUT1过度表达可能在肝癌患者炎症性贫血中发挥重要作用。

Transforming growth factor beta-1 upregulates glucose transporter 1 and glycolysis through canonical and noncanonical pathways in hepatic stellate cells

BACKGROUND Hepatic stellate cells(HSCs)are the key effector cells mediating the occurrence and development of liver fibrosis,while aerobic glycolysis is an important metabolic characteristic of HSC activation.Transforming growth factor-β1(TGF-β1)induces aerobic glycolysis and is a driving factor for metabolic reprogramming.The occurrence of glycolysis depends on a high glucose uptake level.Glucose transporter 1(GLUT1)is the most widely distributed glucose transporter in the body and mainly participates in the regulation of carbohydrate metabolism,thus affecting cell proliferation and growth.However,little is known about the relationship between TGF-β1 and GLUT1 in the process of liver fibrosis and the molecular mechanism underlying the promotion of aerobic glycolysis in HSCs.AIM To investigate the mechanisms of action of GLUT1,TGF-β1 and aerobic glycolysis in the process of HSC activation during liver fibrosis.METHODS Immunohistochemical staining and immunofluorescence assays were used to examine GLUT1 expression in fibrotic liver tissue.A Seahorse extracellular flux(XF)analyzer was used to examine changes in aerobic glycolytic flux,lactate production levels and glucose consumption levels in HSCs upon TGF-β1 stimulation.The mechanism by which TGF-β1 induces GLUT1 protein expression in HSCs was further explored by inhibiting/promoting the TGF-β1/mothersagainst-decapentaplegic-homolog 2/3(Smad2/3)signaling pathway and inhibiting the p38 and phosphoinositide 3-kinase(PI3K)/AKT signaling pathways.In addition,GLUT1 expression was silenced to observe changes in the growth and proliferation of HSCs.Finally,a GLUT1 inhibitor was used to verify the in vivo effects of GLUT1 on a mouse model of liver fibrosis.RESULTS GLUT1 protein expression was increased in both mouse and human fibrotic liver tissues.In addition,immunofluorescence staining revealed colocalization of GLUT1 and alpha-smooth muscle actin proteins,indicating that GLUT1 expression was related to the development of liver fibrosis.TGF-β1 caused an increase in aerobic glycolysis in HSCs and induced GLUT1 expression in HSCs by activating the Smad,p38 MAPK and P13K/AKT signaling pathways.The p38 MAPK and Smad pathways synergistically affected the induction of GLUT1 expression.GLUT1 inhibition eliminated the effect of TGF-β1 on HSC proliferation and migration.A GLUT1 inhibitor was administered in a mouse model of liver fibrosis,and GLUT1 inhibition reduced the degree of liver inflammation and liver fibrosis.CONCLUSION TGF-β1 induces GLUT1 expression in HSCs,a process related to liver fibrosis progression.In vitro experiments revealed that TGF-β1-induced GLUT1 expression might be one of the mechanisms mediating the metabolic reprogramming of HSCs.In addition,in vivo experiments also indicated that the GLUT1 protein promotes the occurrence and development of liver fibrosis.

GLUT-1在三阴性乳腺癌中的表达及其与临床病理特征和预后的关系研究

目的分析三阴性乳腺癌(TNBC)中葡萄糖转运蛋白1(GLUT-1)的表达水平,并探讨其与患者临床病理特征和预后的关系。方法回顾性选取2019年1月至2021年1月在嘉兴市第一医院行乳腺癌根治术的TNBC患者79例(TNBC组),另择同期本院乳腺良性疾病(包括乳腺炎、乳腺纤维瘤等)患者40例为对照(对照组)。留取TNBC组患者的肿瘤组织与癌旁组织,对照组患者乳腺活检组织。采用qRT-PCR法检测组织标本GLUT-1 mRNA表达情况。收集TNBC组患者的临床病理资料,术后每3个月随访1次,随访时间截至2023年12月,其中随访1年时出现死亡、肿瘤复发进展、出现严重并发症(恶病质)等定义为预后不良。比较TNBC组肿瘤组织、癌旁组织与对照组乳腺活检组织GLUT-1 mRNA表达水平;比较不同临床病理特征的TNBC组患者肿瘤组织GLUT-1 mRNA表达水平;分析不同临床病理特征对TNBC患者术后1年预后不良的预测效能、TNBC患者死亡的危险因素及不同GLUT-1 mRNA表达水平TNBC患者生存情况。结果TNBC组肿瘤组织GLUT-1 mRNA表达水平高于癌旁组织、对照组乳腺活检组织(均P<0.05),而TNBC组癌旁组织与对照组乳腺活检组织GLUT-1 mRNA表达水平比较差异无统计学意义(P>0.05)。低分化、肿瘤直径≥5 cm、Ki-67指数≥70%、有神经脉管浸润、有淋巴结转移、有远处转移及TNM分期Ⅲ期的TNBC组患者肿瘤组织GLUT-1 mRNA表达水平分别高于中-高分化、肿瘤直径<5 cm、Ki-67指数<70%、无神经脉管浸润、无淋巴结转移、无远处转移及TNM分期Ⅰ~Ⅱ期的患者(均P<0.05)。肿瘤组织GLUT-1 mRNA表达水平预测TNBC患者术后1年预后不良的灵敏度、特异度及AUC分别为0.845、0.812、0.842,最佳截断值为0.83,有较高的预测效能(P<0.05)。多因素Cox回归分析显示,GLUT-1 mRNA表达水平≥0.83是TNBC患者死亡的独立危险因素(P<0.05)。肿瘤组织GLUT-1 mRNA表达水平≥0.83的TNBC患者生存期低于<0.83的患者[(27.9±4.6)个月比(36.5±5.8)个月,P<0.05]。结论TNBC患者肿瘤组织GLUT-1表达上调,其与临床病理特征及预后存在密切关系,或可作为TNBC病情及预后评估的标志物。

Effects of suppressing glucose transporter-1 by an antisense oligodeoxynucleotide on the growth of human hepatocellular carcinoma cells

BACKGROUND:The glucose transporter-1(Glut-1),a key ratelimiting factor in the transport and metabolism of glucose in cancer cells,is over-expressed in many human cancer cells and this overexpression is correlated with poor biological behavior. The increased levels of Glut-1 expression in hepatocellular carcinoma(HCC)cells functionally affect tumorigenicity.This study was undertaken to investigate effects of suppressing Glut-1 by an antisense oligodeoxynucleotide(AS-ODN)on the growth of human hepatocellular carcinoma(HepG-2)cells. METHODS:We used AS-ODN targeting against the Glut-1 gene in a HepG-2 cell line.There were four experimental groups: empty pcDNA3.1 vector(mock transfection),pcDNA3.1-anti-Glut(+),pcDNA3.1-Glut(+),and non-transfected HepG-2 cells. The Glut-1 mRNA expression was detected by RT-PCR and the Glut-1 protein expression by Western blotting after cell culture, and the glucose uptake was detected after glucose stimulation in each group. RESULTS:Compared with non-transfected HepG-2 or Glut-1 pcDNA3.1,a down-regulation of Glut-1 mRNA in HepG-2 cells transfected with anti-Glut-1 pcDNA3.1 was noted(P<0.05).Glut-1 protein in HepG-2 cells transfected with Glut-1 AS-ODN was decreased compared with non-transfected HepG-2,Glut-1 pcDNA3.1,or empty vectors. Glucose uptake by the HepG-2 cells transfected with AS-ODN was decreased at 1 hour after glucose stimulation.CONCLUSIONS:The application of Glut-1 AS-ODN can down-regulate the expression of Glut-1 at mRNA and protein,and inhibit glucose uptake partially in HepG-2 cells.The Glut-1 gene maybe a potential therapeutic target for HCC.

Transcriptional activation of glucose transporter 1 in orthodontic tooth movement-associated mechanical response

The interplay between mechanoresponses and a broad range of fundamental biological processes, such as cell cycle progression,growth and differentiation, has been extensively investigated. However, metabolic regulation in mechanobiology remains largely unexplored. Here, we identified glucose transporter 1(GLUT1)—the primary glucose transporter in various cells—as a novel mechanosensitive gene in orthodontic tooth movement(OTM). Using an in vivo rat OTM model, we demonstrated the specific induction of Glut1 proteins on the compressive side of a physically strained periodontal ligament. This transcriptional activation could be recapitulated in in vitro cultured human periodontal ligament cells(PDLCs), showing a time-and dose-dependent mechanoresponse. Importantly, application of GLUT1 specific inhibitor WZB117 greatly suppressed the efficiency of orthodontic tooth movement in a mouse OTM model, and this reduction was associated with a decline in osteoclastic activities. A mechanistic study suggested that GLUT1 inhibition affected the receptor activator for nuclear factor-κ B Ligand(RANKL)/osteoprotegerin(OPG)system by impairing compressive force-mediated RANKL upregulation. Consistently, pretreatment of PDLCs with WZB117 severely impeded the osteoclastic differentiation of co-cultured RAW264.7 cells. Further biochemical analysis indicated mutual regulation between GLUT1 and the MEK/ERK cascade to relay potential communication between glucose uptake and mechanical stress response. Together, these cross-species experiments revealed the transcriptional activation of GLUT1 as a novel and conserved linkage between metabolism and bone remodelling.

Constructing recombinant replication-defective adenoviral vectors that express glucose transporter-1 through in vitro ligation

BACKGROUND： We constructed a homologous recombination bacterial method based on the pAdEasy system, a widely used system, for generating recombinant adenoviral vectors that express glucose transporter- 1 （GLUT 1） in rats, OBJECTIVE： This study was designed to investigate the feasibility of generating recombinant replication-defective adenoviral vectors that express GLUT1 in rats by in vitro ligation based on the Adeno-X^TM system. DESIGN： An in vitro cell-based experiment. SETTING： This study was performed at the Linbaixin Medical Research Center of the Second Hospital Affiliated to Sun Yat-sen University and Central Laboratory for Prevention and Treatment of Tumor, Sun Yat-sen University between January and August 2004. MATERIALS： Male, adult, Sprague Dawley rats were used to extract total RNA from brain tissue. E. coli DH5 a and human embryonic kidney 293 cells （HEK293 cells） used in the present study were cryo-preserved by the Second Hospital Affiliated to Sun Yat-sen University. Rabbit anti-rat GLUT1 polyclonal antibody （Chemicon, U.S.A.） and primers （Shanghai Boya Bioengineering Co., Ltd） were also used. METHODS： E1/E3-deleted replication-defective adenoviral vectors were used. Using in vitro ligation, the target gene was first sub-cloned into a shuttle vector plasmid to obtain the fragment containing target gene expression cassettes by enzyme digestion. Subsequently, the fragment was co-transformed with linearized adenoviral backbone vector into the E. coli strain. The recombinant adenoviral plasmid was transfected into HEK293 cells to assembly recombinant adenoviral vectors with replication capabilities. The procedure was repeated several times for recombinant adenoviral vectors amplification. MAIN OUTCOME MEASURES： Efficiency of recombinant adenoviral vectors to express the target gene was measured by gene and protein expression through polymerase chain reaction and Western Blot assays, respectively. RESULTS： Results demonstrated that recombinant adenoviral vectors successfully expressed GLUT1 protein, with a relative molecular mass of 55000 in HEK293 cells. These results suggest that recombinant adenoviral vectors obtained by homologous bacterial recombination feature high efficiency, rapidness, and simplicity. CONCLUSION： We successfully amplified the rat GLUT1 gene and constructed replication-defective adenoviral vectors expressing GLUT1. The replication-defective adenoviral vectors proved to successfully express the target gene in HEK293 cells.

肾上腺嗜铬细胞瘤患者组织葡萄糖转运蛋白1和拓扑异构酶ⅡA表达与临床特征及预后的相关性分析

目的探讨葡萄糖转运蛋白1(GLUT1)和拓扑异构酶ⅡA(TOP2A)在肾上腺嗜铬细胞瘤(PPGL)患者组织中的表达与临床特征及预后的相关性。方法选取2014年3月至2020年6月于新疆维吾尔自治区人民医院行手术切除的120例PPGL组织作为实验组。另收集同期行手术切除的100例无功能肾上腺腺瘤的肾上腺组织作为对照组。采用免疫组织化学染色法检测组织中GLUT1和TOP2A表达情况。通过Cox回归分析PPGL患者预后的影响因素。结果实验组中GLUT1的低表达率和TOP2A的高表达率均高于对照组[51.7%(62/120)比14.0%(14/100)、52.5%(63/120)比19.0%(19/100)],差异均有统计学意义(χ^(2)=34.225、31.829,均P<0.001)。GLUT1、TOP2A表达水平与镜下组织浸润和Ki-67有关(均P<0.05)。预后良好组中GLUT1高表达和TOP2A低表达比例均高于预后不良组,差异均有统计学意义(均P<0.001)。单因素分析结果显示,患者镜下组织浸润、Ki-67、GLUT1、TOP2A均是影响预后的因素(均P<0.001)。Cox多因素回归分析显示TOP2A、镜下组织浸润及Ki-67均是导致PPGL患者预后不良的危险因素,GLUT1为保护因素(均P<0.001)。结论GLUT1在PPGL组织中呈低表达,TOP2A呈高表达,与患者预后不良有关。

尿酸对老年膝骨关节炎病人软骨GLUT9、URAT1表达的影响

目的探讨老年膝骨关节炎病人血清UA水平与软骨组织中UA转运蛋白的相关性。方法收集2018~2019年就诊于河北省人民医院骨科的老年膝骨关节炎(KOA)病人30例。以所有入选者UA中位数水平(254.6μmol/L)将病人分为血清UA<254.6μmol/L组(A组)和UA≥254.6μmol/L组(B组),检测并比较2组软骨组织中葡萄糖易化转运蛋白9(GLUT9)和尿酸盐转运蛋白1(URAT1)的mRNA和蛋白的表达水平。结果与B组相比,A组病人的体质量、BMI较小,VAS评分较低,差异均有统计学意义(P<0.05)。Pearson相关分析显示,VAS评分与UA水平呈正相关(r=0.396,P<0.05)。B组病人关节软骨中GLUT9、URAT1的mRNA和蛋白表达水平均较A组明显升高,差异有统计学意义(P<0.05)。结论UA水平的升高可能通过增加软骨中GLUT9及URAT1蛋白的表达,使软骨细胞内UA水平升高,诱导软骨细胞氧化损伤,从而参与KOA的发病。

妊娠期肝内胆汁淤积症孕妇血清PDCD4,GLUT1表达水平及其与妊娠结局的相关性研究

目的检测妊娠期肝内胆汁淤积症(intrahepatic cholestasis of pregnancy,ICP)孕妇血清程序性细胞死亡因子4(programmed cell death factor 4,PDCD4)和葡萄糖转运蛋白1(glucose transporter 1,GLUT1)的表达水平,分析二者与孕妇妊娠结局的相关性。方法收集巴中市巴州区妇幼保健院产科2018年7月~2020年7月收治的ICP孕妇126例作为研究组,其中轻度ICP组46例,重度ICP组80例,选择同期该院120例健康产检孕妇作为对照组。采用实时荧光定量PCR(qRT-PCR)法测定ICP孕妇血清PDCD4和GLUT1水平,多因素Logistic回归分析影响ICP孕妇妊娠结局的因素,Pearson相关性分析ICP孕妇血清PDCD4和GLUT1水平的相关性。结果与对照组比较,研究组PDCD4(1.36±0.23 vs 1.02±0.21),GLUT1(1.40±0.22 vs 0.99±0.18)水平升高,差异具有统计学意义(t=15.935,12.090,均P=0.000)。重度ICP组PDCD4(1.41±0.25),GLUT1(1.45±0.22)水平显著高于轻度ICP组(1.27±0.20,1.31±0.21),差异具有统计学意义(t=3.246,3.496,均P<0.05)。研究组羊水胎粪污染(20.63%)、自发性早产(7.14%)、产后出血(8.73%)、宫内窘迫(11.90%)等不良妊娠结局的发生率均高于对照组(0.00%,0.83%,0.83%,1.67%),差异均具有统计学意义(χ^(2)=1.049~29.159,均P<0.05);妊娠结局良好组和妊娠结局不良组患者的发病程度(OR=1.109,95%CI=1.035~1.188)、PDCD4(OR=1.428,95%CI=1.013~2.012)以及GLUT1(OR=1.453,95%CI=1.066~1.980)水平差异具有统计学意义(均P<0.05);多因素Logistic回归分析显示,发病程度、PDCD4,GLUT1为ICP孕妇妊娠结局不良的影响因素(Waldχ^(2)==8.738,1.428,1.453;P=0.003,0.041,0.018);Pearson相关性分析显示,ICP孕妇血清PDCD4与GLUT1水平呈正相关(r=0.460,P<0.05)。结论PDCD4,GLUT1在ICP孕妇血清中表达均上调,二者呈正相关,是ICP孕妇妊娠结局不良的影响因素。

Chlorogenic Acid Maintains Glucose Homeostasis through Modulating the Expression of SGLT-1,GLUT-2,and PLG in Different Intestinal Segments of Sprague-Dawley Rats Fed a High-Fat Diet

Objective To reveal the effects and related mechanisms of chlorogenic acid(CGA)on intestinal glucose homeostasis.Methods Forty male Sprague-Dawley rats were randomly and equally divided into four groups:normal chow(NC),high-fat diet(HFD),HFD with low-dose CGA(20 mg/kg,HFD-LC),and HFD with high-dose CGA(90 mg/kg,HFD-HC).The oral glucose tolerance test was performed,and fast serum insulin(FSI)was detected using an enzyme-linked immunosorbent assay.The m RNA expression levels of glucose transporters(Sglt-1 and Glut-2)and proglucagon(Plg)in different intestinal segments(the duodenum,jejunum,ileum,and colon)were analyzed using quantitative real-time polymerase chain reaction.SGLT-1 protein and the morphology of epithelial cells in the duodenum and jejunum was localized by using immunofluorescence.Results At both doses,CGA ameliorated the HFD-induced body weight gain,maintained FSI,and increased postprandial 30-min glucagon-like peptide 1 secretion.High-dose CGA inhibited the HFD-induced elevation in Sglt-1 expression.Both CGA doses normalized the HFD-induced downregulation of Glut-2 and elevated the expression of Plg in all four intestinal segments.Conclusion An HFD can cause a glucose metabolism disorder in the rat intestine and affect body glucose homeostasis.CGA can modify intestinal glucose metabolism by regulating the expression of intestinal glucose transporters and Plg,thereby controlling the levels of blood glucose and insulin to maintain glucose homeostasis.

GLUT-1在梗阻性结肠癌支架植入术后的表达及临床意义

目的探讨梗阻性结肠癌支架植入术后肿瘤组织中葡萄糖转运蛋白1(GLUT-1)的表达与临床病理特征及无病生存(DFS)和预后的关系。方法收集我院2011—2020年梗阻性结肠癌行支架植入术后且临床资料完整的病例60例。运用免疫组化方法检测GLUT-1在肿瘤组织中的表达,分析其与临床病理特征之间的关系及生存预后的相关性。结果GLUT-1阳性表达率为86.7%,免疫组化评分为0分8例(13.3%),1分7例(11.7%),2分26例(43.3%),3分19例(31.7%)。Kaplan-Meier生存分析显示,GLUT-1表达0分的患者3年DFS为100%,GLUT-1表达1分的患者3年DFS为85.7%,GLUT-1表达2分的患者3年DFS为80.8%,GLUT-1表达3分的患者3年DFS为42.1%。表达3分的患者DFS低于0~2分的患者,差异有统计学意义(P=0.005)。Cox分析显示GLUT-1表达是患者独立的预后危险因素(P=0.001)。结论梗阻性结肠癌支架植入术后GLUT-1的高表达与患者预后相关,可以作为患者术后加强辅助治疗以及密切随访的有效依据。

BcSDR1 is involved in regulation of glucose transport and cAMP and MAPK signaling pathways in Botrytis cinerea

Botrytis cinerea is a typical necrotrophic pathogenic fungus that causes severe diseases in a wide range of plant species, leading to significant economic losses. Our previous study showed that BcSDR1 positively regulates growth,development, and pathogenicity of B. cinerea. However, the regulation mechanism of BcSDR1 and the relationship between BcSDR1 and cAMP and MAPK signaling pathways are not well understood. In this study, transcriptome data showed that BcSDR1 is involved in glucose transmembrane transport, signal transduction, secondary metabolism, and other biological processes. BcSDR1 mutant(BCt41) showed remarkably weak sensitivity to cAMP and MAPK signaling pathways specific inhibitors, SQ22536 and U0126, and significantly decreased cAMP content. The key genes of cAMP and MAPK signaling pathways, BcGB1, BcBTP1, BcBOS1, BcRAS1, and BcBMP3 were significantly upregulated,whereas BcPLC1, BcBCG1, BcCDC4, BcSAK1, BcATF1, and BcBAP1 were significantly downregulated(P<0.05).BcSDR1 was obviously upregulated in BcBCG2, BcBCG3, BcPKA1, and BcPKAR RNA interference(RNAi) mutants, but significantly downregulated in BcPKA2, BcBMP1, and BcBMP3 RNAi mutants. Thus, BcBCG2, BcBCG3, BcPKA1, and BcPKAR negatively regulate BcSDR1 expression, whereas BcPKA2, BcBMP1, and BcBMP3 positively regulate BcSDR1expression.

Photoactivation of GLUT4 translocation promotes glucose uptake via PI3-K/Akt2 signaling in 3T3-L1 adipocytes

Insulin resistance is a hallmark of the metabolic syndrome and type 2 diabetes.Dysfunction of PI-3K/Akt signaling was involved in insulin resistance.Glucose transporter 4(GLUT4)is a keyfactor for glucose uptake in muscle and adipose tissues,which is closely regulated by Pi-3K/Aktsignaling in response to insulin treatment.Low-power laser irradiation(LPLI)has been shown toregulate various physiological processes and induce the synthesis or release of multiple moleculessuch as growth factors,which(especially red and near infrared light)is mainly through theactivation of mitochondrial respiratory chain and the initiation of intracellular signaling path-ways.Nevertheless,it is unclear whether LPLI could promote glucose uptake through activationof PI-3K/Akt/GLUT4 signaling in 3T3L-1 adipocytes.In this study,we investigated how LPLIpromoted glucose uptake through activation of PI-3K/Akt/GLUT4 signaling path way.Here,we showed that GLUT4 was localized to the Golgi apparatus and translocated from cytoplasm tocytomembrane upon LPLI treatment in 3T3L-1 adipocytes,which enhanced glucose uptake.Moreover,we found that glucose uptake was mediated by the PI3-K/Akt2 signaling,but notAkt1 upon LPLI treatment with Akt isoforms gene silence and PI3-K/Akt inhibitors.Collec-tively,our results indicate that PI3-K/Akt2/GLUT4 signaling act as the key regulators forimprovement of glucose uptake under LPLI treatment in 3T3L-i adipocytes.More importantly,our findings suggest that activation of PI3-K/Akt2/GLUT4 signaling by LPLI may provideguidance in practical applications for promotion of glucose uptake in insulin-resistant adiposetissue.

RP11-386G11.10在三阴性乳腺癌组织中的表达及其对细胞增殖和迁移的影响

目的:探讨三阴性乳腺癌组织中长链非编码RNA(lncRNA)RP11-386G11.10的表达及其通过调控miR-1299-3p/葡萄糖转运蛋白-1(GLUT-1)分子轴对三阴性乳腺癌细胞增殖和迁移的影响。方法:收集46例三阴性乳腺癌组织及癌旁组织,RT-qPCR检测RP11-386G11.10在其中的表达以及在正常乳腺上皮MCF-10A细胞和三阴性乳腺癌细胞系(MDA-MB-435、CAL-51、BT-549、MDA-MB-231)中的表达。si-NC慢病毒和si-RP11-386G11.10慢病毒感染BT-549细胞(即si-NC组和si-RP11-386G11.10组),克隆形成实验和细胞划痕实验分别检测细胞增殖和迁移能力;RT-qPCR检测感染后BT-549细胞中miR-1299-3p和GLUT-1 mRNA的表达;双荧光素酶报告基因实验验证RP11-386G11.10与miR-1299-3p的靶向关系。RT-qPCR检测GLUT-1 mRNA在三阴性乳腺癌组织中的表达;Pearson法检测RP11-386G11.10与GLUT-1 mRNA在三阴性乳腺癌组织中表达的关系。免疫印迹检测沉默RP11-386G11.10对BT-549细胞中GLUT-1蛋白以及JAK2/STAT3通路蛋白表达的影响。结果:与癌旁组织相比,三阴性乳腺癌组织中RP11-386G11.10的表达显著升高。与MCF-10A细胞相比,三阴性乳腺癌细胞系(MDA-MB-435、CAL-51、BT-549、MDA-MB-231)中RP11-386G11.10的表达均显著升高。与si-NC组BT-549细胞相比,si-RP11-386G11.10组细胞增殖能力和迁移能力均显著降低,miR-1299-3p表达显著升高,GLUT-1mRNA表达显著降低。RP11-386G11.10能够靶向互补结合miR-1299-3p。与癌旁组织相比,三阴性乳腺癌组织中GLUT-1 mRNA表达显著增加;Pearson法分析显示RP11-386G11.10与GLUT-1 mRNA在三阴性乳腺癌组织中的表达呈正相关。与si-NC组BT-549细胞比较,si-RP11-386G11.10组细胞中GLUT-1蛋白表达显著降低,JAK2/STAT3通路转导被抑制。结论:RP11-386G11.10在三阴性乳腺癌中表达显著上调,沉默RP11-386G11.10能够抑制三阴性乳腺癌BT-549细胞的增殖能力和迁移能力,其作用机制可能与靶向调控miR-1299-3p/GLUT-1分子轴有关。

Effect of exercise during pregnancy on offspring development through ameliorating high glucose and hypoxia in gestational diabetes mellitus

BACKGROUND Gestational diabetes mellitus(GDM)women require prenatal care to minimize short-and long-term complications.The mechanism by which exercise during pregnancy affects organ development and whether glucose transporter(GLUT)1 plays a role in GDM offspring organ development remains unknown.AIM To determine the effect of exercise during pregnancy on the cardiac,hepatic and renal development of GDM mother’s offspring.METHODS Placenta samples were collected from humans and mice.GDM mouse models were created using streptozotocin along with a GDM with exercise group.The hearts,livers and kidneys of 3-and 8-week-old offspring were collected for body composition analysis and staining.The effects of high glucose levels and hypoxia were investigated using HTR8/SVneo.Transwell and wound-healing assays were performed to assess cell migration.Immunofluorescence accompanied with TUNEL and Ki67 staining was used to explore apoptosis and proliferation.RESULTS Exercise during pregnancy downregulated the GLUT1 and hypoxia inducible factor-1αexpression in placenta from individuals with GDM.Cobalt chloride induced hypoxia and high glucose levels also significantly decreased migration and apoptosis of HTR8/SVneo cells.In addition,exercise reduced inflammatory cell infiltration in the liver and decreased the tubular vacuolar area in the kidneys of offspring.CONCLUSION GDM affects the growth and development of organs in offspring.Exercise during pregnancy can reverse adverse effects of GDM on the development of the heart,liver,and kidney in offspring.

血清生长激素释放肽及葡萄糖转运蛋白-1 mRNA对重度子痫前期的预测价值

目的 探讨孕产妇血清生长激素释放肽(Ghrelin)及葡萄糖转运蛋白-1(GTUT-1)mRNA对子痫前期(PE)的预测价值。方法 以2019年1月至2023年1月该院收治的267例经临床诊断为PE的孕产妇作为研究对象,根据病情严重程度将其分为轻度RE组(152例)和重度PE组(115例)。另选取同期在该院产检的94例健康孕产妇作为对照组。比较对照组、轻度PE组及重度PE组孕产妇血清Ghrelin及GTUT-1水平,采用多元Logistic回归分析PE病情严重程度的影响因素,受试者工作特征(ROC)曲线分析Ghrelin及GTUT-1对PE的预测价值。结果 对照组、轻度PE组及重度PE组血清Ghrelin及GTUT-1 mRNA依次升高,组间比较差异均有统计学意义(P<0.05);多元Logistic回归分析显示,UA、Ghrelin、GLUT-1 mRNA、CRP、IL-6为重度PE为重度PE的独立危险因素(P<0.05);ROC曲线分析显示,Ghrelin和GLUT-1 mRNA预测重度PE的cut-off值分别为19.56μg/L、1.11,对应曲线下面积(AUC)分别为为0.756、0.769。结论 孕产妇血清Ghrelin及GTUT-1 mRNA对重度PE有一定预测价值。

miR-340对脑胶质瘤中GLUT1/6表达及细胞增殖、迁移的影响

目的探究脑胶质瘤微小核糖核酸microRNA-340(miR-340)与IDH1、P53、CD34、Ki-67表达的关系以及初步探索miR-340对胶质瘤细胞功能和葡萄糖转运蛋白1/6(glucose transporter 1/6,GLUT1/6)表达的影响.方法采用实时荧光定量PCR(qRT-PCR)法检测脑胶质瘤组织标本及胶质瘤细胞系miR-340的表达水平;细胞功能试验检测miR-340对胶质瘤的影响;qRT-PCR、蛋白质免疫印迹检测转染miR-340对GLUT1/6表达的影响.结果miR-340在正常脑组织中高表达,胶质瘤组织中低表达(P<0.001);miR-340 mimics可以抑制U87、U251细胞增殖、增加凋亡、减弱迁移侵袭.miR-340 mimics可以显著抑制胶质瘤细胞中GLUT1/6的表达(P<0.001).结论胶质瘤miR-340水平降低,且与Ki-67表达呈明显负相关.miR-340通过降低胶质瘤中GLUT1/6表达从而影响胶质瘤葡萄糖转运以抑制增殖,降低恶性程度.