Clinical implications of forkhead box M1, cyclooxygenase-2, and glucose-regulated protein 78 in breast invasive ductal carcinoma

BACKGROUND Breast infiltrating ductal carcinoma(BIDC)represents the largest heterotypic tumor group,and an in-depth understanding of the pathogenesis of BIDC is key to improving its prognosis.AIM To analyze the expression profiles and clinical implications of forkhead box M1(FOXM1),cyclooxygenase-2(COX-2),and glucose-regulated protein 78(GRP78)in BIDC.METHODS A total of 65 BIDC patients and 70 healthy controls who presented to our hospital between August 2019 and May 2021 were selected for analysis.The peripheral blood FOXM1,COX-2,and GRP78 levels in both groups were measured and the association between their expression profiles in BIDC was examined.Additionally,we investigated the diagnostic value of FOXM1,COX-2,and GRP78 in patients with BIDC and their correlations with clinicopathological features.Furthermore,BIDC patients were followed for 1 year to identify factors influencing patient prognosis.RESULTS The levels of FOXM1,COX-2,and GRP78 were significantly higher in BIDC patients compared to healthy controls(P<0.05),and a positive correlation was observed among them(P<0.05).Receiver operating characteristic analysis demonstrated that FOXM1,COX-2,and GRP78 had excellent diagnostic value in predicting the occurrence of BIDC(P<0.05).Subsequently,we found significant differences in FOXM1,COX-2,and GRP78 levels among patients with different histological grades and metastasis statuses(with vs without)(P<0.05).Cox analysis revealed that FOXM1,COX-2,GRP78,increased histological grade,and the presence of tumor metastasis were independent risk factors for prognostic death in BIDC(P<0.001).CONCLUSION FOXM1,COX-2,and GRP78 exhibit abnormally high expression in BIDC,promoting malignant tumor development and closely correlating with prognosis.These findings hold significant research implications for the future diagnosis and treatment of BIDC.

抑制miR-21-5p表达通过靶向FOXP2调控肺癌细胞增殖和凋亡

目的探究抑制miR-21-5p表达通过靶向叉头蛋白P2抗体(FOXP2)对肺癌细胞增殖和凋亡的影响。方法实时荧光定量聚合酶链反应(qRT-PCR)、Western印迹检测正常肺上皮细胞株及肺癌细胞株中miR-21-5p、FOXP2的表达水平,双荧光素酶实验检测miR-21-5p与FOXP2的联系,Western印迹检测转染后A549细胞中细胞周期蛋白(Cyclin)D1、p21、p27、B细胞淋巴瘤(Bcl)-2、Bcl-2相关x蛋白(Bax)、酶切含半胱氨酸的天冬氨酸蛋白水解酶(caspase)3蛋白水平,噻唑蓝(MTT)法检测细胞增殖情况,流式细胞仪检测细胞凋亡情况。结果与正常肺上皮细胞BEAS-2B比较,肺癌细胞株A549、NCl-H460、H1299中miR-21-5p表达水平均显著升高,FOXP2 mRNA及蛋白水平均显著降低(均P<0.05)。miR-21-5p组FOXP2蛋白表达水平(0.29±0.03)显著低于miR-NC组(0.62±0.06),anti-miR-21-5p组FOXP2蛋白表达水平(0.94±0.08)显著高于anti-miR-NC组(0.61±0.05,均P<0.05)。与anti-miR-NC组比较,anti-miR-21-5p组miR-21-5p水平显著降低,48 h、72 h OD值显著降低,CyclinD1蛋白水平显著降低,p21、p27蛋白水平显著升高,细胞凋亡率显著升高,Bcl-2蛋白水平显著降低,Bax、酶切caspase3蛋白水平显著升高(均P<0.05)。与pcDNA组比较,pcDNA-FOXP2组FOXP2水平显著升高,48 h、72 h OD值显著降低,CyclinD1蛋白水平显著降低,p21蛋白水平显著升高,细胞凋亡率显著升高,Bcl-2蛋白水平显著降低,Bax蛋白水平显著升高(P<0.05)。与anti-miR-21-5p+si-NC组比较,anti-miR-21-5p+si-FOXP2组FOXP2水平显著降低,48 h、72 h OD值显著升高,细胞凋亡率显著降低,CyclinD1、Bcl-2蛋白水平显著升高,p21、Bax蛋白水平显著降低(均P<0.05)。结论miR-21-5p靶向调控肺癌细胞中FOXP2的表达,抑制FOXP2的表达能逆转抑制miR-21-5p引起的细胞增殖抑制,凋亡促进作用。

Foxp2小鼠模型中发育性言语障碍的分子遗传学研究

叉头框P2基因(FOXP2)是第一例发现的与一种特异性言语和语言障碍,即发育性言语失用症相关的基因。这一发现开启了研究相关神经通路的崭新方向。FOXP2在各种脊椎动物中显示了序列和神经系统表达的显著高度保守性,例如它在人胎脑中与对等阶段的小鼠胎脑中表达模式高度相似。Foxp2小鼠模型包括基因敲除鼠,类似KE家族病理突变的鼠以及Foxp2被人源化的鼠。本文将从分子网络、感觉处理、运动技能学习等三方面综述对Foxp2小鼠模型的研究,以阐明语言障碍的遗传基础。

miR-132-3p靶向抑制FOXP2基因表达对乳腺癌细胞增殖、侵袭和迁移的影响

目的观察miR-132-3p对乳腺癌细胞增殖、侵袭和迁移的影响,并探讨其作用机制。方法qRT-PCR检测乳腺癌细胞MCF-7、MDA-MB-231、BT549和BT474及人乳腺上皮细胞HBL-100中miR-132-3p和叉头框蛋白(FOX)P2 mRNA表达,Western印迹检测FOXP2蛋白表达。分别转染miR-con、miR-132-3p mimics和si-con、si-FOXP2构建过表达miR-132-3p或沉默FOXP2的MDA-MB-231细胞系,四甲基噻唑蓝(MTT)检测细胞增殖,Transwell检测细胞迁移和侵袭,Western印迹检测细胞周期依赖性蛋白激酶(CDK)1、细胞周期蛋白(Cyclin)D1、基质金属蛋白酶(MMP)-2和MMP-9蛋白表达。双荧光素酶报告基因实验验证miR-132-3p与FOXP2的靶向关系。结果与HBL-100细胞相比,乳腺癌细胞MCF-7、MDA-MB-231、BT549和BT474中miR-132-3p表达显著降低(P<0.05),FOXP2 mRNA和蛋白表达显著升高(P<0.05)。过表达miR-132-3p或沉默FOXP2均可有效抑制MDA-MB-231细胞的增殖、迁移和侵袭,抑制CDK1、Cyclin D1、MMP-2和MMP-9蛋白表达。miR-132-3p靶向负调控FOXP2表达,过表达FOXP2可部分逆转miR-132-3p过表达对MDA-MB-231细胞增殖、迁移和侵袭能力的抑制作用及相关蛋白的调控作用。结论miR-132-3p过表达可抑制乳腺癌细胞的增殖、迁移和侵袭,其机制可能与miR-132-3p负调控FOXP2蛋白表达有关。

MiR-19a-3p regulates the Forkhead box F2-mediated Wnt/β-catenin signaling pathway and affects the biological functions of colorectal cancer cells

BACKGROUND Colorectal cancer(CRC)is one of the most common malignancies worldwide.AIM To explore the expression of microRNA miR-19a-3p and Forkhead box F2(FOXF2)in patients with CRC and the relevant mechanisms.METHODS Sixty-two CRC patients admitted to the hospital were enrolled into the study group,and sixty healthy people from the same period were assigned to the control group.Elbow venous blood was sampled from the patients and healthy individuals,and blood serum was saved for later analysis.MiR-19a-3p mimics,miR-19a-3p inhibitor,miR-negative control,small interfering-FOXF2,and short hairpin-FOXF2 were transfected into HT29 and HCT116 cells.Then quantitative polymerase chain reaction was performed to quantify the expression of miR-19a-3p and FOXF2 in HT29 and HCT116 cells,and western blot(WB)analysis was conducted to evaluate the levels of FOXF2,glycogen synthase kinase 3 beta(GSK-3β),phosphorylated GSK-3β(p-GSK-3β),β-catenin,p-β-catenin,α-catenin,Ncadherin,E-cadherin,and vimentin.The MTT,Transwell,and wound healing assays were applied to analyze cell proliferation,invasion,and migration,respectively,and the dual luciferase reporter assay was used to determine the correlation of miR-19a-3p with FOXF2.RESULTS The patients showed high serum levels of miR-19a-3p and low levels of FOXF2,and the area under the curves of miR-19a-3p and FOXF2 were larger than 0.8.MiR-19a-3p and FOXF2 were related to sex,tumor size,age,tumor-nodemetastasis staging,lymph node metastasis,and differentiation of CRC patients.Silencing of miR-19a-3p and overexpression of FOXF2 suppressed the epithelialmesenchymal transition,invasion,migration,and proliferation of cells.WB analysis revealed that silencing of miR-19a-3p and FOXF2 overexpression significantly suppressed the expression of p-GSK-3β,β-catenin,N-cadherin,and vimentin;and increased the levels of GSK-3β,p-β-catenin,α-catenin,and Ecadherin.The dual luciferase reporter assay confirmed that there was a targeted correlation of miR-19a-3p with FOXF2.In addition,a rescue experiment revealed that there were no differences in cell proliferation,invasion,and migration in HT29 and HCT116 cells co-transfected with miR-19a-3p-mimics+sh-FOXF2 and miR-19a-3p-inhibitor+si-FOXF2 compared to the miR-negative control group.CONCLUSION Inhibiting miR-19a-3p expression can upregulate the FOXF2-mediated Wnt/β-catenin signaling pathway,thereby affecting the epithelial-mesenchymal transition,proliferation,invasion,and migration of cells.Thus,miR-19a-3p is likely to be a therapeutic target in CRC.

Forkhead box protein A2 and T helper type 2-mediated pulmonary inflammation

The transcription factor forkhead box protein A2(FOXA2, also known as hepatocyte nuclear factor 3β or transcription factor 3β), has been found to play pivotal roles in multiple phases of mammalian life, from the early development to the organofaction, and subsequently in homeostasis and metabolism in the adult. In the embryonic development period, FOXA2 is require d for the formation of the primitive node and notochord, and its absence results in embryonic lethality. Moreover, FOXA2 plays an important role not only in lung development, but also in T helper type 2(Th2)-mediated pulmonary inflammation and goblet cell hyperplasia. In this article, the role of FOXA2 in lung development and Th2-mediated pulmonary inflammation, as well as in goblet cell hyperplasia, is reviewed. FOXA2 deletion in airway epithelium results into Th2-mediated pulmonary inflammation and goblet cell hyperplasia in developing lung. Leukotriene pathway and signal transducers and activators of transcription 6 pathway may mediate this inflammation through recruitment and activation of denditric cell during lung developments. FOXA2 is a potential treatment target for lung diseases with Th2 inflammation and goblet cell hyperplasia, such as asthma and chronic obstructive pulmonary disease.

Correlation of Forkhead Box c2 with Subtypes and Invasive Ability of Invasive Breast Cancer

Forkhead Box c2 （FOXC2） is a member of forkhead/winged-helix family of transcription factors. The relationship between FOXC2 and invasive breast cancers, including basal-like breast cancer （BLBC, a subtype of breast cancer）, remains to be elucidated. In this study, immunohistochemistry was used to detect the expression of FOXC2 in samples from 103 cases of invasive breast cancers and 15 cases of normal mammary glands. The relationship between FOXC2 and clinical parameters of invasive breast cancers such as patient＇s age, tumor size, lymph node metastasis, tumor grade, the expression of ER, PR, HER-2 and p53, and Ki-67 labeling index （LI） was evaluated. The expression of FOXC2 was detected in parent MCF7 ceils, MCF cells transfected with FOXC2 expression vectors and MDA-MB- 435 cells by immunohistochemistry and Western blotting. Transwell assay was used to determine the invasive ability of these cells. The results showed that FOXC2 was strongly expressed in basal epithelial cells in normal mammary glands and weakly expressed or even not expressed in glandular epithelial cells. The majority of invasive breast cancers （71.8%, 74/103） had negative or weak expression of FOXC2. However, FOXC2 was strongly expressed in 60.7% of BLBCs. Moreover, FOXC2 was related with tumor grade, p53 expression, ki-67 LI and lymph nodes metastasis. It was expressed in FOXC2- transfected MCF cells and MDA-MB-435 cells but not in parent MCF cells. Transwell assay revealed that MCF cells transfected with FOXC2 expression vectors were more aggressive than the parent MCF ceils, suggesting a positive correlation between FOXC2 and the invasion of breast cancer. It was con- cluded that there is a significant association between FOXC2 and the metastasis of invasive breast can- cer. FOXC2 may be used as a new marker for the diagnosis and prognosis prediction of different sub- tvpes of invasive breast cancers.

老年心力衰竭并发肺炎患者血清FOXM1和IGF2表达水平及与预后价值研究

目的探究血清叉头盒蛋白M1(forkhead box protein M1,FOXM1)和胰岛素样生长因子2(insulin-like growth factor 2,IGF2)表达对老年心力衰竭合并肺炎患者预后的预测价值。方法将邯郸市中心医院2021年3月~2022年6月收治的126例老年心力衰竭并发肺炎患者设为病例组,并根据随访情况将122例患者分为预后不良组(n=33)和预后良好组(n=89),另选取该院同期126例健康体检者为对照组。检测两组(病例组和对照组)血清FOXM1和IGF2水平,检测病例组用力肺活量(forced vital capacity,FVC)和第一秒用力呼容积(forced expiratory volume in one second,FEV1)。采用Spearman分析法分析老年心力衰竭并发肺炎患者血清FOXM1和IGF2水平与心功能分级的相关性;受试者工作特征(receiver operating characteristic,ROC)曲线分析血清FOXM1和IGF2水平对老年心力衰竭并发肺炎患者预后的预测价值。结果与对照组比较,病例组血清FOXM1(2.39±0.55 vs 1.06±0.21)和IGF2(71.33±7.96pg/ml vs 47.82±5.14pg/ml)水平明显较高,差异有统计学意义(t=25.358,27.581,均P<0.05);与预后良好组比较,预后不良组血清FOXM1(3.87±1.06 vs 1.95±0.51)和IGF2水平(85.88±9.54pg/ml vs 69.14±8.73pg/ml)明显较高,差异具有统计学意义(t=13.453,9.174,均P<0.05);预后良好组和预后不良组心功能分级比较差异有统计学意义(χ^(2)=7.120,P<0.05),且与预后不良组比较,预后良好组FEV1(1.24±0.32L vs 1.08±0.25L)和FEV1/FVC(55.46%±5.77%vs 52.30%±5.38%)明显较高,差异有统计学意义(t=2.592,2.735,均P<0.05);老年心力衰竭并发肺炎患者血清FOXM1水平和IGF2水平与心功能分级呈显著正相关(r=0.496,0.517,均P<0.05)。ROC曲线结果显示,血清FOXM1单独预测老年心力衰竭并发肺炎患者预后的曲线下面积(area under the curve,AUC)为0.854(95CI%:0.779~0.912),其敏感度、特异度分别为75.76%,86.52%,最佳截断值为2.75;IGF2单独预测老年心力衰竭并发肺炎患者预后的AUC为0.874(95CI%:0.802~0.927),其敏感度、特异度分别为72.73%,85.39%,最佳截断值为78.30 pg/ml;二者联合预测老年心力衰竭并发肺炎患者预后的AUC显著大于血清FOXM1和IGF2单独诊断的AUC(Z=2.413,2.737,P=0.006,0.016)。结论血清FOXM1和IGF2水平在老年心力衰竭并发肺炎患者中升高,且二者联合检测对患者预后具有较高的预测价值。

Slit引导配体2通过调控AMPK/SIRT1-FoxO1信号通路影响糖尿病小鼠视网膜血管损伤的机制研究

目的 探讨Slit引导配体2(SLIT2)是否通过调控腺苷单磷酸活化蛋白激酶(AMPK)/转录沉默信息调节因子1(SIRT1)-叉头盒蛋白O1(FoxO1)信号通路通过对糖尿病小鼠视网膜血管损伤的影响。方法 30只db/db小鼠随机分为DR组(db/db小鼠)、DR+阴性对照载体组(DR+sh-NC组)和DR+sh-SLIT2组,每组10只。另选10只db/m小鼠为对照组。DR+sh-NC组和DR+sh-SLIT2组麻醉后分别在双眼玻璃体腔内注射sh-SLIT2的腺相关病毒(AAV)载体。眼底荧光血管造影(FFA)和苏木精伊红染色观察视网膜血管病变;酶联免疫吸附测定检测血清白细胞介素-6(IL-6),肿瘤坏死因子α(TNF-α)和血管内皮生长因子(VEGF)的水平,荧光定量PCR检测SLIT2 mRNA表达;Western blot检测视网膜组织SLIT2、AMPK、SIRT1、FoxO1蛋白水平。结果 与对照组相比,DR组、DR+sh-NC组、DR+sh-SLIT2组血糖、每日饮水量、每日排尿量、食物摄入量及体质量均明显升高(P<0.05);与对照组相比,DR组视网膜存在血管病变及病理损伤,SLIT2 mRNA及蛋白表达、IL-6、TNF-α和VEGF水平,FoxO1蛋白水平均明显升高(P<0.05),AMPK、SIRT1蛋白水平均明显降低(P<0.05);与DR+sh-NC组相比,DR+sh-SLIT2组的视网膜血管病变及病理损伤明显减轻,SLIT2 mRNA及蛋白表达、IL-6、TNF-α和VEGF水平,FoxO1蛋白水平均明显降低(P<0.05),AMPK、SIRT1蛋白水平均明显升高(P<0.05)。结论 沉默SLIT2表达显著改善糖尿病小鼠视网膜血管损伤及炎症水平,这可能是通过调控AMPK/SIRT1-FoxO1信号通路发挥作用的。

食管癌组织中FOXM1和RNF2表达与临床病理参数及预后的相关性分析

目的探讨食管癌组织中叉头盒蛋白M1(FOXM1)和环指蛋白2(RNF2)表达与临床病理参数及预后的相关性。方法选取我院2018年1月至2019年12月期间收治的120例食管癌患者作为研究对象,收集术中切除的食管癌组织标本及癌旁组织标本。另选取45例食管癌患者并收集术中切除的食管癌组织标本作为验证组。检测食管癌组织及癌旁组织中FOXM1和RNF2的表达水平,分析二者相关性及其与患者预后状态的关系,并探讨影响食管癌患者预后状态的影响因素。结果食管癌组织中FOXM1和RNF2表达水平均高于癌旁组织(P<0.05);食管癌组织FOXM1和RNF2表达水平呈正相关(r=0.625,P<0.05);FOXM1高表达组及RNF2高表达组TNM分期为Ⅲ+Ⅳ期、发生淋巴结转移、浸润深度为深层的食管癌患者所占比例高于FOXM1低表达组及RNF2低表达组(P<0.05);癌组织中呈现出FOXM1低表达及RNF2低表达的食管癌患者其3年生存率(47.45%、48.33%)明显高于FOXM1高表达和RNF2高表达者(27.87%、26.67%)( χ^(2)=4.302、6.009,P=0.038、0.014);TNM分期、淋巴结转移、浸润深度、FOXM1及RNF2是食管癌患者死亡的危险因素(P<0.05);验证组中,FOXM1高表达组和RNF2高表达组发生淋巴结转移、TNM分期为Ⅲ+Ⅳ期、浸润深度为深层的患者所占比例分别高于FOXM1低表达组和RNF2低表达组(P<0.05),而3年生存率则分别低于FOXM1低表达组和RNF2低表达组(P<0.05)。结论FOXM1和RNF2在食管癌患者癌组织中的表达水平与其临床病理特征及3年生存率有关。

Autocrine IL-8 Contributes to Propionibacterium Acnes-induced Proliferation and Differentiation of HaCaT Cells via AKT/FOXO1/Autophagy

Objective Proprionibacterium acnes(P.acnes)-induced inflammatory responses,proliferation and differentiation of keratinocytes contribute to the progression of acne vulgaris(AV).P.acnes was found to enhance the production of interleukin-8(IL-8)by keratinocytes.This study aimed to investigate the role of IL-8 in P.acnes-induced proliferation and differentiation of keratinocytes and the underlying mechanism.Methods The P.acnes-stimulated HaCaT cell(a human keratinocyte cell line)model was established.Western blotting and immunofluorescence were performed to detect the expression of the IL-8 receptors C-X-C motif chemokine receptor 1(CXCR1)and C-X-C motif chemokine receptor 2(CXCR2)on HaCaT cells.Cell counting kit-8(CCK-8)assay,5-ethynyl-20-deoxyuridine(EdU)assay and Western blotting were performed to examine the effects of IL-8/CXCR2 axis on the proliferation and differentiation of HaCaT cells treated with P.acnes,the IL-8 neutralizing antibody,the CXCR2 antagonist(SB225002),or the CXCR1/CXCR2 antagonist(G31P).Western blotting,nuclear and cytoplasmic separation,CCK-8 assay,and EdU assay were employed to determine the downstream pathway of CXCR2 after P.acnes-stimulated HaCaT cells were treated with the CXCR2 antagonist,the protein kinase B(AKT)antagonist(AZD5363),or the constitutively active forkhead box O1(FOXO1)mutant.Finally,autophagy markers were measured in HaCaT cells following the transfection of the FOXO1 mutant or treatment with the autophagy inhibitor 3-methyladenine(3-MA).Results The expression levels of CXCR1 and CXCR2 were significantly increased on the membrane of HaCaT cells following P.acnes stimulation.The IL-8/CXCR2 axis predominantly promoted the proliferation and differentiation of P.acnes-induced HaCaT cells by activating AKT/FOXO1/autophagy signaling.In brief,IL-8 bound to its receptor CXCR2 on the membrane of keratinocytes to activate the AKT/FOXO1 axis.Subsequently,phosphorylated FOXO1 facilitated autophagy to promote the proliferation and differentiation of P.acnes-induced keratinocytes.Conclusion This study demonstrated the novel autocrine effect of IL-8 on the proliferation and differentiation of P.acnes-induced keratinocytes,suggesting a potential therapeutic target for AV.

转录因子HNF1A、HNF4A和FOXA2调节肝细胞蛋白质N-糖基化

Hepatocyte nuclear factor 1 alpha(HNF1A),hepatocyte nuclear factor 4 alpha(HNF4A),and forkhead box protein A2(FOXA2)are key transcription factors that regulate a complex gene network in the liver,cre-ating a regulatory transcriptional loop.The Encode and ChIP-Atlas databases identify the recognition sites of these transcription factors in many glycosyltransferase genes.Our in silico analysis of HNF1A,HNF4A.and FOXA2 binding to the ten candidate glyco-genes studied in this work confirms a significant enrich-ment of these transcription factors specifically in the liver.Our previous studies identified HNF1A as a master regulator of fucosylation,glycan branching,and galactosylation of plasma glycoproteins.Here,we aimed to functionally validate the role of the three transcription factors on downstream glyco-gene transcriptional expression and the possible effect on glycan phenotype.We used the state-of-the-art clus-tered regularly interspaced short palindromic repeats/dead Cas9(CRISPR/dCas9)molecular tool for the downregulation of the HNF1A,HNF4A,and FOXA2 genes in HepG2 cells-a human liver cancer cell line.The results show that the downregulation of all three genes individually and in pairs affects the transcrip-tional activity of many glyco-genes,although downregulation of glyco-genes was not always followed by an unambiguous change in the corresponding glycan structures.The effect is better seen as an overall change in the total HepG2 N-glycome,primarily due to the extension of biantennary glycans.We propose an alternative way to evaluate the N-glycome composition via estimating the overall complexity of the glycome by quantifying the number of monomers in each glycan structure.We also propose a model showing feedback loops with the mutual activation of HNF1A-FOXA2 and HNF4A-FOXA2 affecting glyco-genes and protein glycosylation in HepG2 cells.

基于FOXO3a/Nrf2信号通路探讨尿毒症骨骼肌自噬的分子机制

目的:探讨叉头盒(forkhead box,FOX)O蛋白3a(forkhead box O3a,FOXO3a)/转录因子NF-E2相关因子2(nuclear factor erythroid 2-related factor 2,Nrf2)信号通路的调节尿毒症骨骼肌自噬的机制。方法:24只小鼠随机分为假手术组(Sham组)、慢性肾病(chronic kidney disease,CKD)组和CKD+AST-120(AST)组,每组8只。通过5/6肾切除术建立CKD模型。在肾切除术后4周,CKD+AST组小鼠给予含AST-120粉末(尿毒毒素的木炭吸附剂)饮食,持续8周。透射电子显微镜观察腓肠肌组织中的自噬溶酶体。C2C12小鼠成肌细胞分为对照(Control,Con)组、硫酸吲哚酚(indophenol sulfate,IS)组、IS+si-FOXO3a组。实时RT-PCR和蛋白质印迹法分析FOXO3a表达情况。Giemsa染色对C2C12肌管直径进行量化。结果:与CKD组相比,CKD+AST组血清IS水平和腓肠肌组织中FOXO3a表达、自噬溶酶体的数量明显降低(P<0.05),以及腓肠肌、胫骨前肌、比目鱼肌质量明显增加(P<0.05)。与IS组相比,IS+si-FOXO3a组C2C12细胞中FOXO3a、肌肉萎缩盒基因(Muscle Atrophy F-box,atrogin-1)肌肉环脂蛋白1(muscle RING-finger protein-1,MuRF-1)的表达和细胞自噬通量明显降低(P<0.05),以及细胞直径明显增加(P<0.05)。与Sham组相比,CKD组小鼠腓肠肌组织中Kelch样ECH相关蛋白1(Kelch-like ECH-associated protein 1,KEAP1)表达明显升高(P<0.05),和NRF2表达明显降低(P<0.05)。AST治疗逆转了CKD小鼠腓肠肌组织中这些蛋白的变化。si-NRF2逆转了FOXO3a敲低对自噬流量的抑制作用。结论:IS可能通过持续刺激FOXO3a-KEAP1-NRF2轴促进骨骼肌过度自噬,参与尿毒症少肌症的发病机制。

2型糖尿病周围神经病变患者血清FOXO3a、PDGF表达及临床意义

目的 探究2型糖尿病周围神经病变(DPN)患者血清叉头盒O类转录因子-3a(FOXO3a)、血小板源性生长因子(PDGF)表达及临床意义。方法 选取2020年12月—2022年12月中部战区总医院接收的110例2型糖尿病患者作为研究对象。根据是否并发DPN将患者分为无并发症组(NDPN组)和周围神经病变组(DPN组),分别有45和65例,随机选取同期在该院体检的健康人群30例作为对照组(NC组)。采集受试者的一般资料及血脂、血糖等相关生化指标。采用酶联免疫吸附试验检测3组受试者血清FOXO3a、PDGF水平。采用受试者工作特征(ROC)曲线分析血清FOXO3a、PDGF水平对2型糖尿病发生DPN的预测价值。结果 各组年龄、病程比较,差异均有统计学意义(P <0.05)。各组性别构成、体质量指数、舒张压、收缩压、丙氨酸氨基转移酶、天冬氨酸氨基转移酶、肌酐、尿素氮水平比较,差异均无统计学意义(P>0.05)。NDPN组与DPN组动脉粥样硬化、视网膜病变、降压药使用史、注射胰岛素、服用二甲双胍构成比比较,差异均无统计学意义(P>0.05)。各组总胆固醇、甘油三酯、低密度脂蛋白胆固醇、高密度脂蛋白胆固醇、空腹C肽、踝肱指数比较,差异均无统计学意义(P>0.05)。NDPN组、DPN组糖化血红蛋白、空腹血糖水平较NC组高(P <0.05),DPN组运动神经传导速度、感觉神经传导速度水平较NDPN组低(P <0.05)。NDPN组、DPN组FOXO3a、PDGF水平较NC组高(P <0.05),DPN组FOXO3a、PDGF水平较NDPN组高(P <0.05)。ROC曲线分析结果显示,血清FOXO3a、PDGF以及联合诊断糖尿病患者是否发生DPN的曲线下面积分别为0.695(95%CI:0.590,0.779)、0.636(95%CI:0.539,0.726)、0.732(95%CI:0.639,0.812),敏感性分别为80.00%(95%CI:0.733,0.848)、92.31%(95%CI:0.856,0.977)、89.23%(95%CI:0.838,0.954),特异性分别为55.56%(95%CI:0.438,0.654)、33.33%(95%CI:0.238,0.452)、51.11%(95%CI:0.416,0.597)。结论 2型糖尿病DPN患者体内血清FOXO3a、PDGF水平异常,提示其与2型糖尿病周围神经病变发生、发展有相关性。

莫诺苷调控糖尿病周围神经病变大鼠坐骨神经氧化应激损伤的机制

目的探讨莫诺苷对糖尿病周围神经病变(DPN)大鼠坐骨神经氧化应激损伤的影响。方法48只大鼠分为对照组、DPN组、莫诺苷组、莫诺苷+Brusatol组,每组12只。检测空腹血糖、机械痛阈值、热痛阈值、坐骨神经传导速度的变化;透射电镜观察坐骨神经超微结构变化;ELISA法检测坐骨神经中白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、丙二醛(MDA)含量及超氧化物歧化酶(SOD)活性;Western blot检测坐骨神经中核因子E2相关因子2(Nrf2)、沉默信息调节因子2相关酶1(SIRT_(1))、叉头转录因子o3(Foxo3)蛋白。结果DPN组、莫诺苷组、莫诺苷+Brusatol组坐骨神经髓鞘受损,空腹血糖、热痛阈值、坐骨神经中IL-6、TNF-α、MDA含量高于对照组,机械痛阈值、感觉和运动神经传导速度、坐骨神经中SOD活性及Nrf2、SIRT_(1)、Foxo3蛋白低于对照组,差异有统计学意义(P<0.05);莫诺苷组坐骨神经组织超微结构损伤改善,空腹血糖、热痛阈值、坐骨神经中IL-6、TNF-α、MDA含量低于DPN组,机械痛阈值、感觉和运动神经传导速度、坐骨神经中SOD活性及Nrf2、SIRT_(1)、Foxo3蛋白表达高于DPN组,差异有统计学意义(P<0.05);在莫诺苷组基础上给予Nrf2抑制剂Brusatol后,莫诺苷的保护作用被逆转。结论莫诺苷可能通过激活Nrf2/SIRT_(1)/Foxo3通路改善DPN大鼠坐骨神经氧化应激损伤。

血清组蛋白去乙酰化酶2、叉头框蛋白A1在子痫前期诊断及妊娠结局评估中的价值

目的探究血清组蛋白去乙酰化酶2(HDAC2)、叉头框蛋白A1(FOXA1)表达在子痫前期(PE)诊断、严重程度及妊娠结局评估中的价值。方法以分娩的150例PE孕妇(PE组)和100例健康孕妇(对照组)为研究对象,将PE组分为轻度PE(MPE组,n=81),重度PE(SPE组,n=69)。比较不同组别孕妇血清HDAC2、FOXA1水平,然后根据其HDAC2、FDXA1水平把PE孕妇分为HDAC2高表达组(n=72)和HDAC2低表达组(n=78),FOXA1高表达组(n=74)和FOXA1低表达组(n=76)。绘制受试者工作特征(ROC)曲线分析血清HDAC2、FOXA1对PE及不良妊娠结局的预测效能。Spearman相关性分析HDAC2、FOXA1与PE病情的关系。结果PE组血清HDAC2、FOXA1水平均明显低于对照组(P<0.05)。血清HDAC2、FOXA1及联合诊断PE的ROC曲线下面积(AUC)为0.863、0.843、0.912。MPE组血清HDAC2、FOXA1水平明显高于SPE组(P<0.05)。Spearman相关性分析显示,HDAC2、FOXA1均与PE严重程度呈负相关(r=-0.667、-0.712,均P=0.000)。对照组血清HDAC2、FOXA1水平明显高于PE组(P<0.05)。HDAC2低表达组早产儿、肝脏损害、新生儿窒息比例高于HDAC2高表达组(P<0.05)。FOXA1高表达组肝脏损害、早产儿、胸腹腔积液、新生儿窒息比例低于FOXA1低表达组(P<0.05)。血清HDAC2、FOXA1及联合预测PE孕妇不良妊娠结局的AUC为0.788、0.799、0.885。结论PE孕妇血清HDAC2、FOXA1水平与PE病情具有明显负相关性,低水平HDAC2、FOXA1的PE患者不良妊娠结局风险较高,临床可监测HDAC2、FOXA1来辅助诊断PE及预测妊娠结局。

外周血FOXO3a、VCAM-1水平评估2型糖尿病患者心血管病风险的价值

目的探讨外周血叉头盒O类转录因子-3a(Foxo3a)、血管细胞黏附分子-1(VCAM-1)评估2型糖尿病患者心血管病风险的价值。方法前瞻性选择2020年3月至2022年3月青岛市城阳区人民医院收治的2型糖尿病患者120例作为研究组,将同期体检的正常者50名作为对照组。采集所有入组者外周血,检测并对比两组FOXO3a、VCAM-1水平。将120例2型糖尿病患者按有无发生心血管疾病分为CVD组(n=45)及非CVD组(n=75),统计CVD组和非CVD组的一般资料、VCAM-1、FOXO3a水平等差异。采用单因素、多因素分析两组影响2型糖尿病患者发生心血管疾病的危险因素。采用受试者工作特征(ROC)评估外周血FOXO3a、VCAM-1水平预测2型糖尿病患者发生心血管疾病的价值。结果研究组患者VCAM-1水平(3.25±0.69)ng/mL,高于对照组[(1.76±0.59)ng/mL],FOXO3a水平为0.42±0.06,低于对照组(0.91±0.24),差异均有统计学意义(P<0.05)。CVD组体重指数≥24 kg/m2占比、有心血管疾病家族史占比、收缩压≥140 mmHg占比、VCAM-1、甘油三酯、总胆固醇水平为66.67%、57.78%、62.22%、(4.02±1.02)ng/mL、(2.67±0.52)mmol/L、(5.36±1.02)mmol/L,均高于非CVD组[33.33%、37.33%、41.33%、(3.06±0.98)ng/mL、(2.01±0.50)mmol/L、(4.12±1.01)mmol/L],FOXO3a水平0.33±0.09,低于非CVD组(0.54±0.12),差异均有统计学意义(P<0.05)。多因素Logisitic回归分析发现,体重指数≥24 kg/m2、有心血管疾病家族史、收缩压≥140 mmHg、VCAM-1≥3.817 ng/mL、FOXO3a≤0.440、甘油三酯≥2.338 mmol/L、总胆固醇≥4.483 mmol/L均是导致2型糖尿病患者发生心血管病的危险因素(P<0.05)。经过ROC曲线分析证实,VCAM-1、FOXO3a、甘油三酯、总胆固醇可用于2型糖尿病患者发生心血管病的预测,曲线下面积分别为0.735、0.936、0.893、0.833。结论2型糖尿病患者外周血VCAM-1、FOXO3a水平存在异常表达,可用于2型糖尿病患者发生心血管病的预测。

FOXO1基因多态性与2型糖尿病易感性的关联性分析

目的:探讨中国北方汉族人群叉头转录因子O1(FOXO1)基因多态性与2型糖尿病(T2DM)易感性的关系,阐明FOXO1基因与T2DM的关联性。方法:采用病例-对照研究设计,选取190例T2DM患者为病例组,193例健康人为对照组。应用基质辅助激光解吸附电离飞行时间质谱分析技术(MALDI-TOF-MS)检测FOXO1基因上rs2755213、rs2701880和rs10507486位点的多态性。结果:病例组和对照组rs2701880、rs2755213和rs10507486位点的等位基因和基因型频数分布差异均无统计学意义(P>0.05);3个位点组成的3种单倍型rs2755213-rs2701880、rs2701880-rs10507486、rs2755213-rs2701880-rs10507486在病例组和对照组中的分布差异均无统计学意义(P>0.05)。结论:未发现FOXO1基因rs2755213、rs2701880和rs10507486多态性与中国北方汉族人群T2DM的易感性存在关联。

2型糖尿病肾病患者血清miR-92b-5p和HMGB1水平变化及临床意义

目的观察miR-92b-5p及高迁移率族蛋白Box1(HMGB1)在2型糖尿病肾病患者血清中的水平变化,并探讨其临床意义。方法选取2型糖尿病患者203例,依据是否合并肾病将患者分为单纯糖尿病组(n=98)和糖尿病肾病组(n=105),并纳入80例同期体检健康者为对照组。采用酶联免疫吸附法检测各组血清HMGB1,采用实时荧光定量PCR法检测各组血清miR-92b-5p,采用多因素Logistic回归分析2型糖尿病肾病的危险因素,采用Pearson检验进行相关性分析,采用ROC曲线评估miR-92b-5p和HMGB1对2型糖尿病肾病的预测价值。结果单纯糖尿病组和糖尿病肾病组血清HMGB1水平高于对照组,且糖尿病肾病组高于单纯糖尿病组(P均<0.05)。单纯糖尿病组和糖尿病肾病组血清miR-92b-5p水平低于对照组,且糖尿病肾病组低于单纯糖尿病组(P均<0.05)。多因素Logistic回归分析显示,血清HMGB1水平升高与miR-92b-5p水平降低均是2型糖尿病进展为2型糖尿病肾病的独立危险因素(P均<0.05)。2型糖尿病患者血清miR-92b-5p水平与HMGB1和24 h尿蛋白(24 hpro)水平均呈负相关,与估算肾小球滤过率(eGFR)水平呈正相关(P均<0.05);HMGB1和24 hpro水平呈正相关,与eGFR水平呈负相关(P均<0.05)。血清HMGB1联合miR-92b-5p预测2型糖尿病肾病的ROC曲线下面积(AUC)为0.858(95%CI:0.800~0.908),高于miR-92b-5p(AUC=0.754,95%CI:0.688~0.821)、HMGB1(AUC=0.802,95%CI:0.737~0.860)单一检查(P均<0.05)。结论血清HMGB1水平升高和miR-92b-5p水平降低是2型糖尿病患者进展为糖尿病肾病的危险因素,早期联合检测两指标可作为临床预测及诊断2型糖尿病肾病的重要生化标志物。

下调FoxM1表达对人喉癌Hep-2细胞顺铂敏感性的影响

目的探讨下调Fox M1表达对人喉癌Hep-2细胞顺铂敏感性的影响及其可能机制。方法分别选用Fox M1 siRNA以及Fox M1特异性抑制剂(硫链丝菌肽)下调Hep-2细胞Fox M1表达。实时定量PCR、Western blot检测siRNA或硫链丝菌肽处理细胞后Fox M1 mRNA、蛋白表达量;MTT法检测下调Fox M1表达后Hep-2细胞的顺铂敏感性;流式细胞术检测下调Fox M1表达后顺铂诱导的凋亡率变化;实时定量PCR、Western blot检测顺铂作用于Fox M1下调组及对照组细胞,其Fox M1 mRNA、蛋白及凋亡相关蛋白Bcl-2、Bax表达变化。结果 siRNA及硫链丝菌肽均能下调Fox M1表达(P<0.05);两种方式均能降低顺铂作用下细胞存活率以及IC50[NC组顺铂IC50=(2.609±0.102)μg/m L,干扰组IC50=(0.771±0.058)μg/m L,P<0.05;对照组顺铂IC50=(2.142±0.198)μg/m L,抑制剂组IC50=(0.773±0.063)μg/m L,P<0.05],siRNA法处理后NC组、干扰组、NC+顺铂组、干扰+顺铂组凋亡率依次为(4.197±0.273)%、(12.553±0.183)%、(37.465±4.305)%、(82.373±7.214)%,干扰+顺铂组凋亡率显著升高(P<0.05);抑制剂处理后对照组、抑制剂组、顺铂组、抑制剂+顺铂组凋亡率依次为(2.343±0.194)%、(10.127±0.479)%、(35.075±1.995)%、(62.843±1.824)%,抑制剂+顺铂组凋亡率显著升高(P<0.05);下调Fox M1表达,凋亡抑制蛋白Bcl-2下调,促凋亡蛋白Bax表达上调(P<0.05)。结论下调Fox M1表达可提高Hep-2细胞对顺铂的敏感性,其机制可能与凋亡相关蛋白Bcl-2下调、Bax上调相关。