Oncogenic role of leptin and Notch interleukin-1 leptin crosstalk outcome in cancer

Obesity is a global pandemic characterized by high levels of body fat(adiposity) and derived-cytokines(i.e., leptin). Research shows that adiposity and leptin provide insight on the link between obesity and cancer progression. Leptin's main function is to regulate energy balance. However, obese individuals routinely develop leptin resistance, which is the consequence of the breakdown in the signaling mechanism controlling satiety resulting in the accumulation of leptin. Therefore, leptin levels are often chronically elevated in human obesity. Elevated leptin levels are related to higher incidence, increased progression and poor prognosis of several human cancers. In addition to adipose tissue, cancer cells can also secrete leptin and overexpress leptin receptors. Leptin is known to act as a mitogen, inflammatory and pro-angiogenic factor that induces cancer cell proliferation and tumor angiogenesis. Moreover, leptin signaling induces cancer stem cells, which are involved in cancer recurrence and drug resistance. A novel and complex signaling crosstalk between leptin, Notch and interleukin-1(IL-1) [Notch, IL-1 and leptin crosstalk outcome(NILCO)] seems to be an important driver of leptin-induced oncogenic actions. Leptin and NILCO signaling mediate the activation of cancer stem cells that can affect drug resistance. Thus, leptin and NILCO signaling are key links between obesity and cancer progression. This review presents updated data suggesting that adiposity affects cancer incidence, progression, and response to treatment. Here we show data supporting the oncogenic role of leptin in breast, endometrial, and pancreatic cancers.

Temporal Expression of Notch in Preterm Rat Lungs Exposed to Hyperoxia

Summary: To explore the mechanism of Notch in hyperoxia-induced preterm rat lung injury, 2-days-old preterm SD rats were randomized into control and hyperoxia group (FiO 2≥0.85). On day 1, 7, 14 and 21, 8 rat pups of each time point were used to assess histopathological changes of lung with HE staining and to evaluate the expression of Notch1 and Notch3 with immunohistochemistry. Notch1, Notch3, Aquaprin5 (AQP5) and surfactant protein C (SP-C) mRNA were measured by reverse transcription polymerase chain reaction (RT-PCR). The results showed that the lung injury in the hyperoxia group was characterized by retarded lung alveolization and differentiation of alveolar epithelial type Ⅱcells (AEC Ⅱ). Positive staining of Notch1 in hyperoxia group was weaker than controls at every time point (except for day 7), while positive staining of Notch3 was much stronger (P<0.05, P<0.01). Notch1, Notch3 mRNA level showed similar change as protein level. AQP5, SP-C mRNA decreased significantly as compared with that of the controls (P<0.01). We are led to conclude that hyperoxia results in abnormal expression of Notch, which is likely to contribute to the pathogenesis of lung injury through regulating proliferation and transdifferentiation of alveolar epithelial cells.

Elastic analysis of an elliptic notch in quasicrystals of point group 10 subjected to shear loading

Based on the stress potential and complex variable function method, this paper makes an elastic analysis of an elliptic notch subjected to uniform shear stress at infinity in quasicrystals with point group 10. With the aid of conformal transformation, an exact solution for the elliptic notch of the quasicrystals is obtained. The solution of the mode II Griffith crack as a special case is constructed. The stress intensity factor and energy release rate have been also obtained as a direct result of the crack solution.

THE EXPRESSION OF STRESS AND STRAIN AT THE TIP OF NOTCH IN REISSNER PLATE

In this paper, the eigenequation of notch in Reissner plate is derived by the eigenfunction method. Eigenvalues of different notches with different angles are calculated by Muller iteration method. The expression of stress and strain at the tip of notch in Reissner plate is obtained.

Notch1 mRNA和Dickkopf-1在评估非小细胞肺癌患者帕博利珠单抗治疗反应性中的价值

目的探讨Notch1 mRNA和Dickkopf-1在评估非小细胞肺癌(NSCLC)患者帕博利珠单抗治疗反应性中的价值。方法选取2020年7月—2022年9月邯郸市中心医院NSCLC患者169例(NSCLC组)、良性肺结节患者168例(对照组)。收集所有患者的临床资料,检测NSCLC组治疗前和帕博利珠单抗治疗3周、6周,对照组入组时的血清Notch1 mRNA相对表达量和Dickkopf-1水平。评估NSCLC患者帕博利珠单抗治疗6周后的治疗反应性[疾病进展(PD)、疾病控制(DC)]。采用受试者工作特征(ROC)曲线评价治疗前Notch1 mRNA、Dickkopf-1判断帕博利珠单抗治疗反应性的效能。采用Logistic回归模型分析帕博利珠单抗治疗反应性的影响因素。结果NSCLC组治疗前、治疗3周和治疗6周的血清Notch1 mRNA相对表达量和Dickkopf-1水平均高于对照组(P<0.001)。NSCLC组治疗前、治疗3周和治疗6周的血清Notch1 mRNA相对表达量和Dickkopf-1水平依次降低(P<0.001)。PD组与DC组之间年龄、美国东部肿瘤协作组(ECOG)评分、临床分期、淋巴转移、远隔转移、分化程度、程序性死亡受体配体1(PD-L1)表达和治疗前、治疗3周、治疗6周血清Notch1 mRNA相对表达量、Dickkopf-1水平差异均有统计学意义(P<0.05)。血清Notch1 mRNA和Dickkopf-1单项检测和联合检测评估帕博利珠单抗治疗反应性的ROC曲线下面积(AUC)分别为0.791、0.796、0.861。调整混杂因素后,治疗前高Notch1 mRNA组、高Dickkopf-1组PD风险分别是低Notch1 mRNA组、低Dickkopf-1组的3.517倍、3.326倍[比值比(OR)值分别为3.517、3.326,95%可信区间(CI)分别为2.159~5.728、2.211~5.003]。结论血清Notch1 mRNA表达和Dickkopf-1水平与帕博利珠单抗治疗反应性有关,可作为评估治疗反应性的有效指标。

miR-145通过Notch通路调节免疫功能及炎症反应介导创伤性脑损伤的神经保护

目的:探讨miR-145对创伤性脑损伤(TBI)后炎症反应和免疫调节的作用。方法:将雄性C57BL/6小鼠随机分为假手术组(Sham)、模型组(TBI)、TBI+NC agomir组、TBI+miR-145 agomir组。改良神经损伤严重程度评分(mNSS)用于评估创伤后神经功能;MWM测试评估TBI后小鼠的神经认知功能;流式细胞术检测各组小鼠脑组织中Tregs数量;ELISA检测各组小鼠海马中炎症细胞因子表达;免疫组化检测各组小鼠海马中活化的小胶质细胞/巨噬细胞Iba-1表达;RT-qPCR检测M1/M2小胶质细胞/巨噬细胞标志物基因iNOS、CD11b、CD206和Arg1表达;TUNEL染色和神经元细胞核免疫荧光标记(NeuN)的双重染色检测神经元凋亡。结果:与Sham组比较,TBI组小鼠海马组织中miR-145表达显著降低,神经功能损伤增加,脑组织中Tregs在CD4+T细胞群中百分比降低,海马组织中IL-1β、IL-6、TNF-α、IL-4、IL-10和TGF-β表达显著升高,活化的小胶质细胞/巨噬细胞Iba-1数量增多,iNOS、CD11b、CD206和Arg1表达水平显著升高,神经元凋亡升高,Notch1、p21和Hes1 mRNA和蛋白水平均显著升高(P<0.05);与TBI+NC agomir组比较,TBI+miR-145 agomir组小鼠海马中miR-145表达显著升高,神经功能损伤减轻,脑组织中Tregs在CD4+T细胞群中的百分比显著升高,海马中促炎因子IL-1β、IL-6、TNF-α表达显著降低,抗炎因子IL-4、IL-10和TGF-β表达显著升高,活化的小胶质细胞/巨噬细胞Iba-1数量显著减少,iNOS和CD11b表达降低,CD206和Arg1表达水平显著升高,神经元凋亡减少,Notch1、p21和Hes1 mRNA和蛋白水平均显著降低(均P<0.05)。结论:miR-145过表达通过提高Tregs水平,促进小胶质细胞M2极化,调节创伤后神经炎症反应及改善行为功能障碍,这一机制可能通过Notch信号通路介导。

miR-139-5p靶向Notch1抑制肺腺癌细胞增殖、迁移和侵袭

目的探究miR-139-5p在肺腺癌(lung adenocarcinoma,LUAD)进展中的作用和调控机制。方法实时定量PCR(qRT-PCR)和蛋白质印迹检测LUAD组织和细胞中miR-139-5p和Notch同源物1(Notch1)的表达。经细胞转染后,qRT-PCR和蛋白质印迹验证转染效率。CCK-8、细胞划痕和Transwell实验用于检测LUAD细胞的增殖、迁移和侵袭。双荧光素酶报告基因分析miR-139-5p与Notch1的靶向关系。蛋白质印迹检测PI3K/AKT/mTOR信号通路相关蛋白的表达。结果与正常组织和人正常支气管上皮细胞相比,miR-139-5p在LUAD组织和细胞中显著下调(P<0.01),Notch1显著上调(P<0.01)。与对照组相比,过表达miR-139-5p明显抑制LUAD细胞的增殖、迁移和侵袭,并下调p-PI3K、p-AKT和p-mTOR的表达(P<0.01)。Notch1被确定为miR-139-5p的直接靶标。过表达Notch1减弱了miR-139-5p过表达对LUAD细胞增殖、迁移和侵袭的抑制作用(P<0.05)。与对照组比较,miR-139-5p在体内显著抑制了异形移植瘤的生长(P<0.05)。结论miR-139-5p通过靶向Notch1并失活PI3K/Akt/mTOR途径抑制LUAD细胞的增殖、迁移和侵袭。

NOTCH3基因C260S位点突变导致CADASIL的临床和影像学特征分析

目的探讨NOTCH3基因第5外显子C260S位点突变导致的伴有皮层下梗死和白质脑病的常染色体显性遗传性脑动脉病(cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy,CADASIL)家系的临床和影像学特征。方法选取2021年12月首都医科大学附属北京同仁医院来自同一家庭的CADASIL患者,对所有患者进行NOTCH3基因测序,回顾性分析患者的临床表现和头颅影像学特征。复习既往文献报道的导致同一位置氨基酸改变的其他突变类型的临床及影像学特征。结果4名家庭成员中,包括先证者(46岁,女)及其两个姐姐(分别为48岁和50岁)和女儿(18岁)。先证者及其父亲、两个姐姐都有偏头痛病史,其中大姐有记忆力减退;先证者患有脑梗死及伴有视觉先兆的偏头痛;先证者女儿体健;先证者父亲因脑梗死去世。4名家庭成员均存在C260S位点的NOTCH3基因突变。既往文献无此位点突变的报道,先证者头颅MRI示右侧脑桥亚急性梗死,颞叶、脑室周围及脑干异常高信号改变,其大姐脑桥可见腔隙性梗死灶。结论NOTCH3基因第5外显子c.778T>A(p.C260S)的罕见突变导致的CADASIL发病时间早,早期会出现认知障碍。合并偏头痛的脑干梗死患者,需警惕CADASIL的可能。

Periostin、Notch1、维生素D与自身免疫性甲状腺炎淋巴细胞浸润程度、Treg/Th17的相关性研究

目的探讨骨外膜素(Periostin)、Notch跨膜受体-1(Notch1)m RNA、维生素D(VitD)与自身免疫性甲状腺炎(AIT)淋巴细胞浸润程度、调节性T细胞/辅助性T细胞17(Treg/Th17)的相关性。方法选取2021年7月至2023年12月郑州大学第一附属医院收治的92例AIT患者纳入AIT组,另选取同期50例无甲状腺疾病的健康人群纳入对照组。比较两组受检者的淋巴细胞浸润程度及抗体水平,采用Spearman、Pearson相关系数分析淋巴细胞浸润程度、Treg/Th17与甲状腺功能、抗体水平的相关性,比较两组受检者的Periostin、Notch1 m RNA、VitD及Treg/Th17,采用Pearson相关系数分析Periostin、Notch1 mRNA、VitD与淋巴细胞浸润程度及Treg/Th17的相关性。结果AIT组患者的CD3^(+)、CD3^(+)CD4^(+)、CD4^(+)CD25^(+)CD127^(-)、TgAb、TPOAb、TRAb水平及甲亢/亚临床甲亢、甲减/亚临床甲减患者占比明显高于对照组,差异均有统计学意义(P<0.05);Pearson相关系数分析结果显示,CD3^(+)(r=0.579、0.602、0.563)、CD3^(+)CD4^(+)(r=0.612、0.637、0.606)、CD~4+CD25^(+)CD127^(-)(r=0.655、0.643、0.687)与TgAb、TPOAb、TRAb呈正相关(P<0.05);AIT组患者的Periostin、Notch1 m RNA分别为(4.27±1.40)μg/L、1.73±0.56,明显高于对照组的(2.86±0.49)μg/L、1.02±0.14,VitD、Treg/Th17分别为(17.82±5.09)ng/mL、2.82±0.97,明显低于对照组的(22.30±3.76)ng/mL、12.36±2.03,差异均有统计学意义(P<0.05);Pearson相关系数分析结果显示,Periostin(r=0.792、0.811、0.737)、Notch1 mRNA(r=0.812、0.775、0.792)与CD3^(+)、CD3^(+)CD4^(+)、CD4^(+)CD25+CD127-呈正相关(P<0.05),VitD(r=-0.687、-0.753、-0.799)与之呈负相关(P<0.05),且Periostin(r=-0.823)、Notch1 m RNA(r=-0.772)与Treg/Th17呈负相关(P<0.05),VitD(r=0.745)与之呈正相关(P<0.05)。结论Periostin、Notch1 mRNA在AIT患者血清中表达上调,VitD表达下调,各指标与AIT淋巴细胞浸润程度及Treg/Th17均具有一定相关性,可为临床判断病情提供参考,并对后续临床治疗具有一定指导价值。

基于Notch与Wnt/β-catenin通路探讨黄芪甲苷对人成纤维细胞间质转化的作用

目的基于Notch与Wnt/β-catenin通路,探讨黄芪甲苷(astragalosideⅣ,AS-Ⅳ)对类风湿关节炎相关间质性肺病(rheumatoid arthritis associated interstitial lung disease,RA-ILD)体外模型的作用机制。方法用Luminex检测法对RA-ILD模型鼠支气管肺泡灌洗液中细胞因子进行检测,筛选出关键细胞因子;共培养人髓系白血病单核细胞(human myeloid leukemia mononuclear cells,THP-1)与正常人肺成纤维细胞(normal human lung fibroblast,NHLF)两种细胞系,加入关键细胞因子模拟RA-ILD体外模型,给予不同干预措施。通过RT-PCR、Western blot、COIP、EdU、ELISA以及双荧光素酶报告基因活性检测方法观察Snail、ZEB-1及E-Cadherin的启动子活性及mRNA表达、Notch通路中的关键蛋白表达、β-catenin的核转位水平、NHLF细胞增殖速率、ColⅠ、ColⅢ及Fibronectin的分泌水平以及β-catenin与NICD、β-catenin与LEF、NICD与CSL的相互作用。结果使用细胞因子处理NHLF细胞后,Snail、ZEB-1的启动子活性及mRNA表达明显增高,Notch通路中的关键蛋白表达水平显著上调,β-catenin的核转位明显增加,EdU阳性细胞明显增加;细胞上清中ColⅠ、ColⅢ及Fibronectin分泌水平显著上调,同时β-catenin与NICD、β-catenin与LEF、NICD与CSL的蛋白结合明显增加;使用Notch抑制剂DAPT和Jagged1 shRNA时结果与上述结论相反;AS-Ⅳ能够通过抑制Jagged1进而下调Notch及β-catenin信号通路的过度活化,抑制β-catenin与NICD、β-catenin与LEF、NICD与CSL的蛋白结合,能够剂量依赖性地抑制Snail及ZEB-1并上调E-Cadherin的启动子活性、mRNA和蛋白表达水平,抑制NHLF细胞的过度增殖及Col I、ColⅢ、Fibronectin的分泌。结论AS-Ⅳ可能通过抑制Notch和Wnt/β-catenin信号通路,抑制NHLF细胞增殖,减少ECM过度沉积,从而减缓RA-ILD肺间质纤维化进展。

雷公藤甲素通过miR-34b-5p/Notch1轴抑制骨肉瘤U2OS细胞增殖并诱导其铁死亡

目的:探究雷公藤甲素(TPL)通过miR-34b-5p调控Notch1表达对骨肉瘤U2OS细胞铁死亡影响的机制。方法:常规培养U2OS细胞,将其分为对照组、TPL(10μmol/L)组、TPL(10μmol/L)+Fer-1(铁死亡抑制剂,20μmol/L)组、miR-NC组、miR-34b-5p组、miR-34b-5p+Fer-1(20μmol/L)组、TPL(10μmol/L)+anti-miR-34b-5p组、anti-miR-34b-5p+Fer-1(20μmol/L)组。qPCR法、CCK-8法、铁离子检测试剂、DHE-荧光探针和WB法分别检测各组U2OS细胞中miR-34b-5p的表达、增殖能力、Fe2+水平、ROS水平以及铁死亡相关蛋白(GPX4、SLC7A11及Notch1蛋白)的表达,双萤光素酶报告基因实验验证miR-34b-5p与Notch1的靶向结合关系。结果:TPL可促进U2OS细胞中miR-34b-5p表达,Fer-1和anti-miR-34b-5p则抑制miR-34b-5p的表达(均P<0.05)。TPL明显抑制U2OS细胞的增殖、GPX4、SLC7A11、Notch1蛋白的表达、增加细胞中Fe2+和ROS的含量,Fer-1可逆转TPL对U2OS细胞的作用(均P<0.05)。过表达miR-34b-5p与TPL对U2OS细胞的作用相似(均P<0.05)。miR-34b-5p可靶向结合Notch1(均P<0.05)。miR-34b-5p抑制剂可明显抑制TPL对U2OS细胞的影响,Fer-1可增强miR-34b-5p抑制剂的作用(均P<0.05)。结论:TPL可抑制U2OS细胞的增殖能力并促进其铁死亡,其作用机制可能与miR-34a-5p靶向调节Notch1表达有关。

miR-34a-5p靶向Notch1对H/R诱导的人心肌细胞凋亡的影响及机制

目的探讨微小RNA(miR)-34a-5p/跨膜融合蛋白1(Notch1)信号轴介导内质网应激对缺氧/复氧(H/R)人心肌细胞的改善作用。方法人心肌细胞随机分为对照组(Control组)、缺氧复氧组(H/R组)、模拟物阴性对照组(mimic NC组)、模拟物组(mimic组)、抑制物阴性对照组(inhibitor NC组)、抑制物组(inhibitor组)。除Control组外,其余组细胞建立H/R损伤模型。定量反转录聚合酶链式反应(qRT-PCR)法检测miR-34a-5p和Notch1表达量,噻唑蓝(MTT)法检测细胞存活率,流式细胞仪检测细胞凋亡率,双荧光素酶基因报告法检测miR-34a-5p和Notch1的靶向关系,蛋白印迹法检测转录激活因子6(ATF6)、肌醇需求酶1(IRE1)、蛋白激酶样内质网激酶(PERK)以及葡萄糖调节蛋白78(GRP78)表达量。结果与Control组比较,H/R组miR-34a-5p表达量、细胞凋亡率以及ATF6、IRE1、PERK和GRP78蛋白表达量均升高,细胞存活率以及Notch1 mRNA和蛋白表达量均降低(P<0.05);与H/R组和mimic NC组比较,mimic组miR-34a-5p表达量、细胞凋亡率以及ATF6、IRE1、PERK和GRP78蛋白表达量均升高,细胞存活率以及Notch1 mRNA和蛋白表达量均降低(P<0.05);与H/R组和inhibitor NC组比较,inhibitor组miR-34a-5p表达量、细胞凋亡率以及ATF6、IRE1、PERK和GRP78蛋白表达量均降低,细胞存活率以及Notch1 mRNA和蛋白表达量均升高(P<0.05)。结论下调miR-34a-5p可抑制H/R人心肌细胞凋亡,其可能是通过靶向激活Notch1介导内质网应激发挥作用。

五味子甲素调节miR-429/Notch1信号轴对冠心病大鼠心肌组织损伤的影响

目的:探究五味子甲素(Sch A)调节miR-429/Notch1信号轴对冠心病大鼠心肌组织损伤的影响。方法:利用盐酸异丙肾上腺素(ISO)诱导建立冠心病大鼠模型,随机分为对照(Control)组、模型(ISO)组、低剂量Sch A组(L-Sch A组)、中剂量Sch A组(M-Sch A组)、高剂量Sch A组(H-Sch A组)、普萘洛尔(Pro)组、miR-429 mimics阴性对照+H-Sch A组、miR-429 mimics+H-Sch A组。于ISO诱导30 min后,观察心功能指标[心率、平均动脉压(MAP)、左心室收缩压(LVSP)、左心室舒张末压(LVEDP)];酶联免疫吸附试验(ELISA)检测心肌损伤血清标志物[肌酸激酶同工酶MB(CK-MB)、心肌肌钙蛋白I(cTnI)、B型钠尿肽(BNP)];苏木精-伊红(HE)染色观察心肌组织病理损伤;实时荧光定量聚合酶链式反应(RT-qPCR)检测心肌组织miR-429 mRNA;蛋白免疫印迹法(Western Blot)检测心肌组织Notch1信号相关因子[Notch受体胞内结构域(NICD)、Hes1]蛋白。结果:与Control组比较,ISO组心率、MAP、LVSP降低(P<0.05),LVEDP、CK-MB、cTnI、BNP升高(P<0.05),同时可见明显病理学损伤,心肌细胞结构异常、肌原纤维排列紊乱、炎性细胞浸润;而经L-Sch A组、M-Sch A组、H-Sch A组及Pro组心率、MAP、LVSP升高(P<0.05),LVEDP、CK-MB、cTnI、BNP降低(P<0.05),同时病理学损伤明显减轻,且H-Sch A效果更佳,与Pro效果相当。与Control组比较,ISO组miR-429 mRNA升高(P<0.05),NICD/甘油醛-3-磷酸脱氢酶(GAPDH)、Hes1/GAPDH蛋白比值降低(P<0.05);而经H-Sch A预处理,miR-429 mRNA降低,NICD/GAPDH、Hes1/GAPDH蛋白比值升高(P<0.05);而使miR-429过表达后,NICD/GAPDH、Hes1/GAPDH蛋白比值降低(P<0.05);同时LVEDP、CK-MB、cTnI、BNP升高,心率、MAP、LVSP降低(P<0.05),以及心肌组织病理损伤程度有所加重。结论:Sch A可减轻冠心病大鼠心肌组织损伤,可能通过下调miR-429表达,进而激活Notch1信号通路而实现。

调脾安肠方对裸鼠结肠癌肝转移灶miR-34a-5p、Notch1/NF-κB表达的影响

目的探讨调脾安肠方对裸鼠结肠癌肝转移(colorectal cancer liver metastasis,CLM)灶组织中MicroRNA-34a-5p(miR-34a-5p)及其靶基因Notch1/核转录因子-κB(nuclear factor-kappa B,NF-κB)通路表达的影响。方法选取30只雄性Balb/c裸鼠随机分为空白组、模型组、中药组,每组10只。脾脏原位注射HCT-116人结肠癌细胞(细胞数约为1×107个/mL)法构建CLM模型。中药组在CLM造模完成2周后开始调脾安肠方(2 g/mL,每日0.2 mL)灌胃2周,空白组和模型组同期予等体积0.9%生理盐水灌胃。苏木素—伊红染色观察肝转移灶病理组织形态;蛋白免疫印迹法测定肝转移灶Notch1、NF-κB p65蛋白水平;实时荧光定量PCR测定肝转移灶miR-34a-5p及Notch1、NF-κB p65 mRNA水平。结果与模型组比较,中药组裸鼠镜下病理转移灶面积明显减小,核分裂像减少,提示调脾安肠方抑制了结直肠癌细胞在肝脏的定植和迁移。与空白组比较,模型组肝转移灶中miR-34a-5p含量显著降低(P<0.01),Notch1、NF-κB p65蛋白表达升高(均P<0.05),Notch1、NF-κB p65的mRNA表达显著升高(均P<0.01)。与模型组比较,中药组Notch1蛋白及mRNA表达降低(P<0.05),NF-κB p65的mRNA表达显著降低(P<0.01)。miR-34a-5p与Notch1 mRNA在结肠癌肝转移灶组织中表达水平呈负相关(r^(2)=0.8845,P<0.05),Notch1与NF-κB p65在结肠癌肝转移灶组织中mRNA表达水平呈显著正相关(r^(2)=0.9291,P<0.01)。结论中药复方调脾安肠方能抑制裸鼠结肠癌肝转移灶形成,其机制可能与通过抑癌基因miR-34a-5p靶向负性调节Notch1/NF-κB通路表达有关。

A Wnt10a-Notch signaling axis controls Hertwig’s epithelial root sheath cell behaviors during root furcation patterning

Human with bi-allelic WNT10A mutations and epithelial Wnt10a knockout mice present enlarged pulp chamber and apical displacement of the root furcation of multi-rooted teeth,known as taurodontism;thus,indicating the critical role of Wnt10a in tooth root morphogenesis.However,the endogenous mechanism by which epithelial Wnt10a regulates Hertwig’s epithelial root sheath(HERS)cellular behaviors and contributes to root furcation patterning remains unclear.In this study,we found that HERS in the presumptive root furcating region failed to elongate at an appropriate horizontal level in K14-Cre;Wnt10a^(fl/fl)mice from post-natal day 0.5(PN0.5)to PN4.5.EdU assays and immunofluorescent staining of cyclin D1 revealed significantly decreased proliferation activity of inner enamel epithelial(IEE)cells of HERS in K14-Cre;Wnt10a^(fl/fl)mice at PN2.5 and PN3.5.Immunofluorescent staining of E-Cadherin and acetyl-α-Tubulin demonstrated that the IEE cells of HERS tended to divide perpendicularly to the horizontal plane,which impaired the horizontal extension of HERS in the presumptive root furcating region of K14-Cre;Wnt10a^(fl/fl)mice.RNA-seq and immunofluorescence showed that the expressions of Jag1 and Notch2 were downregulated in IEE cells of HERS in K14-Cre;Wnt10a^(fl/fl)mice.Furthermore,after activation of Notch signaling in K14-Cre;Wnt10a^(fl/fl)molars by Notch2 adenovirus and kidney capsule grafts,the root furcation defect was partially rescued.Taken together,our study demonstrates that an epithelial Wnt10a-Notch signaling axis is crucial for modulating HERS cell proper proliferation and horizontal-oriented division during tooth root furcation morphogenesis.

Prox1 Suppresses Proliferation and Drug Resistance of Retinoblastoma Cells via Targeting Notch1

Objective Retinoblastoma(RB)is a prevalent type of eye cancer in youngsters.Prospero homeobox 1(Prox1)is a homeobox transcriptional repressor and downstream target of the proneural gene that is relevant in lymphatic,hepatocyte,pancreatic,heart,lens,retinal,and cancer cells.The goal of this study was to investigate the role of Prox1 in RB cell proliferation and drug resistance,as well as to explore the underlying Notch1 mechanism.Methods Human RB cell lines(SO-RB50 and Y79)and a primary human retinal microvascular endothelial cell line(ACBRI-181)were used in this study.The expression of Prox1 and Notch1 mRNA and protein in RB cells was detected using quantitative real time-polymerase chain reaction(RT-qPCR)and Western blotting.Cell proliferation was assessed after Prox1 overexpression using the Cell Counting Kit-8 and the MTS assay.Drug-resistant cell lines(SO-RB50/vincristine)were generated and treated with Prox1 to investigate the role of Prox1 in drug resistance.We employed pcDNA-Notch1 to overexpress Notch1 to confirm the role of Notch1 in the protective function of Prox1.Finally,a xenograft model was constructed to assess the effect of Prox1 on RB in vivo.Results Prox1 was significantly downregulated in RB cells.Overexpression of Prox1 effectively decreased RB cell growth while increasing the sensitivity of drug-resistant cells to vincristine.Notch1 was involved in Prox1’s regulatory effects.Notch1 was identified as a target gene of Prox1,which was found to be upregulated in RB cells and repressed by increased Prox1 expression.When pcDNA-Notch1 was transfected,the effect of Prox1 overexpression on RB was removed.Furthermore,by downregulating Notch1,Prox1 overexpression slowed tumor development and increased vincristine sensitivity in vivo.Conclusion These data show that Prox1 decreased RB cell proliferation and drug resistance by targeting Notch1,implying that Prox1 could be a potential therapeutic target for RB.

Direct Pointwise Comparison of FE Predictions to StereoDIC Measurements:Developments and Validation Using Double Edge-Notched Tensile Specimen

To compare finite element analysis(FEA)predictions and stereovision digital image correlation(StereoDIC)strain measurements at the same spatial positions throughout a region of interest,a field comparison procedure is developed.The procedure includes(a)conversion of the finite element data into a triangular mesh,(b)selection of a common coordinate system,(c)determination of the rigid body transformation to place both measurements and FEA data in the same system and(d)interpolation of the FEA nodal information to the same spatial locations as the StereoDIC measurements using barycentric coordinates.For an aluminum Al-6061 double edge notched tensile specimen,FEA results are obtained using both the von Mises isotropic yield criterion and Hill’s quadratic anisotropic yield criterion,with the unknown Hill model parameters determined using full-field specimen strain measurements for the nominally plane stress specimen.Using Hill’s quadratic anisotropic yield criterion,the point-by-point comparison of experimentally based full-field strains and stresses to finite element predictions are shown to be in excellent agreement,confirming the effectiveness of the field comparison process.

先天性心脏病患儿母体孕期血清miR-22、Notch1表达及临床意义

目的:分析先天性心脏病患儿母体孕期血清miR-22、Notch1表达及临床意义。方法:选取苏州吴江地区2019年1月—2020年12月出生的先天性心脏病患儿的孕妇58例为观察组,选取同期住院分娩正常儿童的孕妇308名为对照组。对两组孕妇一般资料及环境暴露因素进行问卷调查,采用单因素及二分类Logistic回归分析潜在暴露因素与先天性心脏病之间的潜在关系;使用逆转录-聚合酶链式反应(RT-PCR)及Pearson分析miR-22及Notch1表达及潜在关系。结果:观察组外周血清miR-22低于对照组,Notch1高于对照组,差异有统计学意义(P<0.05)。Pearson分析显示,miR-22与Notch1呈负相关(P<0.05)。两组孕妇孕期体重增加>15 kg、亚临床甲状腺功能减退、孕早中期呼吸道感染、妊娠期合并症、喜吃烧烤比较,差异均有统计学意义(P<0.05)。Logistic分析结果显示,孕期体重增加>15 kg、亚临床甲状腺功能减退、孕早中期呼吸道感染、妊娠期合并症、喜吃烧烤、miR-22水平降低及Notch1升高为新生儿先天性心脏病发生的独立危险因素。结论:miR-22及Notch1异常表达与先天性心脏病发生密切相关,具有较好的研究价值;孕期多种暴露因素与先天性心脏病的发生发展相关,临床工作应积极指导孕期健康,以降低先天性心脏病发生率。

变应性鼻炎患者血清miR-149-5p、Notch1 mRNA表达与Treg/Th17平衡的关系

目的探讨变应性鼻炎(AR)患者血清miR-149-5p、Notch1 mRNA表达与调节性T细胞(Treg)/辅助性T细胞(Th)17平衡的关系。方法选择98例AR患者(AR组)和30例健康体检志愿者(对照组),根据疾病严重程度将AR患者分为轻度组(51例)、中重度组(47例)。采用实时荧光定量聚合酶链反应检测血清miR-149-5p、Notch1 mRNA表达,采用流式法检测外周血中Treg、Th17及其细胞因子。Pearson相关性分析血清miR-149-5p、Notch1 mRNA表达与外周血中Treg、Th17及其细胞因子以及miR-149-5p与Notch1 mRNA表达的相关性。结果AR组血清miR-149-5p表达,白细胞介素(IL)-10、转化生长因子-β(1 TGF-β1)水平以及外周血Treg细胞、Treg/Th17值低于对照组(P均<0.05),血清Notch1 mRNA表达,IL-6、IL-17、IL-22、IL-23水平以及外周血Th17细胞高于对照组(P均<0.05)。中重度组血清miR-149-5p表达,IL-10、TGF-β1水平以及外周血Treg细胞、Treg/Th17值低于轻度组(P均<0.05),血清Notch1 mRNA表达,IL-6、IL-17、IL-22、IL-23水平以及外周血Th17细胞高于轻度组(P均<0.05)。AR组血清miR-149-5p表达与外周血Treg细胞、Treg/Th17值、血清IL-10、TGF-β1水平呈正相关(r分别为0.367、0.539、0.265、0.301,P均<0.05),与血清IL-6、IL-17、IL-22、IL-23水平以及外周血Th17细胞呈负相关(r分别为-0.289、-0.213、-0.336、-0.243、-0.221,P均<0.05)。Notch1 mRNA表达与外周血Treg细胞、Treg/Th17值、血清IL-10、TGF-β1水平呈负相关(r分别为-0.346、-0.508、-0.281、-0.293,P均<0.05),与血清IL-6、IL-17、IL-22、IL-23水平以及外周血Th17细胞呈正相关(r分别为0.326、0.241、0.302、0.285、0.237,P均<0.05)。血清miR-149-5p与Notch1 mRNA表达呈负相关(r=-0.550,P<0.05)。结论AR患者血清miR-149-5p表达下调,Notch1 mRNA表达上调,且与AR患者Treg/Th17失衡有关。

四味土木香散通过调节miR-34a-5p/Notch1信号通路改善机械牵张诱导的H9c2心肌细胞肥大

目的探讨四味土木香散(siwei tumuxiang powder,STP)对机械牵张诱导的大鼠H9c2心肌细胞肥大模型的保护作用及作用机制。方法制备STP含药血清及细胞肥大模型,并筛选STP含药血清最适剂量与作用时间;检测H9c2心肌细胞中心房钠尿肽(atrial natriuretic peptide,ANP)、脑钠肽(brain natriuretic peptide,BNP)、miR-34a-5p及Notch1等因子的表达,验证STP是否通过调控miR-34a-5p/Notch1通路对H9c2心肌细胞起保护作用。结果STP含药血清干预模型细胞最适剂量为低剂量(0.972g/kg),作用时间为24h。STP降低了模型细胞中ANP、BNP、miR-34a-5p的表达(P<0.05)。转染miR-34a-5p mimic逆转了STP降低ANP、BNP表达的作用(P<0.05)。用siRNA法敲低Notch1逆转了STP降低ANP、BNP的表达(P<0.05)。Notch1为miR-34a-5p直接靶基因,miR-34a-5p降低了Notch1的表达(P<0.05)。STP干预后,Notch1、NICD1、Hes1蛋白表达降低(P<0.05);转染miR-34a-5p mimic逆转了STP降低Notch1、NICD1、Hes1蛋白表达的作用(P<0.05)。结论STP可能是通过影响miR-34a-5p表达负调控Notch1信号通路对机械牵张诱导的大鼠H9c2心肌细胞肥大发挥保护作用。