非小细胞肺癌中Notch1、HIF-1、VEGF蛋白及Notch1 mRNA的表达及意义

目的探讨Notch1、HIF-1、VEGF蛋白及Notch1 mRNA在非小细胞肺癌(non-small cell lung cancer,NSCLC)组织的表达及其临床病理学意义。方法采用免疫组化SP法和原位杂交法分别检测65例NSCLC组织、15例正常支气管上皮组织中Notch1、HIF-1、VEGF蛋白及Notch1 mRNA的表达。结果Notch1、HIF-1、VEGF蛋白及Notch1 mRNA在非小细胞肺癌中阳性表达率分别为81.5%(53/65)、96.9%(63/65)、93.8%(61/65)、73.8%(48/65),均明显高于正常支气管上皮组织阳性表达率(P<0.05);NSCLC中Notch1、HIF-1、VEGF蛋白及Notch1 mRNA的表达均与临床分期、淋巴结转移有关(P<0.05);Notch1、HIF-1与VEGF蛋白间均正相关;Notch1蛋白与Notch1 mRNA的表达呈正相关。结论Notch1、HIF-1、VEGF蛋白及Notch1 mRNA在NSCLC中均表达上调,提示在肺癌的发生、发展中可能起重要作用;检测NSCLC组织Notch1蛋白及mRNA可作为判断肿瘤侵袭与转移的重要指标。

Notch1和HIF-1α蛋白在胃癌中的表达及临床意义

目的探讨Notch1和HIF-1α蛋白在胃癌中的表达与临床病理特征及预后关系。方法应用组织芯片技术、免疫组化染色,检测Notch1和HIF-1α在135例胃癌及27例正常胃黏膜中的表达。对正常胃黏膜与胃癌Notch1和HIF-1α蛋白阳性表达率的差异以及与临床病理特征的关系,Notch1和HIF-1α蛋白阳性表达的相关性,Notch1和HIF-1α蛋白阳性表达与生存期的关系运用统计学方法进行显著性检验。结果Notch1和HIF-1α蛋白在胃癌组织中阳性表达率高于正常胃黏膜(P<0.05);TNM分期中的Ⅲ-Ⅳ期组和伴有淋巴结转移组的Notch1和HIF-1α阳性表达率高于相应的Ⅰ-Ⅱ期组和无淋巴结转移组(P<0.05);Notch1与HIF-1α间存在正相关关系(P<0.01),并与生存期密切相关,阳性表达组较之阴性表达组平均生存期缩短(P<0.05)。结论Notch1和HIF-1α蛋白阳性表达与胃癌侵袭、转移密切相关,对于判断胃癌的预后可能具有重要意义。

Notch信号通路中HIF-1α及Notch-1在组织瓣缺血再灌注损伤后的表达变化

目的 :研究缺氧诱导的Notch信号通路相关因子在组织瓣缺血再灌注损伤后的表达变化,探讨其在组织瓣缺血再灌注损伤后损伤修复中可能的作用。方法:将24只Wistar大鼠随机分为空白对照组(Blank组)、假手术组(Sham组)和实验组(即缺血再灌注损伤组,I/R组),其中空白对照组4只,Sham组4只,I/R组16只。Sham组及I/R组制备大鼠左下腹部4 cm×8 cm皮瓣,I/R组以血管夹夹闭大鼠腹壁浅动、静脉,形成皮瓣缺血状态。Sham组于术后7 d,I/R组分别于去除血管夹(缺血6 h后)即刻及缺血后1、3、7 d过量麻醉处死。利用RT-PCR、Western免疫印迹分析HIF-1α、Notch-1的m RNA扩增水平和蛋白水平。H-E染色观察Sham组及I/R组术后第7天的组织变化。利用SPSS 19.0软件包对数据进行统计学分析。结果:H-E染色结果显示,术后第7天,I/R组较对照组及假手术组可见明显结构紊乱。RT-PCR结果显示,I/R组的HIF-1α、Notch-1 m RNA扩增水平在缺血6 h后即刻及缺血后1、3、7 d均显著高于Sham组及Blank组(P<0.05);Western免疫印迹结果显示,在I/R组,HIF-1α、Notch-1蛋白水平在缺血6 h后即刻及缺血后1、3、7 d均显著高于Sham组及Blank组(P<0.05)。结论:组织瓣在缺血再灌注后,组织出现明显损伤。Notch信号通路在损伤及修复再生过程中被激活,可能与组织缺氧引起的HIF-1α表达升高有关。

肾癌组织Notch 1、HIF-1α的表达及其与肿瘤临床分期的相关性

目的:探讨肾癌组织Notch1、HIF-1α的表达及意义。方法:应用组织芯片、免疫组化染色法比较肾癌(n=60)及正常肾组织(n=30)中Notch1、HIF-1α蛋白的表达差异,分析二者表达与肿瘤临床分期的相关性。结果:肾癌组织Notch1、HIF-1α阳性表达率高于正常肾组织(40.0%vs13.3%,58.3%vs0%,P<0.05);TNM分期Ⅲ～Ⅳ期肾癌组织Notch1、HIF-1α阳性表达率高于Ⅰ～Ⅱ期(27.5%vs65.0%,45.0%vs85.0%,P<0.05)。结论:肾癌组织中Notch1、HIF-1α表达均高于正常肾组织,且与肿瘤临床分期有关,二者对判断临床预后可能具有一定价值。

HIF-1α、Notch1及上皮间质转化在乳腺癌中的表达及意义

目的探讨缺氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)、Notch1与上皮间质转化(epithelial-mesenchymal transition,EMT)免疫表型在乳腺浸润性导管癌中的表达及临床意义。方法采用免疫组化法,检测乳腺浸润性导管癌(60例)和癌旁正常乳腺组织(20例)中HIF-1α、Notch1及EMT免疫表型E-钙粘蛋白(E-cadherin)、波形蛋白(Vimentin)的表达。结果 HIF-1α、Notch1、E-cadherin、Vimentin在乳腺癌组织中的表达阳性率分别为78.3%、81.7%、75%、33.3%,与癌旁正常乳腺组织中表达(阳性率分别为10%、20%、100%、0%)差异具有统计学意义(P<0.05);HIF-1α、Notch1、E-cadherin、Vimentin的表达与浸润性导管癌组织学分级、腋窝淋巴结转移及临床分期密切相关(P<0.05)。HIF-1α的表达与Notch1、Vimentin的表达呈正相关关系(P<0.05),而与E-cadherin的表达呈负相关关系(r=-0.545,P<0.05)。结论 HIF-1α通过上调Notch1、Vimentin的表达,下调E-cadherin的表达,促进乳腺浸润性导管癌EMT的发生,并促进癌组织的转移。联合检测HIF-1α、Notch1、E-cadherin、Vimentin的表达可成为判断乳腺癌生物学行为及转移情况的重要参考指标。

Protective Effect of Catalpol on Myocardium in Rats with Isoprenaline-Induced Myocardial Infarcts via Angiogenesis through Endothelial Progenitor Cells and Notch1 Signaling Pathway

Protective effect of catalpol on myocardium was studied in relation to endothelial progenitor cells, Notch1 signaling pathway and angiogenesis in rats with isoprenaline (INN)-induced acute myocardial infarcts. To analyze the pathological status and impact of catalpol on the rats, 3 weeks after intragastric gavage, the animals were verified for myocardial infarcts with electrocardiogram and measured for enzyme activity of lactate dehydrogenase (LDH), malondialdehyde (MDA), creatine kinase (CK) and superoxide dismutase (SOD) in myocardium, and further analyzed using HE and TTC staining, as well as visual examination of infarct area. Flow cytometry study of endothelial progenitor cells (EPCs) indicated that the EPCs were mobilized during infarction. The roles of Notch1 signaling pathway in angiogenesis of the infracted animals were studied using immunohistochemistry analysis of RBPjκ and Western blot analysis of Notch1 and Jagged1. Our results obtained from the rats treated with catalpol, positive drug and control showed that catalpol could protect rats from infarction probably by mobilization of EPCs and activation of Notch1 signaling pathway.

电针预处理大鼠百会穴对脑缺血保护作用及HIF-1α相关机制的研究

目的:旨在证实电针预处理百会穴对脑缺血损伤的保护作用及HIF-1α的相关机制。方法:将雄性SD大鼠80只随机分为5组(n=16):假手术组(Sham),对照组(CON),电针预处理组(EA),HIF-1α抑制剂组(2ME2)和电针预处理+HIF-1α抑制剂组(EA+2ME2)。CON组、2ME2组、EA组及EA+2ME2组动物均建立大脑中动脉阻闭模型(MCAO),Sham组动物仅接受同前手术操作;EA组及EA+2ME2组大鼠接受连续5 d电针刺激后24 h建立MCAO模型;2ME2组及EA+2ME2组动物分别于MCAO模型制备前30 min腹腔注射16 mg/kg的2ME2。缺血再灌注后24 h,各组随机抽取8只动物处死,行TUNEL染色检测神经元凋亡,Western Blot检测缺血半暗带中Bcl2/Bax的表达;缺血再灌注后72 h,各组另8只动物接受神经功能学评分后行磁共振成像(MRI)检查,随后处死行TTC染色检测脑梗死容积。结果:EA组脑梗死容积百分比显著低于CON组(P<0.05);与CON组比较,EA组神经功能评分显著改善(P<0.05),而2ME2可以显著降低EA+2ME2组神经功能评分(P<0.05)。与CON组比较,EA组神经功能评分显著改善(P<0.05),而2ME2可以显著降低EA+2ME2组神经功能评分(P<0.05)。而CON组及EA+2ME2组中TUNEL阳性细胞显著多于EA组(P<0.05)。与EA组比较,CON组及EA+2ME2组中Bcl-2蛋白表达显著降低(P<0.05),而CON组中Bax蛋白表达显著增加(P<0.05)。结论:电针预处理百会穴对脑缺血再灌注损伤具有明显的保护作用,其机制可能是HIF-1α抑制缺血后神经凋亡、上调凋亡抑制蛋白Bcl-2表达、下调促凋亡蛋白Bax表达,达到脑缺血保护的作用。

针刺经筋点对膝关节骨性关节炎Notch信号通路调控机制及Notch1、Notch2、Jagged1、Jagged2蛋白表达的实验研究

目的 通过针刺经筋点对家兔膝关节骨性关节炎(knee osteoarthritis, KOA)的干预治疗,观察家兔治疗前后血清中缺氧诱导因子2α(HIF-2α)、膝关节软骨细胞中Notch1、Notch2、以及Jagge1、Jagged2蛋白的表达情况,以探讨在经筋理论指导下的针刺方法对膝关节炎家兔的Notch信号通路的调控机制。方法 选取30只清洁级日本大耳白兔,雌雄各半,随机数字表法抽取8只正常饲养做为空白组,余下兔均按左后肢伸直位石膏固定制动法(Vidman法)制造兔膝关节炎模型。制动满6周时,拆除固定,随机选取空白组和造模家兔各2只,应用量角器检测膝关节关节活动度,并处死取关节软骨作病理观察。将造模后的家兔随机分为模型组、经筋组、西药组,6只/组,并分别于干预4周后处死取样。用量角器和改良Lequesne MG评分标准分别测量和观察治疗前后兔膝关节的关节活动度(ROM)和行为学指标,取关节软骨标本并观测其形态学改变,ELISA法检测血清中HIF-2α含量,并采用Western Blot法检测Notch1、Notch2、以及Jagged1、Jagged2蛋白表达量。结果 治疗后,经筋组和西药组的标本中HIF-2α、Notch1、Notch2、以及Jagged1、Jagged2蛋白的阳性表达率均低于模型组,但略高于空白组(P<0.01)。结论 经筋理论指导下,针刺对膝骨关节炎家兔有良好的疗效,其作用机制可能是降低HIF-2α、受体Notch1、Notch2、以及配体Jagged1、Jagged2蛋白的表达从而对Notch信号通路产生抑制作用,以减轻家兔膝骨关节炎的发展,为临床提供理论支持。

Negative effects of Notch1 on the differentiation of muscle-derived stem cells into neuronal-like cells

We cultured rat muscle-derived stem cells in medium containing nerve growth factor and basic fi-broblast growth factor to induce neuronal-like cell differentiation.Immunocytochemical staining and reverse transcription-PCR showed that the differentiated muscle-derived stem cells exhibited processes similar to those of neuronal-like cells and neuron-specific enolase expression,but Notch1 mRNA and protein expression was decreased.Down-regulation of Notch1 expression may facilitate neuronal-like cell differentiation from muscle-derived stem cells.

Buyang Huanwu decoction up-regulates Notch1 gene expression in injured spinal cord

Expression of genes in the Notch signaling pathway is altered in the injured spinal cord, which indicates that Notch participates in repair after spinal cord injury. Buyang Huanwu decoction, a traditional Chinese herbal preparation, can promote the growth of nerve cells and nerve fibers; however, it is unclear whether Buyang Huanwu decoction affects the Notch signaling pathway in injured spinal cord. In this study, a rat model was established by injuring the T10 spinal cord. At 2 days after injury, rats were intragastrically administered 2 m L of 0.8 g/m L Buyang Huanwu decoction daily until sacrifice. Real-time reverse transcription polymerase chain reaction analysis demonstrated that at 7, 14 and 28 days after injury, the expression of Notch1 was increased in the Buyang Huanwu decoction group compared with controls. These findings confirm that Buyang Huanwu decoction can promote the expression of Notch1 in rats with incomplete spinal cord injury, and may indicate a mechanism to promote the repair of spinal cord injury.

Oncogenic potential of IDH1R132C mutant incholangiocarcinoma development in mice

AIM: To investigate whether IDH1R132 C mutant in combination with loss of p53 and activated Notch signaling promotes intrahepatic cholangiocarcinoma(ICC) development.METHODS: We applied hydrodynamic injection and sleeping beauty mediated somatic integration to induce loss of p53(via sh P53), activation of Notch [via intracellular domain of Notch1(NICD)] and/or overexpression of IDH1R132 C mutant together with the sleeping beauty transposase into the mouse liver. Specifically, we co-expressed sh P53 and NICD(sh P53/NICD, n = 4), sh P53 and IDH1R132C(sh P53/IDH1R132 C, n = 3), NICD and IDH1R132C(NICD/IDH1R132 C, n = 4), as well as NICD, sh P53 and IDH1R132C(NICD/sh P53/IDH1R132 C, n = 9) in mice. Mice were monitored for liver tumor development and euthanized at various time points. Liver histology was analyzed by hematoxylin and eosin staining. Molecular features of NICD/sh P53/IDH1R132 C ICC tumor cells were characterized by Myc tag, Flag tag, Ki-67, p-Erk and p-AKT immunohistochemical staining. Desmoplastic reaction in tumor tissues was studied by Picro-Sirius red staining.RESULTS: We found that co-expression of sh P53/NICD, sh P53/IDH1R132 C or NICD/IDH1R132 C did not lead to liver tumor formation. In striking contrast, coexpression of NICD/sh P53/IDH1R132 C resulted in ICC development in mice(P < 0.01). The tumors could be identified as early as 12 wk post hydrodynamic injection. Tumors rapidly progressed, and by 18 wk post hydrodynamic injection, multiple cystic lesions could be identified on the liver surface. NICD/sh P53/IDH1R132 C liver tumors shared multiple histological features of human ICCs, including hyperplasia of irregular glands. Importantly, all tumor cells were positive for the biliary epithelial cell marker cytokeratin 19. Extensive collagen fibers could be visualized in tumor tissues using Sirus red staining, duplicating the desmoplastic reaction observed in human ICC. Tumors were highly proliferative and expressed ectopically injected genes. Together these studies supported that NICD/sh P53/IDH1R132 C liver tumors were indeed ICCs. Finally, no p-AKT or p-ERK positive staining was observed, suggesting that NICD/sh P53/IDH1R132 C driven ICC development was independent of AKT/m TOR and Ras/MAPK signaling cascades. CONCLUSION: We have generated a simple, nongermline murine ICC model with activated Notch, loss of p53 and IDH1R132 C mutant. The study supported the oncogenic potential of IDH1R132 C.

Zinc oxide nanoparticles inhibit osteosarcoma metastasis by downregulatingβ-catenin via HIF-1α/BNIP3/LC3B-mediated mitophagy pathway

Osteosarcoma(OS)therapy faces many challenges,especially the poor survival rate once metastasis occurs.Therefore,it is crucial to explore new OS treatment strategies that can efficiently inhibit OS metastasis.Bioactive nanoparticles such as zinc oxide nanoparticles(ZnO NPs)can efficiently inhibit OS growth,however,the effect and mechanisms of them on tumor metastasis are still not clear.In this study,we firstly prepared well-dispersed ZnO NPs and proved that ZnO NPs can inhibit OS metastasis-related malignant behaviors including migration,invasion,and epithelial-mesenchymal transition(EMT).RNA-Seqs found that differentially expressed genes(DEGs)in ZnO NP-treated OS cells were enriched in wingless/integrated(Wnt)and hypoxia-inducible factor-1(HIF-1)signaling pathway.We further proved that Zn^(2+)released from ZnO NPs induced downregulation ofβ-catenin expression via HIF-1α/BNIP3/LC3B-mediated mitophagy pathway.ZnO NPs combined with ICG-001,aβ-catenin inhibitor,showed a synergistic inhibitory effect on OS lung metastasis and a longer survival time.In addition,tissue microarray(TMA)of OS patients also detected much higherβ-catenin expression which indicated the role ofβ-catenin in OS development.In summary,our current study not only proved that ZnO NPs can inhibit OS metastasis by degradingβ-catenin in HIF-1α/BNIP3/LC3B-mediated mitophagy pathway,but also provided a far-reaching potential of ZnO NPs in clinical OS treatment with metastasis.

Role of the Notch Signaling Pathway in Fibrosis of Denervated Skeletal Muscle

In order to investigate the role of the Notch signaling pathway in skeletal muscle fibrosis after nerve injury, 60 Sprague-Dawley rats were selected and divided randomly into a control and two experimental groups. Group A served as controls without any treatment. Rats in groups B were injected intraperitoneally with 0.2 mL PBS and those in group C were injected intraperitoneally with 0.2 mL PBS+100 ymol/L, 0.2 mL N-[N-(3,5-difluorophenacetyl)-l-alanyl]- S-phenylglycine t-butyl ester (DAPT, a gamma-secretase inhibitor that suppresses Notch signaling) respectively, on postoperative days 1, 3, 7, 10, and 14 in a model of denervation-induced skeletal muscle fibrosis by right sciatic nerve transection. Five rats from each group were euthanized on postoperative days 1, 7, 14, and 28 to collect the right gastrocnemii, and hematoxylin and eosin (HE) staining, immunohistochemistry test, real-time PCR, and Western blotting were performed to assess connective tissue hyperplasia and fibroblast density as well as expression of Notch 1, Jagged 1, and Notch downstream molecules Hes 1 and collagen I (COL I) on day 28. There was no significant difference in HE-stained fibroblast density between group B and C on postoperative day 1. However, fibroblast density was significantly higher in group B than in group C on postoperative days 7, 14, and 28. Notch 1, Jagged 1, Hes 1, and COL I proteins in the gastrocnemius were expressed at very low levels in group A but at high levels in group B. Expression levels of these proteins were significantly lower in group C than in group B (P<0.05), but they were higher in group C than in group A (P<0.05) on postoperative day 28. We are led to conclude that locking the Notch signaling pathway inhibits fibrosis progression of denervated skeletal muscle. Thus, it may be a new approach for treatment of fibrosis of denervated skeletal muscle.

Strawberry Notch 1 (SBN01) promotes proliferation of spermatogonial stem cells via the noncanonical Wnt pathway in mice

While it is known that spermatogonial stem cells (SSCs) initiate the production of male germ cells, the mechanisms of SSC self-renewal, proliferation, and differentiation remain poorly understood. We have previously identified Strawberry Notch 1 (SBN01), a vertebrate strawberry notch family protein, in the proteome profile for mouse SSC maturation and differentiation, revealing SBN01 is associated with neonatal testicular development. To explore further the location and function of SBN01 in the testes, we performed Sbnol gene knockdown in mice to study the effects of SBN01 on neonatal testicular and SSC development. Our results revealed that SBN01 is required for neonatal testicular and SSC development in mice. Particularly, in vitro Sbnol gene knockdown with morpholino oligonucleotides caused a reduction of SSCs and inactivation of the noncanonical Wnt pathway, through Jun N-terminal kinases. Our study suggests SBN01 maintains SSCs by promoting the noncanonical Wnt pathway.

Brucine Inhibits Bone Metastasis of Breast Cancer Cells by Suppressing Jagged1/Notch1 Signaling Pathways

Objective： To examine the effects of brucine on the invasion, migration and bone resorption of receptor activator of nuclear factor-kappa B ligand（RANKL）-induced osteoclastogenesis. Methods： The osteoclastogenesis model was builded by co-culturing human breast tumor MDA-MB-231 and mouse RAW264.7 macrophages cells. RANKL（50 ng/m L） and macrophage-colony stimulating factor（50 ng/m L） were added to this system, followed by treatment with brucine（0.02, 0.04 and 0.08 mmol/L）, or 10 μmol/L zoledronic acid as positive control. The migration and bone resorption were measured by transwell assay and in vitro bone resorption assay. The protein expressions of Jagged1 and Notch1 were investigated by Western blot. The expressions of transforming growth factor-β1（TGF-β1）, nuclear factor-kappa B（NF-κB） and Hes1 were determined by enzyme-linked immunosorbent assay. Results： Compared with the model group, brucine led to a dose-dependent decrease on migration of MDA-MB-231 cells, inhibited RANKL-induced osteoclastogenesis and bone resorption of RAW264.7 cells（P 〈0.01）. Furthermore, brucine decreased the protein levels of Jagged1 and Notch1 in MDA-MB-231 cells and RAW264.7 cells co-cultured system as well as the expressions of TGF-β1, NF-κB and Hes1（P〈0.05 or P〈0.01）. Conclusion： Brucine may inhibit osteoclastogenesis by suppressing Jagged1/Notch1 signaling pathways.

ZBP-89通过HIF-1α/Notch1通路对肝癌干细胞干性的调控作用

目的探究ZBP-89(Zinc-binding protein-89)通过HIF-1α/Notch1通路对肝癌干细胞干性的调控作用。方法构建ZBP-89过表达慢病毒载体(Len-ZBP-89)并进行慢病毒包装,微球体培养法富集肝癌干细胞,流式细胞术检测细胞中EpCAM、CD133的表达,CoCl 2(HIF-1α激活剂)处理肝癌干细胞,RT-PCR检测细胞中ZBP-89、 HIF-1α和Notch1mRNA表达,Westernblot检测细胞中ZBP-89、 HIF-1α、 Notch1、 CD44、CD133和EpCAM蛋白表达,Transwell检测细胞迁移和侵袭能力,光学显微镜观察特征性肿瘤球体的形成。结果 Len-ZBP-89慢病毒包装成功,病毒滴度为1.32×10^(9) TU/mL;肝癌细胞PLC/PRF/5中ZBP-89 mRNA和蛋白表达水平下调,HIF-1α和Notch1 mRNA和蛋白表达水平上调;PLC/PRF/5细胞成功富集肝癌干细胞PLC/PRF/5.C,PLC/PRF/5.C中EpCAM、CD133的表达水平均上调;Len-ZBP-89可下调PLC/PRF/5.C细胞中HIF-1α、Notch1、CD133、CD44和EpCAM的蛋白表达,下调细胞迁移、侵袭能力和特征性肿瘤球体克隆数量;CoCl 2(HIF-1α激活剂)可上调PLC/PRF/5.C细胞中HIF-1α、Notch1、CD133、CD44和EpCAM的蛋白表达,上调细胞迁移、侵袭能力和特征性肿瘤球体克隆数量(P<0.05);CoCl 2处理可逆转Len-ZBP-89对PLC/PRF/5.C干性的抑制作用。结论过表达ZBP-89通过抑制HIF-1α/Notch1通路抑制肝癌干细胞干性。

Solanine Interferes with AKT/p-AKT and PI3K/p-PI3K Pathway to Inhibit HIF and Destroy Cell Energy Metabolism

The purpose of this study was to explore the mechanism of Solanine disrupting energy metabolism in human renal cancer ACHN cells and to clarify its target. The specific method was to culture human renal cancer ACHN cell lines, and to intervene with Solanine of high, medium and low concentrations. The content of ATP in cells was measured by ELISA method. The expression of HIF-1α protein and the expression of PI3K, AKT, p-PI3K, p-AKT in PI3K/AKT pathway were detected by Western blotting. The results showed that compared with the control group, the relative expression of p-PI3K and p-AKT showed a downward trend with the increase of Solanine concentration (P < 0.05), while the relative expression of PI3K and AKT showed no significant change (P > 0.05). In addition, the relative expression of HIF-1α also showed a downward trend (P < 0.05). According to the above results, it is suggested that Solanine can significantly inhibit the energy metabolism of renal cancer cells, the main mechanism of which is the down-regulation of HI-1αf downstream of the PI3K/Akt pathway by inhibiting the phosphorylation process of PI3K/p-PI3K and Akt/p-Akt.

不同氧浓度下Notch1-4受体亚型在肺腺癌细胞A549中的表达与机制

目的探讨不同氧浓度下Notch1-4受体亚型在肺腺癌细胞A549中的表达及其机制。方法分别在正常氧浓度和低氧条件下培养肺腺癌A549细胞,采用Western blotting及RT-PCR方法分别检测Notch1-4、HIF-1α蛋白和mRNA表达。结果 Notch1-4受体各亚型在A549细胞中均有表达,Notch1蛋白在低氧浓度较正常氧浓度下表达偏高,差异有统计学意义(P <0. 05)。Notch2、3和4蛋白在不同氧浓度下表达比较,差异均无统计学意义(P> 0. 05)。Notch1-4受体各亚型在A549细胞中均有表达,Notch1 mRNA在低氧浓度较正常氧浓度表达偏高(P <0. 05),Notch2、3和4 mRNA不同氧浓度下表达比较,差异均无统计学意义(P> 0. 05)。HIF-1αmRNA及蛋白在1%O2及21%O2环境下在A549细胞中均有表达,且在低氧浓度较正常氧浓度下表达偏高,差异均有统计学意义(均P <0. 05)。结论低氧条件下,Notch1和HIF-1α在A549细胞中高表达,HIF-1α可能通过调节Notch1参与肺腺癌增殖。

γ分泌酶抑制剂阻断Notch1通路对乏氧环境骨肉瘤细胞株的影响

骨肉瘤是最常见的恶性骨肿瘤,恶性程度高,复发或转移是骨肉瘤患者死亡的最主要原因[1].肿瘤组织具有缺血缺氧的病理生理特点,目前普遍认为肿瘤缺氧与其侵袭转移特性密切相关[2].缺氧诱导因子(HIF)在肿瘤缺氧应答体系中居于核心调控位置[3],而针对细胞缺氧反应最重要的介质是HIF-1α.研究证实骨肉瘤细胞的增殖及分化主要由Notch1信号通路激活导致[4].乏氧微环境下转录调控因子HIF-1αNotch1信号通路能够诱导肿瘤细胞增殖及侵袭力增强等[5].本研究旨在观察乏氧微环境下阻断Notch1通路对骨肉瘤细胞生物学行为的影响.

Novel molecular targets in hepatocellular carcinoma

Globally,hepatocellular carcinoma(HCC)is a leading cause of cancer and cancerrelated deaths.The therapeutic efficacy of locoregional and systemic treatment in patients with advanced HCC remains low,which results in a poor prognosis.The development of sorafenib for the treatment of HCC has resulted in a new era of molecular targeted therapy for this disease.However,the median overall survival was reported to be barely higher in the sorafenib treatment group than in the control group.Hence,in this review we describe the importance of developing more effective targeted therapies for the management of advanced HCC.Recent investigations of molecular signaling pathways in several cancers have provided some insights into developing molecular therapies that target critical members of these signaling pathways.Proteins involved in the Hedgehog and Notch signaling pathways,Polo-like kinase 1,arginine,histone deacetylases and Glypican-3 can be potential targets in the treatment of HCC.Monotherapy has limited therapeutic efficacy due to the development of inhibitory feedback mechanisms and induction of chemoresistance.Thus,emphasis is now on the development of personalized and combination molecular targeted therapies that can serve as ideal therapeutic strategies for improved management of HCC.