缺氧诱导因子-1α与胰腺癌发生发层的关系

缺氧是实体瘤生长中存在的普遍现象,它和肿瘤的发展、侵润、转移密切相关。研究发现在胰腺癌组织中也存在缺氧现象。缺氧诱导因子(hypoxia-inducible factor-1,HIF-1)是缺氧条件下广泛存在于哺乳动物及人体中的一种转录因子,它是由α亚基和β亚基组成的异源二聚体。HIF-1α是HIF-1的活性部分,其表达与胰腺癌的血管生成、凋亡抑制、多药耐药、生长转移具有密切关系。同时,缺氧状态下,间质细胞与胰腺癌细胞之间的相互作用促进了癌细胞的侵袭力。现就HIF-1α在胰腺癌组织中的表达及作用做一综述。     

miR-1271-5p靶向PDK1促进胃癌细胞凋亡

目的 探讨miR-1271-5p在胃癌的作用和可能的作用机制。方法 RT-qPCR和原位杂交法检测胃癌组织和胃癌细胞株中miR-1271-5p的表达。Lipofectamine 2000转染miR-1271-5p mimics后,噻唑蓝（MTT）法和台盼蓝染色法检测SGC-790细胞的活性,Annexin V/PI染色检测细胞凋亡,JC-1探针检测线粒体膜电位,Western blot检测PDK1/Akt/凋亡信号相关蛋白的表达。荧光素酶法以及功能修复实验评估miR-1271-5p与PDK1的靶向关系。结果 胃癌组织和细胞中miR-1271-5p的表达降低。转染miR-1271-5p mimics后,SGC-790细胞的存活率下降,凋亡率上升,线粒体膜电位以及Bcl-2和p-AKT表达降低,Bax和Cleaved caspase-3表达增高。PDK1为miR-1271-5p的靶基因,过表达PDK1能逆转miR-1271-5p对胃癌细胞的促凋亡作用。结论 过表达miR-1271-5p可促进胃癌细胞凋亡,其机制可能与其靶向PDK1进而抑制AKT信号活性有关。     

缺氧诱导因子-1α表达在胃癌中的研究进展

胃癌是人类最常见的恶性肿瘤之一,严重威胁着人类的健康。大量研究表明,缺氧能促进恶性肿瘤的发展,增强其侵袭性,而缺氧诱导因子-1α(HIF-1α)是这一过程的主要调节因子,是缺氧条件下广泛存在于哺乳动物及人体内的一种转录因子,通过增加多种转录因子和靶基因产物的表达,使肿瘤在缺氧的环境下生长、增殖、侵袭及转移。本文就HIF-1α在胃癌领域的研究进展做一综述。     

高氧对雄激素敏感的前列腺癌细胞移植瘤生长及其缺氧诱导因子-1α表达的影响

目的:探讨高氧对雄激素敏感的前列腺癌细胞移植瘤生长及其缺氧诱导因子-1α表达的影响。方法:将前列腺癌前列腺淋巴结癌(LNCa P)细胞接种于36只Foxn1小鼠的双侧腹,并将其随机放置于含氧量不同的气室中并分组如下:缺氧组11例,常氧组16例,高氧组9例。处理28天后进行称重,麻醉处死,从左心室取血样;分离出移植瘤并进行称重。采用Western blotting、免疫荧光分析、血红蛋白测定的方法对各组移植瘤生长、血管生成及血管化、缺氧诱导因子-1(HIF-1α)表达以及细胞信号转导因子表达进行检测。结果:缺氧组的移植瘤生长较常氧组快(P0.05);高氧组移植瘤生长与常氧组相比差异不具有统计学意义(P0.05)。高氧组移植瘤的HIF-1α、血管内皮生长因子(VEGF)和血管内皮生长因子受体2(VEGF-R2)表达均较常氧组高,而缺氧组移植瘤的HIF-1α表达与常氧组基本相似。缺氧组移植瘤的血[HB]增长率(175%)高于常氧组(45%)。高氧组的Nrf2的表达水平较常氧组明显增加(P0.05)。结论:体内高氧诱导HIF-1α在LNCa P肿瘤高表达的同时,不会加快肿瘤的生长。     

miR-9-5p靶向Ambra1促进舌癌细胞增殖

miRNAs在肿瘤中异常表达,且与肿瘤的发生发展密切相关。目前发现,miR-9-5p在肿瘤中可能发挥原癌或抑癌效应,功能尚未完全阐述清楚。本文拟探讨miR-9-5p在舌癌中的作用。前期研究中收集10例舌癌组织及配对的癌旁组织,实时荧光定量PCR技术检测后发现,miR-9-5p在舌癌组织中的表达量显著高于癌旁组织,且其在舌癌细胞中的表达量也明显高于正常舌上皮细胞。此外,在舌癌细胞Tca8113中过表达miR-9-5p显著增加细胞的增殖能力。生物信息学预测及双荧光素酶报告基因实验证实,miR-9-5p可直接结合在自噬/苄氯素1调节因子1（activating molecule in beclin1-regulated autophagy, Ambra1）的 3′-UTR区域,靶向抑制Ambra1表达。Western印迹结果证实过表达miR-9-5p降低Ambra1的表达,反之亦然。Ambra1在舌癌细胞中的表达量显著低于正常舌上皮细胞。BrdU实验证实在舌癌细胞SCC-25中过表达Ambra1可显著抑制其增殖能力;相反,使用siRNA技术沉默Ambra1能够显著促进Tca8113细胞的增殖。在干预miR-9-5p的细胞中同时干预Ambra1的表达,结果发现Ambra1可显著逆转miR-9-5p对舌癌细胞增殖的促进作用。总之,miR-9-5p在舌癌中可能发挥原癌基因样作用,通过直接靶向抑制Ambra1表达进而促进舌癌细胞发生增殖。     

miR-153-5p靶向SNAI1介导结直肠癌细胞的放疗抵抗

目的 探讨miR-153-5p在结直肠癌放疗抵抗中的作用,并进一步研究其潜在分子机制。方法 首先对放疗敏感细胞系SW480及放疗抵抗结直肠癌细胞系SW480R中miR-153-5p表达水平进行RT-qPCR检测;然后利用转染技术构建过表达miR-153-5p的SW480R细胞株,检测过表达miR-153后SW480R在接受放射后其细胞活力、侵袭能力、集落形成能力、凋亡水平的改变;免疫组织化学染色观察放疗敏感及放疗抵抗结直肠癌组织中SNAI1的表达差异;TargetScan分析miR-153-5p潜在靶点并利用荧光素酶实验验证;检测过表达miR-153-5p及SNAI1后SW480R细胞侵袭能力、集落形成能力及凋亡水平。结果 与SW480组相比,SW480R组细胞中miR-153-5p表达降低;过表达miR-153-5p的SW480R细胞细胞活力、侵袭能力、集落形成能力均明显减弱,而凋亡水平显著增加;与放疗敏感结直肠癌组织相比,放疗抵抗结直肠癌组织中SNAI1显著增加;SNAI1可能为miR-153-5p的作用靶点;过表达miR-153-5p及SNAI1后SW480R细胞侵袭能力、集落形...     

缺氧诱导因子1

缺氧诱导因子1是缺氧诱导细胞所产生的一种蛋白质,由一个120 ku的α亚基和一个91～94 ku的β亚基组成的异源二聚体.在缺氧条件下,促进红细胞生成素和糖酵解酶等基因的转录和表达,维持机体氧稳态.     

基于生物信息学分析研究miR-182-5p通过靶向调控EP300促进乳腺癌细胞的侵袭和转移

近期研究表明,miR-182-5p对多种癌症的侵袭和转移具有重要作用,但其在乳腺癌侵袭转移中的研究相对较少。本研究通过网上在线microRNA分析工具下载乳腺癌组织及正常乳腺组织表达比较的数据集,分析发现在GSE4589、GSE38167、GSE61438等3个数据库中,在乳腺癌组织中存在26个相同的microRNA,其中8个上调,而我们实验验证发现hsa-miR-182在8例病理组织中的表达上调差异最显著(P=0.001),选定目的基因hsa-miR-182;qRT-PCR检测细胞中miR-182-5p的表达,结果显示,与MCF-10A相比,miR-182-5p在MDA-MB-231、T47D、MDA-MB-453、MCF-7中表达上调(P<0.05);转染miR-182-5p干扰质粒,qRT-PCR检测细胞中miR-182-5p的表达情况。结果显示,miR-182-5p表达显著降低(P=0.003),提示转染成功;Transwell侵袭结果显示,MDAMB-231细胞敲低miR-182-5p,与对照组相比,体外侵袭能力明显降低(P=0.002);Western印迹检测转染miR-182-5p干扰质粒时,MDA-MB-231中上皮-间质转化(epithelial-mesenchymal transition,EMT)相关标志物的表达情况,结果显示,与对照组相比,敲低miR-182-5p使细胞中上皮-钙黏着蛋白(E-cadherin)表达上调,神经-钙黏着蛋白(N-cadherin)、波形蛋白(vimentin)表达下调。为研究探讨miR-182-5p的靶蛋白,采用在线预测软件预测可能与miR-182-5p结合的靶蛋白,cytoscape构建蛋白质互作网络图并筛选出hub基因;双荧光素酶结果证实,miR-182-5p可与EP300靶向结合(P=0.001);采用qRT-PCR、Western印迹检测转染miR-182-5p干扰质粒后EP300在mRNA及蛋白质水平的表达,结果显示,与对照组相比,在敲低miR-182-5p组中EP300在mRNA及蛋白质的表达上调(P=0.001)。综上所述,miR-182-5p可靶向调节EP300,促进乳腺癌细胞的侵袭与转移。     

miR-190-5p靶向抑制PHLPP1的表达促进肺腺癌A549细胞增殖和转移

miR-190-5p是小分子非编码RNA,参与调控肺腺癌细胞的增殖和转移。对miR-190-5p调控肺腺癌细胞的增殖和转移的机制深入研究会帮助了解到肺腺癌的发生机制和探究潜在的药物干预靶点。本文研究了miR-190-5p对肺腺癌A549细胞的增殖和转移的影响,解释了miR-190-5p促进肺腺癌生长的机制;通过感染miR-190-5p的慢病毒表达系统,克隆了稳定过表达miR-190-5p的肺腺癌A549细胞系;通过生物信息学方法和荧光素酶报告实验确认了miR-190-5p的靶基因。结果发现,稳定过度表达的miR-190-5p能促进A549细胞的增殖和转移;确定了miR-190-5p的靶点是PH域富含亮氨酸重复的蛋白磷酸酶1(Leucine-rich repeat-rich protein phosphatase 1 in the PH domain,PHLPP1),miR-190-5p降低PHLPP1蛋白水平;稳定表达的miR-190-5p增加了A549细胞中E盒结合锌指蛋白1(E box-binding zinc finger protein1,TCF8/ZEB1)、锌指转录因子(Z...     

缺氧诱导因子-1α在缺氧预处理预防心肌细胞损伤中的作用

本工作旨在研究缺氧预处理(hypoxic preconditioning,HPC)对于心肌细胞外信号调节激酶(extracellular signal-regulated proteinkinases,ERK)活性、缺氧诱导因子-1α(hypoxia-inducible factor-1α,HIF-1α)表达的影响,及其在缺氧复氧诱导心肌细胞损伤中的作用。通过在培养的SD乳鼠心肌细胞缺氧／复氧(H／R)模型上,观察HPC对于24h后H／R诱导心肌细胞损伤的影响,以台盼蓝排斥实验检测心肌细胞存活率、以TUNEL法检测细胞凋亡、并用荧光素染料Hoechst33258测定心肌细胞凋亡率：制备心肌细胞蛋白提取物,以磷酸化的ERK1／2抗体测定ERK1／2活性,以抗HIF-1α抗体检测HIF-1α的表达,并观察ERKs的上游激酶(MEK1／2)抑制剂PD98059对于HPC诱导的ERKs磷酸化、HIF-1α表达以及心肌细胞保护作用的影响,并分析细胞损伤与ERK1／2活性、HIF-1α表达量之间的相互关系。结果 显示缺氧复氧造成心肌细胞损伤,HPC可以增加心肌细胞H／R后存活率,降低凋亡率,并激活ERKll2,诱导HIF-1α表达：细胞凋亡与ERKs活性、HIF-1α表达量之间存在负相关,即ERKs活化、HIF-1α表达与预防细胞损伤有关：而ERKs活性与HIF-1α表达量之间存在正相关,ERKs的上游激酶MEK抑制剂PD98059可以消除HPC诱导的ERKs磷酸化、HIF-1α表达和心肌细胞保护作用。由此得出的结论是HPC可以提高乳鼠心肌细胞对于H／R的耐受性,其机制涉及ERKs介导的HIF-1α表达。     

Circ_0005615 promotes cervical cancer cell growth and metastasis by modulating the miR-138-5p/KDM2A axis

Cervical cancer (CC) is a highly fatal gynecological malignancy due to its high metastasis and recurrence rate. Circular RNA (circRNA) has been regarded as a regulator of CC. However, the underlying molecular mechanism of circ_0005615 in CC remains unclear. The levels of circ_0005615, miR-138-5p, and lysine demethylase 2A (KDM2A) were measured using qRT-PCR or western blot. Cell proliferation was assessed by Cell Counting Kit-8, 5-ethynyl-2′-deoxyuridine, and colony formation experiments. Cell invasion and migration were tested by transwell assay and wound healing assay. Flow cytometry and Caspase-Glo 3/7 Assay kit were used to analyze cell apoptosis. The expression of proliferation-related and apoptosis-related markers was detected by western blot. The binding relationships among circ_0005615, miR-138-5p, and KDM2A were verified by dual-luciferase reporter assay or RNA immunoprecipitation assay. Xenograft assay was applied to detect the effect of circ_0005615 in vivo. Circ_0005615 and KDM2A were upregulated, while miR-138-5p was downregulated in CC tissues and cells. Circ_0005615 knockdown retarded cell proliferation, migration, and invasion, while promoting apoptosis. Besides, circ_0005615 sponged miR-138-5p, and miR-138-5p could target KDM2A. miR-138-5p inhibitor reversed the regulation of circ_0005615 knockdown on CC cell growth and metastasis, and KDM2A overexpression also abolished the inhibitory effect of miR-138-5p on CC cell growth and metastasis. In addition, we also discovered that circ_0005615 silencing inhibited CC tumor growth in vivo. Circ_0005615 acted as a tumor promoter in CC by regulating the miR-138-5p/KDM2A pathway.     

miR-17-5p promotes proliferation and epithelial-mesenchymal transition in human osteosarcoma cells by targeting SRC kinase signaling inhibitor 1

MicroRNA-17-5p (miR-17-5p) and epithelial-mesenchymal transition (EMT) have been reported to participate in the development and progression of multiple cancers. However, the relationship between the miR-17-5p and EMT in osteosarcoma (OS) is still poorly understood. This study was to investigate the effects of the miR-17-5p and its potential mechanism in regulating proliferation, apoptosis, and EMT of human OS. Quantitative real-time PCR was used to detect the miR-17-5p and SRC kinase signaling inhibitor 1 (SRCIN1) messenger RNA expression in OS specimens and cell lines. After transfection with miR-17-5p inhibitors, proliferation, apoptosis, migration, and invasion of OS cells were assessed by using the Cell Counting Kit-8, the annexin V-FITC apoptosis, wound-healing, and transwell assays. The SRCIN1 was validated as a target of the miR-17-5p through bioinformatics algorithms and luciferase reporter assay. Moreover, the expression of EMT markers, E-cadherin, N-cadherin, and Snail was identified by the Western blot analysis. MiR-17-5p was significantly upregulated in OS tumor samples and cell lines. It inhibited proliferation and EMT, and promoted apoptosis in OS. The SRCIN1 was identified as a direct target of the miR-17-5p. Silenced miR-17-5p could change the expression of EMT markers, such as upregulating the expression of E-cadherin, and downregulating the expression of N-cadherin and Snail through targeting the antioncogenic SRCIN1. These findings suggest that the miR-17-5p promotes cell proliferation, and EMT in human OS by directly targeting the SRCIN1, and reveal a branch of the miR-17-5p/SRCIN1/EMT signaling pathway involved in the progression of OS.     

miR-129-5p and -3p co-target WWP1 to suppress gastric cancer proliferation and migration

Gastric cancer (GC) is a worldwide health problem. Uncovering the underlining molecular mechanisms of GC is of vital significance. Here, we identified a novel oncogene WW domain-containing E3 ubiquitin protein ligase 1 (WWP1) in GC. WWP1 could promote GC cell proliferation and migration in vitro and expedite GC growth in vivo. We also found out two microRNAs (miRNAs): miR-129-5p and -3p could both target WWP1. Interestingly, miR-129-5p bound to the CDS region of WWP1 mRNA. The miR-129 pairs (miR-129-5p and -3p) play pivotal roles in GC to suppress its proliferation and migration in vitro and slow down GC growth in vivo by repressing WWP1. In summary, we identified two tumor suppressive miRNAs which share the same precursor that could regulate the same oncogene WWP1 in GC. Our finding would add new route for GC research and treatment.     

miR-138-5p targets MCU to inhibit mitochondrial biogenesis and colorectal cancer growth

miR-138-5p has been identified as a novel cancer-related miRNA molecule in a variety of malignancies. However, the functions and mechanisms underlying miR-138-5p in colorectal carcinoma (CRC) remains largely unknown. In the present study, we analysed the biological effects and clinical significance of miR-138-5p in CRC. miR-138-5p expression was analysed by quantitative real-time PCR in CRC tissues and cell lines. The effects of miR-138-5p on CRC cell growth was detected by cell proliferation, colony formation, cell cycle and cell apoptosis assays in vitro and in vivo. Our data showed that miR-138-5p was significantly downregulated in CRC. Downregulated miR-138-5p was related with poor prognosis in patients with CRC. miR-138-5p suppressed CRC growth but promoted cell death both in vitro and in vivo. Online predictions and integrated experiments identified that miR-138-5p targeted MCU, and downregulated miR-138-5p promoted mitochondrial biogenesis in CRC. In the light of the underlying mechanisms, our results indicated that downregulated miR-138-5p led to increased expression of MCU, which subsequently increased the production of ROS to promote CRC growth. Our results indicated that downregulated miR-138-5p strengthened mitochondrial biogenesis through targeting MCU, thus contributing to CRC cell growth, which may provide a potential therapeutic target for CRC.     

miR-548c-5p inhibits colorectal cancer cell proliferation by targeting PGK1

Accumulating studies have implicated that microRNAs (miRNAs) are involved in the pathogenesis of colorectal cancer (CRC). However, the role of miR-548c-5p, a novel identified miRNA in malignancies, in colorectal carcinogenesis remains largely unknown. The present study is aimed to investigate the effect and molecular mechanism of miR-548c-5p in CRC by a sequence of cellular experiments. miR-548c-5p was significantly downregulated, whereas phosphoglycerate kinase 1 (PGK1), a key enzyme for glycolysis, was obviously upregulated in peripheral blood mononuclear cells and cancer tissues from patients with CRC. Besides, miR-548c-5p and PGK1 were negatively associated with each other. The luciferase reporter assay revealed that PGK1 was a targeted gene of miR-548c-5p. Moreover, the proliferation and generation of inflammatory cytokines (TNF-α and IL-6) were significantly inhibited in miR-548c-5p-overexpressed SW480 CRC cells stimulated by lipopolysaccharide (LPS). Accordingly, miR-548c-5p may serve as a cancer suppressor in CRC by targeting PGK1.     

microRNA-1225 inhibit apoptosis of pancreatic cancer cells via targeting JAK1

The microRNA miRNA-1225-5p (miR-1225) is known as an essential modulator of the development of multiple cancers and other biological reactions. However, the understanding of its contribution to pancreatic cancer (PC) is insufficient. The effects of miR-1225 on PC cell survival and tumorigenesis in vivo as well as on the modulation of cell apoptosis were investigated. The expression of miR-1225 was upregulated in 20 human LC samples from acute myeloid leukemia patients with adverse prognosis and poor responses to therapy as well as in several human PC cell lines, as compared to that in healthy tissues, normal tissues, and normal pancreatic cells. In contrast, Janus kinase 1 (JAK1) expression was downregulated in human-derived PC samples and PC cell lines. EdU staining demonstrated that the aberrant expression of miR-1225 impaired the proliferation and survival of these two PC cell lines. The depletion of miR-1225 expression increased the apoptosis of both PANC-1 and AsPC-1 cells, as revealed by the TdT-mediated dUTP nick end labeling (TUNEL) staining and flow cytometry results. The results of dual-luciferase reporter assay indicated that miR-1225 targeted the 3′-untranslated region of JAK1 for silencing. Silencing of JAK1 expression counteracted the suppressive influence of miR-1225 depletion in PC cells. Thus, these results offer an insight into the biological and molecular mechanisms underlying the development of PC and provide potential strategies for PC treatment.