BMP9下调HIF-1α抑制乳腺癌MDA-MB-231细胞的有氧糖酵解和迁移侵袭

目的 研究骨形成蛋白BMP9对三阴性乳腺癌MDAMB-231细胞有氧糖酵解和迁移侵袭的调控作用。方法 实验组使用人BMP9重组腺病毒（AdBMP9）感染MDA-MB-231细胞,对照组用空载的GFP腺病毒感染细胞。采用乳酸、葡萄糖和ATP检测试剂盒检测细胞的葡萄糖摄取量、乳酸和ATP生成量;通过GEPIA2数据库,分析BMP9在泛癌中与糖酵解关键酶基因的相关性;qRT-PCR检测过表达BMP9后,MDA-MB-231中糖酵解关键酶GLUT1、HK2、PKM2、LDHA的mRNA表达水平;STRING数据库分析BMP9抑制MDA-MB-231有氧糖酵解潜在靶点;Western blot检测细胞HIF-1α和下游蛋白表达水平;划痕实验和Transwell实验评估不同处理后,细胞的迁移与侵袭能力的改变。结果 与对照组相比,BMP9下调乳腺癌MDA-MB-231细胞的葡萄糖摄取、乳酸生成及ATP水平（P<0.01）,抑制HIF-1α及其下游蛋白表达;Rescue实验中,过表达HIF-1α能逆转BMP9对MDAMB-231细胞有氧糖酵解和迁移侵袭的抑制作用。结论 BMP9下调HIF-1α抑...     

靶向缺氧诱导因子-1的抗肿瘤药物研究进展

缺氧诱导因子-1(hypoxia inducible factor-1，HIF-1)是调控肿瘤内缺氧效应的一个重要因子，它能引起肿瘤细胞的无氧糖酵解、引发肿瘤的血管生成、促进肿瘤细胞的增殖、侵袭及迁移等行为，使这些肿瘤细胞在缺氧微环境下得以存活并使肿瘤的恶性程度进一步增强。它还能引发肿瘤对放/化疗的耐受，其表达程度与预后不良成正相关。它的发现为人们靶向肿瘤缺氧来开发新型抗肿瘤药物提供了一个潜在的分子靶点。本文综述了近年来靶向缺氧诱导因子HIF-1的抗肿瘤药物研究进展。     

低氧诱导因子-1α对细胞信号、细胞凋亡的影响及研究进展

低氧诱导因子-1（hypoxia—induciblefactor-1，HIF-1）是普遍存在于人和哺乳动物细胞内的缺氧应答调控因子，通过调控一系列与缺氧适应有关基因的表达以保持机体的氧稳态。HIF-1（尤其是HIF-1α）的表达水平与糖酵解、细胞周期阻滞、细胞生存与增殖、血管新生、血管舒缩、红细胞生成、肿瘤生长和转移以及肿瘤多药耐药等许多生命活动密切相关。多数肿瘤具有缺氧的微环境。HIF-1α对于肿瘤的效应是多方面的：HIF-1α在上游调控诸多与肿瘤发生密切相关的基因；肿瘤在低氧条件下选择更恶性的表型；增加细胞突变率；增加与血管新生和肿瘤侵袭有关的基因表达；低氧条件下细胞更具有耐药倾向。因此HIF-1α已经成为一个治疗肿瘤的新的关键性靶点。细胞信号、细胞凋亡在多细胞生物的个体发育、自稳平衡等生理以及肿瘤、炎症等病理过程中具有重要意义，理解HIF-1α对于细胞信号、细胞凋亡的调节，对于开发特异性的肿瘤治疗有着现实意义。现就HIF-1α对细胞信号、细胞凋亡的影响及研究进展作一综述。     

乌梅丸含药血清对胰腺癌细胞增殖及糖酵解的影响

摘要：目的:探讨乌梅丸含药血清对人胰腺癌SW1990细胞增殖及糖酵解的影响。方法:制备乌梅丸含药血清,MTT法检测乌梅丸含药血清对SW1990细胞增殖的影响,并筛选最佳含药血清浓度;检测不同作用时间（0,12,24 h）乌梅丸含药血清对SW1990细胞乳酸累积的影响及细胞中缺氧诱导因子1α（HIF-1α）、6-磷酸果糖-2-激酶/果糖-2,6-二磷酸酶3（PFKFB3）蛋白的表达水平,初步探索乌梅丸含药血清影响胰腺癌细胞增殖与胰腺癌糖酵解的关系。结果:乌梅丸含药血清可明显抑制人胰腺癌SW1990细胞的增殖活性,并且从作用12 h后乳酸累积量明显降低（P<0.05或P<0.01）,糖酵解反应相关HIF-1α、PFKFB3蛋白表达水平显著下降（P<0.01）。结论:乌梅丸可抑制胰腺癌细胞的增殖,其机制可能是通过抑制HIF-1α及糖酵解关键限速酶PFKFB3的表达,抑制肿瘤细胞糖酵解,减少乳酸堆积,改善肿瘤酸性微环境而实现的。     

肿瘤细胞的“瓦博格效应”与抗肿瘤药物的研发

“瓦博格效应”即指肿瘤细胞对糖酵解通路产能依赖增强的现象。这一现象为设计各种抗肿瘤代谢的药物提供了生物化学基础。介绍肿瘤细胞所具有的“瓦博格效应”及其主要机制，并总结了近年来基于该效应的抗肿瘤药物的研发状况。     

肿瘤代谢靶点用于癌症治疗的研究进展

肿瘤细胞可以改变糖酵解和谷氨酰胺代谢等代谢途径,产生其快速增殖所需的原料,从而使自身增殖和生存。因此,研究肿瘤代谢途径的改变,有利于找到治疗癌症疾病的靶点。本文综述了肿瘤细胞中有氧糖酵解、谷氨酰胺代谢、三羧酸循环和合成代谢的代谢特征,并详细介绍了这些代谢途径中用于癌症治疗的代谢靶点和相应的治疗药物,探讨了可以成为癌症治疗靶点的潜在标志物和肿瘤代谢靶向治疗所面临的挑战。     

缺氧对食管癌细胞缺氧诱导因子-1α及己糖激酶表达的影响

目的探讨不同缺氧程度对食管癌细胞株TE1中缺氧诱导因子-1α（Hypoxia-inducible factor-1α,HIF-1α）及糖酵解关键酶表达的影响。方法 TE1细胞分别在正常氧分压和缺氧条件下培养,缺氧时间设定为6、12、24及48h,使用Western blot方法检测缺氧培养不同时间后细胞中HIF-1α及糖酵解关键酶己糖激酶II（Hexokinase-II,HK-II）的蛋白水平表达变化。结果常氧及缺氧条件下TE1细胞中HIF-1α及HK-II均有表达,缺氧后表达均较常氧培养时明显增强,且随缺氧时间不同而呈先增多后减少的动态变化。结论低氧能够增加食管癌细胞中HIF-1α及HK-II表达而促进糖酵解进程,联合抑制HIF-1α和糖酵解酶可能成为治疗食管癌的潜在的靶点。     

糖代谢重编程及其靶向治疗药物在炎症相关疾病中的作用

细胞在炎性微环境影响下发生糖代谢重编程,使其主要供能方式由氧化磷酸化转变为有氧糖酵解,该过程参与炎症相关疾病发生发展的各个阶段。糖代谢重编程不仅改变单个细胞的代谢模式,而且打破了机体微环境的代谢稳态,进一步促进细胞有氧糖酵解,为炎症相关疾病的恶性进展提供有利条件。有氧糖酵解的相关代谢酶、转运蛋白、代谢产物等均为关键信号分子,药物通过靶向这些特异性强的关键分子,抑制有氧糖酵解从而发挥治疗作用。本文围绕糖代谢重编程对炎症相关肿瘤、类风湿关节炎、阿尔茨海默病等炎症相关疾病发生发展的影响以及药物靶向糖代谢重编程对疾病的治疗作用展开综述。     

乳酸脱氢酶A在消化系统肿瘤中的作用及相关药物研究进展

消化系统恶性肿瘤是严重威胁世界各地人类健康的高发恶性肿瘤，其目前传统治疗方式的疗效及预后尚无法达到预期，因此亟须寻找肿瘤治疗新靶点，实现肿瘤的靶向治疗手段。肿瘤细胞的能量代谢异常被视为癌症的标志，恶性肿瘤细胞通过有氧糖酵解途径摄取葡萄糖，在一系列酶的催化下获取少量能量并生成乳酸。乳酸脱氢酶A(lactate dehydrogenase A,LDHA)作为肿瘤细胞有氧糖酵解途径中的关键酶，在肿瘤细胞的代谢改变中扮演着重要角色，研究证明LDHA在多种肿瘤细胞中具有高表达特性，在临床上其高表达常与肿瘤的预后不良和高转移率有关，有望成为肿瘤治疗新靶点。本文综述了LDHA在消化系统肿瘤发生发展中的作用及其相关药物研究进展。     

Egl-9家族缺氧诱导因子1的研究进展及与肺癌的关系

氧敏感因子Egl-9家族缺氧诱导因子1(EGLN1)为哺乳动物体内重要的氧传感器,通过编码脯氨酰羟化酶结构域（PHD）中的PHD2,羟基化缺氧诱导因子-1α(HIF-1α),促进HIF-1α蛋白酶体的降解,从而影响机体的红细胞生成、糖酵解、铁代谢等。EGLN1与红细胞增多症、心脏疾病、脑代谢疾病等多种疾病相关,也可通过抑制肿瘤血管生成、与抑癌基因相互作用等影响肿瘤的发生与发展,但其在不同癌症中具有不同的作用。肺癌作为最常见和恶性程度最高的肿瘤,与EGLN1的关系也越来越受到人们的关注,但其具体机制尚不完全清楚。本文系统阐述EGLN1的结构及作用机制、在肿瘤中的表达及意义以及与肺癌的关系,旨在对EGLN1基因的研究进展及与肺癌的关系进行进一步探讨,从而为肺癌的靶向治疗提供新的思路。     

Ginsenoside 20(S)-Rg3 upregulates HIF-1α-targeting miR-519a-5p to inhibit the Warburg effect in ovarian cancer cells

The Warburg effect, one of the metabolic hallmarks of cancer, is responsible for rapid energy production through a high rate of aerobic glycolysis. Ginsenoside 20(S)-Rg3 antagonizes the Warburg effect in ovarian cancer cells by upregulating some microRNAs, including miR-519a-5p, that target key enzymes involved in aerobic glycolysis. How 20(S)-Rg3-upregulated miR-519a-5p influences the Warburg effect in ovarian cancer cells remains poorly defined, however. Here we report that while overexpression of miR-519a-5p in ovarian cancer cells inhibited the Warburg effect, inhibition of miR-519a-5p negated the suppressive action of 20(S)-Rg3 against the Warburg effect as evidenced by a decrease in glucose consumption, lactate production and HK2 expression. We identified HIF-1α as a direct target of miR-519a-5p via luciferase reporter assays and demonstrated the counteraction by overexpressed HIF-1α of 20(S)-Rg3-suppressed Warburg effect. Further, 20(S)-Rg3 decreased DNMT3A-mediated DNA methylation in the promoter region of its precursor gene, leading to an increase in the level of miR-519a-5p. In conclusion, 20(S)-Rg3 upregulates miR-519a-5p via reducing DNMT3A-mediated DNA methylation to inhibit HIF-1α-stimulated Warburg effect in ovarian cancer.     

Proteomic analysis of oral cancer reveals new potential therapeutic targets involved in the Warburg effect

Activation of peroxisome proliferator‐activated receptor alpha (PPARα) has been reported to disrupt tumour metabolism and to promote anticancer activity through interfering with the Warburg effect. This study is to investigate whether Warburg effect‐related proteins also could be identified in oral tumour lesions and to explore the functional significance of PPARα in metabolic shift. Five pairs of tongue tumour tissues and adjacent reference tissues obtained from 4‐NQO/arecoline induced mouse model were analyzed by 2‐d ‐gel‐electrophoresis and LC‐MS. Further, the hexokinase II level, pyruvate dehydrogenase (PDH) activity, and metabolites of glycolysis and TCA cycle were all examined in order to validate the effect of PPARα on metabolic shift. Changes in protein expression levels revealed that seven proteins, which were involved in glycolysis, the tricarboxylic acid cycle, and the respiratory chain, were down‐regulated in tumour tissues. We found that activation of PPARα through fenofibrate could inhibit oral cancer cell growth and switch the way of energy production from the Warburg effect to oxidative phosphorylation. Fenofibrate induced a reduction of hexokinase II protein levels, increases in PDH activity and metabolites of the TCA cycle, and an impairment of ATP production. These findings suggested that activation of the PPARα to reprogram the metabolic pathway might impair the Warburg effect and trigger cancer cell death. The study provides a novel view of changes in protein expression profiles involved in the Warburg effect during oral tumourigenesis. Activation of the PPARα to impair the Warburg effect might offer a new strategy for oral cancer treatment.     

胃癌BGC-823细胞中HIF-1α、HIF-2α对PKM2的调控作用

目的研究siRNA特异性沉默人胃癌BGC-823细胞中缺氧诱导因子(HIF-1α、HIF-2α)后PKM2的表达情况,探索HIF-1α、HIF-2α对胃癌细胞内PKM2的调控作用。方法用Realtime RT-PCR及Western blot法检测转染HIF-1α、HIF-2α及Control siRNA后细胞内PKM2表达情况。结果 RNA干扰技术能有效沉默BGC-823细胞中的HIF-1α、HIF-2α。转染HIF-1α、HIF-2αsiRNA后,PKM2在mRNA及蛋白表达水平均下降(P<0.05),其中siRNA抑制HIF-1α后,PKM2下调最明显;转染Control siRNA后,PKM2在mRNA及蛋白表达水平均无明显变化(P>0.05)。结论 HIF-1α和HIF-2α均参与调控PKM2的表达,其中HIF-1α起主要调控作用,HIF-1α、HIF-2α与PKM2共同促进肿瘤的Warburg效应。     

Therapeutic potential of targeting glucose metabolism in glioma stem cells

Glioblastoma is a highly lethal cancer. Glioma stem cells (GSCs) are potentially an attractive therapeutic target and eradication of GSCs may impact tumor growth and sensitize tumors to conventional therapies. The brain is one of the most metabolically active organs with glucose representing the most important, but not the only, source of energy and carbon. Like all other cancers, glioblastoma requires a continuous source of energy and molecular resources for new cell production with a preferential use of aerobic glycolysis, recognized as the Warburg effect. As selected metabolic nodes are amenable to therapeutic targeting, we observed that the Warburg effect may causally contribute to glioma heterogeneity. This Editorial summarizes recent studies that examine the relationship between GSCs and metabolism and briefly provides our views for the future directions. The ultimate goal is to establish a new concept by incorporating both the cellular hierarchical theory and the cellular evolution theory to explain tumor heterogeneity. Such concept may better elucidate the mechanisms of how tumors gain cellular and molecular complexity and guide us develop novel and effective targeted therapies.     

The warburg effect: why and how do cancer cells activate glycolysis in the presence of oxygen?

Cells can obtain energy through the oxygen-dependent pathway of oxidative phosphorylation (OXPHOS) and through the oxygen-independent pathway of glycolysis. Since OXPHOS is more efficient in generating ATP than glycolysis, it is recognized that the presence of oxygen results in the activation of OXPHOS and the inhibition of glycolysis (Pasteur effect). However, it has been known for many years that cancer cells and non-malignant proliferating cells can activate glycolysis in the presence of adequate oxygen levels (aerobic glycolysis or Warburg effect). Accumulating evidence suggests that the persistent activation of aerobic glycolysis in tumor cells plays a crucial role in cancer development; the inhibition of the increased glycolytic capacity of malignant cells may therefore represent a key anticancer strategy. Although some important knowledge has been gained in the last few years on this growing field of research, the basis of the Warburg effect still remains poorly understood. This communication analyzes why cancer cells switch from OXPHOS to glycolysis in the presence of adequate oxygen levels, and how these cells manage to avoid the inhibition of glycolysis induced by oxygen. Several strategies and drugs that may interfere with the glycolytic metabolism of cancer cells are also shown. This information may help develop anticancer approaches that may have clinical relevance.     

Metabolomics and cancer drug discovery: let the cells do the talking

Recent developments in cancer research have led to reconsiderations regarding metabolic dysfunctions in cancer cell proliferation and differentiation. The original concept stemmed from the observation that, even in presence of oxygen, highly proliferating cells tend to generate energy strictly from the glycolytic pathway, through a process called aerobic glycolysis, also known as the Warburg effect. More recently, advances in the field of metabolomics applied to cancer research enabled the documenting of the generality of the Warburg effect in a broad variety of tumors. Through metabolomics, cancer cells told us that oxidative stress, while representing one leading cause of genetic instability underpinning carcinogenesis, could also deliver a window of probable therapeutic opportunities that is worth opening.     

Lactate dehydrogenase downregulation mediates the inhibitory effect of diallyl trisulfide on proliferation,metastasis, and invasion in triple‐negative breast cancer

The Warburg effect plays a critical role in tumorigenesis, suggesting that specific agents targeting Warburg effect key proteins may be a promising strategy for cancer therapy. Previous studies have shown that diallyl trisulfide (DATS) inhibits proliferation of breast cancer cells by inducing apoptosis in vitro and in vivo. However, whether the Warburg effect is involved with the apoptosis‐promoting action of DATS is unclear. Here, we show that the action of DATS is associated with downregulation of lactate dehydrogenase A (LDHA), an essential protein of the Warburg effect whose upregulation is closely related to tumorigenesis. Interestingly, inhibition of the Warburg effect by DATS in breast cancer cells did not greatly affect normal cells. Furthermore, DATS inhibited growth of breast cancer cells, particularly in MDA‐MB‐231, a triple‐negative breast cancer (TNBC) cell, and reduced proliferation and migration; invasion was reversed by over‐expression of LDHA. These data suggest that DATS inhibits breast cancer growth and aggressiveness through a novel pathway targeting the key enzyme of the Warburg effect. Our study shows that LDHA downregulation is involved in the apoptotic effect of DATS on TNBC. © 2016 Wiley Periodicals, Inc. Environ Toxicol 32: 1390–1398, 2017.