PKM2对黑色素瘤细胞侵袭与迁移及巨噬细胞M2转变的影响

目的 探讨黑色素瘤中丙酮酸激酶M2(PKM2)的表达情况及其与预后相关性,阐明PKM2如何通过糖酵解途径影响黑色素瘤侵袭与迁移及对巨噬细胞M2转变的影响。方法 利用TCGA数据库及免疫组化分析PKM2在黑色素瘤组织中的表达情况;利用基因过表达、敲低及ECAR技术分析PKM2对黑色素瘤细胞糖酵解过程影响;通过划痕实验及Transwell实验验证PKM2对黑色素瘤细胞侵袭与迁移的影响;通过细胞共培养检测PKM2对巨噬细胞M1/M2转变的影响。结果 TCGA数据库显示,与正常组织相比,PKM2 mRNA水平在黑色素瘤组织中表达明显升高(P<0.05),且PKM2高表达水平患者的生存率低于低/中表达患者(P=0.001 8);免疫组化染色结果显示,PKM2在黑色素瘤发生转移患者中的评分明显高于未转移患者,差异有统计学意义(P<0.05)。敲低PKM2表达后,黑色素瘤细胞糖酵解能力、糖酵解容量和乳酸水平均明显降低(P<0.05)。细胞迁移实验结果显示,敲低PKM2表达后,黑色素瘤细胞的迁移和侵袭能力均明显降低,加入乳酸后,可部分逆转该现象。过表达组黑色素瘤巨噬细胞的肿瘤坏死因子...     

丙酮酸激酶M2型在恶性肿瘤中的表达及其临床检测研究进展

丙酮酸激酶M2型(PKM2)是肿瘤细胞有氧糖酵解途径的关键酶之一.它通过调节肿瘤细胞的代谢、基因表达和细胞周期进程等,与肿瘤的发生、发展、转移、复发等密切相关.在多种恶性肿瘤患者(如肺癌、胃癌、结直肠癌、乳腺癌等)的组织样本中发现,相较于癌旁组织,肿瘤组织中PKM2表达水平显著增高.PKM2高表达与患者较差的临床表现和...     

干扰PKM2对急性白血病细胞增殖和凋亡的影响及分子机制

目的:探讨糖酵解酶丙酮酸激酶M2型(PKM2)对人白血病细胞HL-60增殖和凋亡的影响及其分子机制。方法:以干扰PKM2的质粒转染HL-60细胞株(si-PKM2组),同时将转染空载体的HL-60细胞设为对照组(si-Ctl组)。采用RT-q PCR和Western blot技术分别检测si-Ctl组和si-PKM2组PKM2 m RNA和蛋白水平的变化;CCK-8检测两组细胞的增殖能力;采用流式细胞术检测两组细胞周期和凋亡的变化;Western blot和RT-q PCR技术检测两组细胞中PI3K/Akt/m TOR信号通路的变化及糖酵解相关基因的表达变化,并检测两组细胞中葡萄糖消耗和乳酸生成的变化情况。过表达PKM2的细胞给予PI3K抑制剂LY294002或给予半乳糖培养后检测细胞的增殖能力、周期和凋亡的变化以及葡萄糖消耗和乳酸生成的变化。结果:干扰PKM2表达后,si-PKM2组中PKM2的m RNA (t=45.21,P0.001)和蛋白水平(t=13.56,P0.01)显著降低,si-PKM2组的细胞增殖能力较对照组明显下降(F=253.59,P0.05),敲低PKM2后细胞被显著阻滞于G_1期,且明显诱导了细胞的凋亡(t=56.38,P0.05)。与对照组相比,si-PKM2组p-Akt和p-m TOR水平显著降低(t=50.23、13.51,P0.01),葡萄糖消耗和乳酸生成显著下降(t=23.16、15.20,P0.05)。补救实验结果显示,过表达PKM2给予PI3K抑制剂LY294002后减弱了其诱导的葡萄糖消耗和乳酸的产生(t=45.13、26.35,P0.05),半乳糖培养过表达PKM2的HL-60细胞对细胞增殖能力(t=16.52,P0.05)、周期及凋亡无显著影响(t=48.63,P0.05)。结论:敲低PKM2可以抑制白血病HL-60细胞的增殖并诱导其凋亡,其分子机制可能与PKM2介导的PI3K/Akt/mTOR糖酵解轴抑制有关,提示PKM2可能作为白血病防治的一个分子靶点。     

腺病毒介导p53基因对人胃癌细胞热增敏的作用

目的 评价腺病毒介导p53基因对人胃癌细胞热增敏的作用。方法 以重组腺病毒介导p53基因悬液（Adp53）感染4种不同p53状况的人胃癌细胞，用免疫组化法和Western blot法检测p53蛋白在胃癌细胞中的表达；用经胞存活分数来反映经胞增殖状况；用TUNEL法来检测细胞凋亡，感染Adp53的W和M胃癌细胞经42℃2h或43℃0.5h加温后24h，用流式细胞计检测细胞周期分布和凋亡；胃癌细胞的种植肿瘤内注射Adp53悬液后48h，行43℃0.5h加温，以肿瘤相对体积增长曲线观察肿瘤抑制情况。结果 高效靶比（100MOI）Adp53产生细胞高转染率和p53基因在4种胃癌细胞中均高表达，并产生G2/M期阻滞和凋亡及细胞增殖抑制，Adp53基因的作用不依赖胃癌细胞内在的p53状态。如果以凋亡作为热效应，Adp53对2种加温方式的热增敏比，对W细胞为1.6-3.3，而对M细胞为1.8-2.1,Adp53对W细胞肿瘤43℃0.5h加温的热增敏比为1.7，而对M细胞肿瘤为1.6。结论 腺病毒介导p53基因提高了人胃癌细胞的热敏感性，这种作用不依赖于细胞内在p53状况，本实验为p53基因治疗与热疗结合提供了可靠的实验依据。     

阿伐他汀通过PTEN/mTOR信号调节糖酵解代谢逆转白血病耐药的作用及机制

目的：探讨阿伐他汀对耐阿霉素人急性早幼粒白血病细胞系HL-60/ADM糖酵解代谢的影响及作用机制。方法：取对数生长期耐阿霉素白血病细胞HL-60/ADM，给予不同浓度阿伐他汀处理后，采用CCK-8法测定细胞增殖活性，流式细胞术检测细胞凋亡，葡萄糖消耗实验检测白血病细胞糖酵解活性，Western blot方法检测PTEN、p-m TOR、PKM2、HK2、P-gp、MRP1蛋白的表达；将PTEN-si RNA转染至HL-60/ADM细胞后，进一步采用上述方法检测PTEN低表达对阿伐他汀调节HL-60/ADM细胞凋亡及糖酵解代谢的影响。结果：CCK-8结果显示，阿伐他汀呈浓度依赖性和时间依赖性抑制HL-60/ADM细胞增殖（r=0.872,r=0.936）,10μmol/L阿伐他汀干预24 h后HL-60/ADM细胞增殖活性下降最明显，增殖活性降至（32.3±2.18）%。流式细胞术结果显示，阿伐他汀诱导HL-60/ADM细胞凋亡，该作用呈浓度依赖性（r=0.796）,10μmol/L阿伐他汀对HL-60/ADM细胞的诱导凋亡作用最强，细胞凋亡率达到（48.78±2.95）%。葡萄糖消耗实...     

Wnt信号通路抑制剂对下调PKM2表达后的肺癌A549细胞增殖凋亡及ROS水平的影响

目的研究Wnt信号通路抑制剂对下调丙酮酸激酶M2亚型(PKM2)表达后的肺癌A549细胞增殖凋亡及活性氧(ROS)水平的影响。方法肺癌A549细胞转染PKM2小干扰RNA(PKM2 siRNA组)和小干扰RNA阴性对照(siRNA-NC组),以没有转染的A549细胞作为对照组。Real-time PCR和Western blot检测三组PKM2水平。用含有5μmol/L的Wnt信号通路抑制剂IWR-1的细胞培养液培养,设置为PKM2 siRNA+IWR-1组和IWR-1组,同时以不含有IWR-1的正常细胞培养液培养PKM2 siRNA组和对照组细胞。噻唑蓝(MTT)检测细胞增殖,流式细胞术检测细胞凋亡,二氯二氢荧光素二乙酸酯(H2DCFDA)法检测ROS水平,Western blot检测β-连环蛋白(β-catenin)、c-myc、活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved Caspase-3)水平。结果细胞转染PKM2 siRNA后的PKM2 mRNA和蛋白水平、细胞存活率明显下降,细胞凋亡率、ROS水平明显升高,细胞中β-catenin、c-myc水平下降,而Cleaved Caspase-3水平升高,与不转染的对照组细胞相比差异具有统计学意义(P0.01)。Wnt信号通路抑制剂处理A549细胞后,细胞存活率也下降,细胞凋亡增多,细胞中ROS水平升高,Cleaved Caspase-3水平升高,与正常培养的A549细胞相比差异具有统计学意义(P0.01)。Wnt信号通路抑制剂处理转染PKM2 siRNA后的A549细胞,细胞凋亡率最高,细胞存活率最低,细胞中ROS水平最高,与单纯Wnt信号通路抑制剂处理和单纯转染PKM2 siRNA后的细胞相比,差异具有统计学意义(P0.01)。结论 Wnt信号通路抑制剂和下调PKM2均能够抑制肺癌细胞增殖,促进肺癌细胞凋亡,ROS水平升高,并且二者共同作用后抑制肺癌细胞的作用更明显,下调PKM2可能通过抑制Wnt信号通路阻碍肺癌发生。     

GATA2/GATA6调控胃癌自噬和凋亡的机制

目的 探究GATA2/GATA6调控胃癌自噬和凋亡的机制。方法 以人胃癌SGC-7901细胞为研究对象,建立干扰RNA转染模型,通过流式细胞术检测细胞周期改变,Annexin-V-FITC/PI双染检测细胞凋亡,倒置相差显微镜、透射电镜观察细胞形态,并应用MDC染色对细胞自噬进行测定。结果 GATA2与GATA6表达量与肿瘤直径、淋巴结转移呈正相关,肿瘤直径小于2cm的组织中GATA2与GATA6表达水平显著低于肿瘤直径大于2cm的组织(P0.05)。RT-PCR以及Western-blot实验结果表明,胃癌细胞株GATA2与GATA6的mRNA与蛋白质的表达水平均高于胃黏膜细胞。与人胃黏膜细胞GES1相比,胃癌细胞株中GATA2与GATA6表达水平均显著上调,表明GATA2与GATA6在胃癌中具有较高表达水平。经流式细胞检测发现,对GATA2/GATA6干扰后,胃癌细胞的凋亡水平显著增加(P0.05);CCK-8增殖实验表明GATA2/GATA6敲除后胃癌细胞的增殖能力显著下降(P0.05)。对自噬相关蛋白的检测发现,GATA2/GATA6干扰后胃癌细胞自噬水平显著降低(P0.05),而透射电镜发现,GATA2/GATA6水平降低后,胃癌细胞中自噬小体随之减少。结论 干扰GATA2/GATA6表达可以降低胃癌细胞自噬水平,促进细胞凋亡,从而影响胃癌细胞的增殖能力。     

宫颈、阴道分泌物和血清PKM2检测在子宫内膜癌、宫颈癌诊断中的意义

子宫颈癌和子宫内膜癌是女性生殖系统较为常见的两大恶性肿瘤[1],对患者进行早期诊断、病情监测和治疗具有重要的临床意义[2]。糖类抗原CA125等对子宫颈癌和子宫内膜癌有一定的诊断价值[3,4]。丙酮酸激酶M2型(PKM2)是糖酵解的最后一步限速酶,与肿瘤的发生息息相关[5]。因此,本研究进行宫颈、阴道分泌物和血清PKM2检测分析,为宫颈癌、子宫内膜癌的早期发现、早期诊断提供更为敏感和特异性的检测方法。     

β-微管蛋白及M2型丙酮酸激酶在卵巢浆液性囊腺癌诊断中的意义

微管是组成细胞骨架的主要部分,具有聚合和解聚的动力学特性[1].由α-微管蛋白和β-微管蛋白异二聚体组成[2].γ微管蛋白参与微管的组装,但并非微管的组分.微管的动力学特性使其在细胞有丝分裂起重要作用,其中β-微管蛋白的过表达是恶性肿瘤早期形成的关键[3].Warburg效应指肿瘤细胞在氧气即使充足时亦经糖酵解途径代谢,消耗大量葡萄糖产生乳酸,这种现象被也称为肿瘤的有氧糖酵解[4].M2型丙酮酸激酶（PKM2）即是这一过程的限速酶.它在肿瘤细胞中过表达,并以二聚体形式存在[5].本文通过对比β-微管蛋白及M2型丙酮酸激酶在卵巢浆液性囊腺瘤与卵巢浆液性囊腺癌组织中表达的差别,进一步探索这两种蛋白与卵巢浆液性囊腺癌分期的关系,寻求卵巢浆液性囊腺癌早期诊断新指标.     

Knockdown of UCA1 inhibits viability and glycolysis by suppressing PKM2 expression through the mTOR pathway in non-small cell lung cancer cells

LncRNA urothelial carcinoma associated 1 (UCA1) was reported to be upregulated in non-small cell lung cancer (NSCLC) tissues and contributed to NSCLC progression. Additionally, it has been proposed that the oncogenic role of UCA1 may be related to glucose metabolism in bladder cancer. However, whether and how UCA1 regulates glucose metabolism in the progression of NSCLC remains unknown. Our results showed that knockdown of UCA1 inhibited the viability of NSCLC cells. UCA1 silencing suppressed glycolysis of NSCLC cells by reducing the glucose consumption and lactate production. Additionally, knockdown of UCA1 suppressed PKM2 expression and the mTOR pathway in NSCLC cells. Mechanistically, PKM2 knockdown suppressed the effects of UCA1 on viability and glycolysis of NSCLC cells and inhibition of the mTOR pathway suppressed the effects of UCA1 on viability, glycolysis, and PKM2 expression in NSCLC cells. In conclusion, knockdown of UCA1 inhibited viability and glycolysis by suppressing PKM2 expression maybe through the mTOR pathway in NSCLC cells, providing a novel insight into the molecular mechanism of UCA1 involved in the regulation of glucose metabolism in NSCLC cells.

LncRNA urothelial carcinoma associated 1 (UCA1) was reported to be upregulated in non-small cell lung cancer (NSCLC) tissues and contributed to NSCLC progression.     

Pyruvate kinase M2-specific siRNA induces apoptosis and tumor regression

The development of cancer-specific therapeutics has been limited because most healthy cells and cancer cells depend on common pathways. Pyruvate kinase (PK) exists in M1 (PKM1) and M2 (PKM2) isoforms. PKM2, whose expression in cancer cells results in aerobic glycolysis and is suggested to bestow a selective growth advantage, is a promising target. Because many oncogenes impart a common alteration in cell metabolism, inhibition of the M2 isoform might be of broad applicability. We show that several small interfering (si) RNAs designed to target mismatches between the M2 and M1 isoforms confer specific knockdown of the former, resulting in decreased viability and increased apoptosis in multiple cancer cell lines but less so in normal fibroblasts or endothelial cells. In vivo delivery of siPKM2 additionally causes substantial tumor regression of established xenografts. Our results suggest that the inherent nucleotide-level specificity of siRNA can be harnessed to develop therapeutics that target isoform-specific exons in genes exhibiting differential splicing patterns in various cell types.     

The Utility of [<Superscript>18</Superscript>F]DASA-23 for Molecular Imaging of Prostate Cancer with Positron Emission Tomography

Purpose

There is a strong, unmet need for superior positron emission tomography (PET) imaging agents that are able to measure biochemical processes specific to prostate cancer. Pyruvate kinase M2 (PKM2) catalyzes the concluding step in glycolysis and is a key regulator of tumor growth and metabolism. Elevation of PKM2 expression was detected in Gleason 8–10 tumors compared to Gleason 6–7 carcinomas, indicating that PKM2 may potentially be a marker of aggressive prostate cancer. We have recently reported the development of a PKM2-specific radiopharmaceutical [¹⁸F]DASA-23 and herein describe its evaluation in cell culture and preclinical models of prostate cancer.

Procedure

The cellular uptake of [¹⁸F]DASA-23 was evaluated in a panel of prostate cancer cell lines and compared to that of [¹⁸F]FDG. The specificity of [¹⁸F]DASA-23 to measure PKM2 levels in cell culture was additionally confirmed through the use of PKM2-specific siRNA. PET imaging studies were then completed utilizing subcutaneous prostate cancer xenografts using either PC3 or DU145 cells in mice.

Results

[¹⁸F]DASA-23 uptake values over 60-min incubation period in PC3, LnCAP, and DU145 respectively were 23.4?±?4.5, 18.0?±?2.1, and 53.1?±?4.6 % tracer/mg protein. Transient reduction in PKM2 protein expression with siRNA resulted in a 50.1 % reduction in radiotracer uptake in DU145 cells. Small animal PET imaging revealed 0.86?±?0.13 and 1.6?±?0.2 % ID/g at 30 min post injection of radioactivity in DU145 and PC3 subcutaneous tumor bearing mice respectively.

Conclusion

Herein, we evaluated a F-18-labeled PKM2-specific radiotracer, [¹⁸F]DASA-23, for the molecular imaging of prostate cancer with PET. [¹⁸F]DASA-23 revealed rapid and extensive uptake levels in cellular uptake studies of prostate cancer cells; however, there was only modest tumor uptake when evaluated in mouse subcutaneous tumor models.

     

Evaluation of Glycolytic Response to Multiple Classes of Anti-glioblastoma Drugs by Noninvasive Measurement of Pyruvate Kinase M2 Using [18F]DASA-23

Molecular Imaging and Biology - Pyruvate kinase M2 (PKM2) catalyzes the final step in glycolysis, the key process of tumor metabolism. PKM2 is found in high levels in glioblastoma (GBM) cells with...     

M2型丙酮酸激酶在肿瘤细胞中的多功能性

丙酮酸激酶是葡萄糖代谢的关键限速酶。它主要有4种同工酶形式,L型、R型、M1型和M2型。M2型丙酮酸激酶主要表达于肿瘤细胞当中。肿瘤细胞对葡萄糖的代谢主要是通过有氧糖酵解的形式来完成,这一过程中M2型丙酮酸激酶起着关键限速酶的作用,同时在肿瘤细胞增殖过程中也起着调控信号转导的作用。M2型丙酮酸激酶为肿瘤细胞的增殖提供了能源和物质基础,它可以作为早期检测肿瘤的生物标记物。深入了解M2型丙酮酸激酶在肿瘤中的作用及机制可以为肿瘤靶向治疗提供新的思路。      

TUG1 knockdown enhances adriamycin cytotoxicity by inhibiting glycolysis in adriamycin-resistant acute myeloid leukemia HL60/ADR cells

Taurine-upregulated gene 1 (TUG1) has been reported as an oncogenic long non-coding RNA (lncRNA) in acute myeloid leukemia (AML). Nevertheless, the roles and molecular mechanism of TUG1 in drug resistance of AML cells are still unclear. Glycolysis level was evaluated by detecting glucose consumption and lactate production. qRT-PCR and Western blot were performed to detect TUG1, hexokinase2 (HK2) and pyruvate kinase isoenzyme M2 (PKM2) expressions. Adriamycin (ADR) cytotoxicity and apoptosis were assessed by MTT assay and flow cytometry, respectively. The changes of the protein kinase B (Akt) pathway were determined by Western blot analysis of phosphorylated-Akt (p-Akt) (ser473) and Akt. Our results showed that glycolysis was increased in HL60/ADR cells, as evidenced by the elevated glucose consumption and lactate production, as well as the increased HK2 and PKM2 expressions at mRNA and protein levels. TUG1 was up-regulated in HL60/ADR cells and TUG knockdown inhibited glycolysis. TUG1 knockdown enhanced ADR-induced cytotoxicity and apoptosis in HL60/ADR cells. TUG1 knockdown inhibited the Akt pathway and activation of the Akt pathway by 740Y-P attenuated the effects of TUG1 knockdown on ADR-induced cytotoxicity and apoptosis, as well as glycolysis in HL60/ADR cells. Taken together, TUG1 knockdown enhances adriamycin cytotoxicity in HL60/ADR cells via inhibiting the glycolysis by inactivating the Akt pathway.

Taurine-upregulated gene 1 (TUG1) has been reported as an oncogenic long non-coding RNA (lncRNA) in acute myeloid leukemia (AML).     

Retracted Article: PKM2 overexpression protects against 6-hydroxydopamine-induced cell injury in the PC12 cell model of Parkinson's disease via regulation of the brahma-related gene 1/STAT3 pathway

According to published estimates, pyruvate kinase isoform M2 (PKM2) was expressed in low amounts in patients with Parkinson''s disease (PD). However, the function and molecular mechanism of PKM2 in PD remain largely unknown. The main purpose of our study was to reveal the function and mechanism of PKM2 in the in vitro model of PD. Here, we show that PKM2 decreased in PC12 cells after 6-hydroxydopamine (6-OHDA) treatment, which inhibited PC12 cell survival and induced its apoptosis. PKM2 overexpression is required for 6-OHDA-induced PC12 cell survival. Moreover, up-regulated PKM2 expression suppressed PC12 cell apoptosis and caspase-3 activity compared with the 6-OHDA treatment alone group. Increased brahma-related gene 1 (Brg1) and p-STAT3 expression was observed in PKM2-overexpressed PC12 cells compared to those in 6-OHDA treated PC12 cells. Further studies suggested that Brg1 knockdown impeded the high expression of p-STAT3, which was induced by PKM2 overexpression. Finally, the STAT3 inhibitor reversed the effects of PKM2 on cell survival and apoptosis in 6-OHDA-induced PC12 cells. Our results suggest that PKM2 was involved in 6-OHDA-induced PC12 cell injury by mediating the Brg1/STAT3 pathway.

According to published estimates, pyruvate kinase isoform M2 (PKM2) was expressed in low amounts in patients with Parkinson''s disease (PD) compared with the control health humans.     

Retracted Article: Long noncoding RNA PCA3 regulates glycolysis,viability and apoptosis by mediating the miR-1/CDK4 axis in prostate cancer

Prostate cancer is one of the common tumor malignancies in men worldwide. Although long noncoding RNAs (lncRNAs) have been demonstrated to play essential roles in the progression of prostate cancer, the roles and potential mechanism of lncRNA prostate cancer antigen 3 (PCA3) remain poorly understood. In the present study, we investigated the role of PCA3 in aerobic glycolysis, viability and apoptosis in prostate cancer cells and probed the interaction between PCA3 and microRNA-1 (miR-1)/cyclin-dependent kinase 4 (CDK4). Here we found that PCA3 and CDK4 were up-regulated while miR-1 was down-regulated in prostate cancer tissues and cells. Moreover, knockdown of PCA3 inhibited aerobic glycolysis and viability and induced apoptosis in prostate cancer cells. Intriguingly, PCA3 was bound to miR-1 and inhibition of miR-1 reversed the regulatory effect of PCA3 knockdown on aerobic glycolysis, viability and apoptosis in prostate cancer cells. Besides, CDK4 was indicated as a target of miR-1 and it was regulated by PCA3 through functioning as a competing endogenous RNA (ceRNA) of miR-1 in prostate cancer cells. The results indicated that PCA3 might drive aerobic glycolysis, viability and apoptosis by regulating the miR-1/CDK4 axis in prostate cancer cells, providing a promising avenue for treatment of prostate cancer.

We proved that PCA3 regulated aerobic glycolysis, viability and apoptosis by regulating the miR-1/CDK4 axis in prostate cancer cells.     

肺孤立性结节样腺癌PET/CT表现与丙酮酸激酶M2、硒结合蛋白-1表达的相关性

目的 观察肺孤立性结节样腺癌PET/CT表现与丙酮酸激酶M2(PKM2)、硒结合蛋白-1(SBP-1)表达的相关性。方法 以101例肺孤立性结节样腺癌为观察组,以68例肺良性肿瘤为对照组;比较2组病灶PKM2、SBP-1表达及PET/CT最大标准摄取值(SUV_max)、峰值标准摄取值(SUV_peak)和肿瘤代谢体积(MTV),分析观察组病灶PET/CT表现与PKM2、SBP-1表达的关系。结果 观察组病灶PKM2高表达率高于(P<0.001)、SBP-1高表达率低于对照组(P<0.001);观察组病灶SUV_max、SUV_peak及MTV均高于对照组(P均<0.05)。观察组内PKM2高表达结节呈分叶状、多边形/不规则形、胸膜凹陷征占比及其SUV_max、SUV_peak和MTV均高于PKM2低表达结节(P均<0.05);SBP-1高表达结节呈分叶状、胸膜凹陷征占比及其SUV_max、SUV_peak     

Low-field paramagnetic resonance imaging of tumor oxygenation and glycolytic activity in mice

A priori knowledge of spatial and temporal changes in partial pressure of oxygen (oxygenation; pO(2)) in solid tumors, a key prognostic factor in cancer treatment outcome, could greatly improve treatment planning in radiotherapy and chemotherapy. Pulsed electron paramagnetic resonance imaging (EPRI) provides quantitative 3D maps of tissue pO(2) in living objects. In this study, we implemented an EPRI set-up that could acquire pO(2) maps in almost real time for 2D and in minutes for 3D. We also designed a combined EPRI and MRI system that enabled generation of pO(2) maps with anatomic guidance. Using EPRI and an air/carbogen (95% O(2) plus 5% CO(2)) breathing cycle, we visualized perfusion-limited hypoxia in murine tumors. The relationship between tumor blood perfusion and pO(2) status was examined, and it was found that significant hypoxia existed even in regions that exhibited blood flow. In addition, high levels of lactate were identified even in normoxic tumor regions, suggesting the predominance of aerobic glycolysis in murine tumors. This report presents a rapid, noninvasive method to obtain quantitative maps of pO(2) in tumors, reported with anatomy, with precision. In addition, this method may also be useful for studying the relationship between pO(2) status and tumor-specific phenotypes such as aerobic glycolysis.