肠型胃癌中LDH-A与HDAC1表达的相关性研究

目的 探讨乳酸脱氢酶A（LDH-A）与组蛋白去乙酰化酶1（HDAC1）在肠型胃癌中表达的相关性及其与预后的关系。方法 应用Western blotting法检测HDAC1蛋白在慢病毒介导的LDH A siRNA转染肠型胃癌细胞株SGC7901中表达的变化;应用免疫组织化学方法检测661例肠型胃癌组织及癌旁正常组织中LDH-A与HDAC1蛋白的表达情况并分析其表达与预后的关系。结果 Western blotting检测结果显示,LDH-A蛋白在SGC7901细胞株中的表达明显上调,且LDH-A的沉默可显著下调HDAC1的表达。肠型胃癌组织中LDH-A蛋白的高表达率为54.8％（362／661）,明显高于癌旁正常组织的12.9％（85／661）,两者差异有统计学意义（P＜0.01）;肠型胃癌组织中HDAC1的高表达率为51.3％（339／661）,明显高于癌旁正常组织的15.4％（102／661）,两者差异有统计学意义（P＜0.01）。LDH-A与HDAC1蛋白在肠型胃癌组织中的表达呈正相关（r=0.324, P＜0.001）。单因素生存分析结果显示,LDH-A与HDAC1均低表达患者的生存曲线明显优于其他组合（P＜0.001）;多因素生存分析结果显示LDH-A与HDAC1表达均为肠型胃癌独立的预后因素。结论在肠型胃癌中, LDH-A与HDAC1的表达呈正相关,采用LDH-A和HDAC1双靶点抑制治疗可能具有潜在的生存获益。     

LDH-A negatively regulates dMMR in colorectal cancer

Although immune checkpoint inhibitors (ICIs) have achieved unprecedented success in dMMR tumors, pMMR tumors accounting for 85% of colorectal cancer (CRC) cases remain unresponsive. Lactate dehydrogenase A (LDH-A) is the rate-limiting enzyme that catalyzes the transformation of pyruvate to lactate in the process of glycolysis. We investigated the relationship between LDH-A and dMMR with the purpose of exploring the treatment strategy for pMMR CRC patients. We here show that LDH-A can promote the proliferation of dMMR and pMMR CRC cells by positively regulating MMR proteins both in vitro and in vivo. LDH-A inhibition can improve the efficacy of PD-1 blockade in a pMMR CRC xenograft model. A statistical analysis of 186 CRC specimens showed a significant correlation between LDH-A and dMMR status. Moreover, patients with both low LDH-A expression and dMMR exhibited better disease-free survival compared with patients with other combinations. The close correlation of LDH-A and dMMR may offer a promising therapeutic strategy in which the combination of LDH-A inhibitor and ICIs may improve the clinical benefit for pMMR CRC patients.     

Nanog、Sox2、TFF3在肠型胃癌中的表达及其与肠型胃癌预后关系的研究

目的 探讨Nanog、Sox2、TFF3在肠型胃癌中的表达及其与肠型胃癌预后的关系.方法 回顾性分析60例肠型胃癌患者的临床资料,应用免疫组化法检测Nanog、Sox2、TFF3在肠型胃癌患者中的表达情况,分析其表达情况与患者临床病理特征及预后的关系.结果60例肠型胃癌患者中,Nanog高表达者38例(63.3%)、TFF3高表达者33例(55.0%),Sox2低表达者35例(58.3%).单因素分析结果显示,Nanog、TFF3、Sox2的表达情况与肠型胃癌患者的肿瘤浸润深度、淋巴结转移情况及TNM分期有关(P﹤0.05),与肠型胃癌患者的年龄、性别、肿瘤部位无关(P﹥0.05).多因素分析结果显示,Nanog、Sox2、TFF3的表达情况与肠型胃癌患者的无病生存期和总生存期有关,同时具有Nanog高表达、TFF3高表达、Sox2低表达的肠型胃癌患者预后最差.结论Nanog、Sox2、TFF3可成为评价肠型胃癌患者预后的生物标志物,联合检测更利于判断肠型胃癌患者的预后.     

八聚体转录因子4蛋白表达与非肌层浸润性膀胱癌的关系

目的 探讨八聚体转录因子4(Oct4)蛋白在非肌层浸润性膀胱癌中的表达情况,并分析其表达与患者临床病理特征及预后的关系.方法 采用免疫组化生物素酶标法检测87例非肌层浸润性膀胱移行细胞癌和15例癌旁正常组织中Oct4蛋白的表达情况,并结合临床资料分析Oct4蛋白的表达在膀胱癌患者中的意义.结果 正常组织中Oct4蛋白的表达水平明显低于肿瘤组织(P＜0.01).G1级、G2级、G3级膀胱癌的Oct4蛋白阳性表达率分别为40.7％、69.4％、91.7％,差异有统计学意义(P＜0.01).所有患者随访3～78个月,其中63例出现复发,复发组与未复发组中Oct4蛋白的阳性表达率分别为77.8％和37.5％,差异有统计学意义(P＜0.0l);21例复发患者出现进展,进展组与未进展组中Oct4蛋白的阳性表达率分别为71.4％和65.2％,差异无统计学意义(P＞0.05).Oct4蛋白的表达与患者的年龄、性别、肿瘤的数目及大小无明显相关性(P＞0.05).结论 Oct4蛋白检测有助于早期发现膀胱肿瘤,并可用来判断肿瘤的恶性分化程度及检测患者术后的复发情况.     

Oct4蛋白在左右半结肠癌中表达的比较及预后分析

  目的  探讨人结肠癌组织中Oct4蛋白表达情况,并分析其预后价值。  方法  选取2008年1月至2011年12月于无锡市锡山人民医院行左半结肠癌根治术的标本89例,右半结肠癌根治术的标本77例,共166例。应用免疫组织化学检测全部结肠癌组织中Oct4蛋白的表达,并对病理参数及预后情况进行分析。  结果  免疫组织化学法染色结果显示Oct4蛋白在左半结肠癌组织中阳性表达率为68.54%,右半结肠癌组织中阳性表达率为71.43%,两者无显著差异（P>0.05）;Oct4表达水平在右半结肠癌组织中与组织学分级、淋巴结转移及Dukes分期呈正相关,而在左半结肠癌组织中仅与组织学分级及Dukes分期呈正相关,与淋巴结转移情况无显著相关;另外,左半结肠癌患者5年生存率明显高于右半结肠癌患者（P < 0.01）,且Oct4表达与5年总体生存情况无显著相关;右半结肠癌患者中,Oct4过表达组5年总体生存明显低于阴性组（P < 0.05）。Cox生存分析提示仅右半结肠癌中高水平Oct4提示着预后不佳。  结论  Oct4在左、右半结肠癌进展中起到了不同的作用,为探索结肠癌诊疗新靶点提供理论依据。      

八聚体转录因子4蛋白表达与非肌层浸润性膀胱癌的关系

目的 探讨八聚体转录因子4(Oct4)蛋白在非肌层浸润性膀胱癌中的表达情况,并分析其表达与患者临床病理特征及预后的关系.方法 采用免疫组化生物素酶标法检测87例非肌层浸润性膀胱移行细胞癌和15例癌旁正常组织中Oct4蛋白的表达情况,并结合临床资料分析Oct4蛋白的表达在膀胱癌患者中的意义.结果 正常组织中Oct4蛋白的...     

Oct4与EGFR在右半结肠癌组织中表达的相关性及临床价值

目的:探讨Oct4与EGFR在右半结肠癌组织中表达的相关性及临床价值.方法:应用免疫组织化学法检测120例右半结肠癌组织中Oct4与EGFR的表达,并与病理参数进行相关性分析;结合GEPIA数据库结果确认两者相关性.结果:免疫组化染色结果显示,Oct4表达与组织学分级、淋巴结转移以及TNM分期呈正相关(P＜0.05),...     

LDH-A silencing suppresses breast cancer tumorigenicity through induction of oxidative stress mediated mitochondrial pathway apoptosis

LDH-A, as the critical enzyme accounting for the transformation from pyruvate into lactate, has been demonstrated to be highly expressed in various cancer cells and its silencing has also been approved relating to increased apoptosis in lymphoma cells. In this study, we intend to investigate the correlation between LDH-A and other clinicopathological factors of breast cancer and whether LDH-A silencing could suppress breast cancer growth, and if so the potential mechanisms. 46 breast cancer specimens were collected to study the relation between LDH-A expression and clinicopathological characteristics including menopause, tumor size, node involvement, differentiation, and pathological subtypes classified by ER, PR, and Her-2. shRNAs were designed and applied to silence LDH-A expression in breast cancer cell lines MCF-7 and MDA-MB-231. The effects of LDH-A reduction on cancer cells were studied by a series of in vitro and in vivo experiments, including cell growth assay, apoptosis evaluation, oxidative stress detection, transmission electron microscopy observation, and tumor formation assay on nude mice. LDH-A expression was found to correlate significantly with tumor size and to be independent for other clinicopathological factors. LDH-A reduction resulted in an inhibited cancer cell proliferation, elevated intracellular oxidative stress, and induction of mitochondrial pathway apoptosis. Meanwhile, the tumorigenic ability of LDH-A deficient cancer cells was significantly limited in both breast cancer xenografts. The Ki67 positive cancer cells were significantly reduced in LDH-A deficiency tumor samples, while the apoptosis ratio was enhanced. Our results suggested that LDH-A inhibition might offer a promising therapeutic strategy for breast cancer.     

Inhibition of the Warburg effect with a natural compound reveals a novel measurement for determining the metastatic potential of breast cancers

Metabolism is an important differentiating feature of cancer cells. Lactate dehydrogenases (LDH) A/B are metabolically important proteins and are involved in the critical step of inter-conversion of lactate to pyruvate. Panepoxydone (PP), a natural NF-kB inhibitor, significantly reduces the oxygen consumption and lactate production of MCF-7 and triple negative (MDA-MB-231, MDA-MB-468 and MDA-MB-453) breast cancer cells. We further observed that PP inhibited mitochondrial membrane potential and the ATP synthesis using flow cytometry. PP also up-regulated LDH-B and down-regulated LDH-A expression levels in all breast cancer cells to similar levels observed in HMEC cells. Over-expression of LDH-B in cancer cell lines leads to enhanced apoptosis, mitochondrial damage, and reduced cell migration. Analyzing the patient data set GDS4069 available on the GEO website, we observed 100% of non TNBC and 60% of TNBC patients had less LDH-B expression than LDH-A expression levels. Herein we report a new term called Glycolytic index, a novel method to calculate utilization of oxidative phosphorylation in breast cancer cells through measuring the ratio of the LDH-B to LDH-A. Furthermore, inhibitors of NF-kB could serve as a therapeutic agent for targeting metabolism and for the treatment of triple negative breast cancer.     

Increased Expression of Oct4, Nanog and CD24 Predicts Poor Response to Chemo-Radiotherapy and Unfavourable Prognosis in Locally Advanced Oral Squamous Cell Carcinoma

Background: Current study investigates the role of Oct4, Nanog and CD24 in locally-advanced oral squamous cell carcinoma (OSCC), to evaluate whether the expression of these markers can predict efficacy of neoadjuvant-chemo-radiotherapy and survival of patients. Methods: Biomarker expression was evaluated in 50 homogenously treated patients of locally-advanced OSCC. Results: Clinical response was complete in 30% (n=15), partial response in 46% (n=23), no response in 24% (n=12). Pathologically, 74% patents (n=37) were responders and 26% were non-responders (n=13). Biomarker-overexpression was seen in 46% cases for Oct4, 54% cases for Nanog and 58% cases for CD24. Oct4, Nanog and CD24 expression showed significant correlation with clinical and pathological response (p <0.05). Three year recurrence-free survival was 71%, overall survival was 66%. Post-treatment advanced pathological N (ypN), post treatment advanced pathological TNM (ypTNM) stage, clinical non-response, pathologic non-response, positive/high expression of all three biomarkers had a significant negative impact on recurrence-free and overall survival. Conclusions: Expressions of Oct4, Nanog and CD24 have significant association with treatment response and survival in patients with locally advanced OSCC treated with neoadjuvant chemo-radiation. Survival of these patients is significantly affected by ypN stage, ypTNM stage, expression of all three biomarkers, clinical and pathological response to neoadjuvant therapy.     

Comparative Analysis of Oct4 in Different Histological Subtypes of Esophageal Squamous Cell Carcinomas in Different Clinical Conditions

Background: Esophageal squamous cell carcinoma (ESCC) is a common cancer with poor prognosis. It hasbeen hypothesized that Oct4 positive radioresistant stem cells may be responsible for tumor recurrence. Hence,we evaluated Oct4 expression in ESCC in pre-treatment, post neo-adjuvant residual and post-surgical recurrenttumours. Materials and Methods: Endoscopic mucosal biopsies were used to study Oct4 expression and theobservations were correlated with histological tumor grades, patient data and clinical background. Results: Allpatients presented with dysphagia with male predominance and a wide age range. Majority of the patients hadintake of mixed diet, history of alcohol and tobacco intake was documented in less than half of the patients. Oct4 expression was significantly higher in poorly differentiated (PDSCC) and basaloid (BSCC) subtypes than theother better differentiated tumor morphology. Oct4 was also expressed by adjoining esophageal mucosa showinglow grade dysplasia and basal cell hyperplasia (BCH). Biopsies in PDSCC and BSCC groups were more likelyto show a positive band for Oct4 by polymerase chain reaction (PCR). Dysplasia and BCH mucosa also showedOct4 positivity by PCR. All mucosal biopsies with normal morphology were negative for Oct4. Number of tissuesamples showing Oct4 positivity by PCR was higher than that by the conventional immunohistochemistry (p>0.05).Oct4 expression pattern correlated only with tumor grading, not with other parameters including the clinicalbackground or patient data. Conclusions: Our observations highlighted a possible role of Oct4 in identifyingputative cancer stem cells in ESCC pathobiology and response to treatment. The implications are either in vivoexistence of Oct4 positive putative cancer stem cells in ESCC or acquisition of cancer stem cell properties bytumor cells as a response to treatment given, resulting ultimately an uncontrolled cell proliferation and treatmentfailure.     

On-target Inhibition of Tumor Fermentative Glycolysis as Visualized by Hyperpolarized Pyruvate

Many cancer cells display the Warburg effect, that is, enhanced glycolysis followed by fermentation (conversion of pyruvate to lactate). Recently, the molecular basis for these effects has started to be elucidated, and the up-regulation of the lactate dehydrogenase A (LDH-A) isoform of lactate dehydrogenase is felt to be a major molecular mediator of this phenomenon. Moreover, LDH-A expression in tumor tissue and LDH-A levels in blood portend a bad prognosis, and LDH-A blockade can lead to tumor growth inhibition in tumor transplant models. We have extended existing data (some of which were published during the time when we were carrying out our studies) in two important ways: 1) inhibition of LDH-A in a glycolytic lung cancer cell line results in reactive oxygen species-mediated apoptosis and increased sensitivity to the chemotherapeutic drug paclitaxel and 2) inhibition of fermentative glycolysis can also be accomplished by activation of the pyruvate dehydrogenase complex by the drug dichloroacetate, now undergoing clinical trials, and that this phenomenon can be monitored in vivo in a noninvasive real-time manner through magnetic resonance spectroscopy using hyperpolarized pyruvate. Collectively, these data suggest that in vivo effects of drugs that redirect the fate of pyruvate, and hence are aimed at reversing the Warburg effect, could be monitored through the use of hyperpolarized magnetic resonance spectroscopy, a method that is scalable to human use.     

肝癌细胞系中Oct4与Wnt/β-catenin 和TGF-β信号通路的相互影响

 目的 探讨干细胞相关基因Oct4与Wnt/β-catenin 及TGF-β信号通路在肝癌细胞系中的相互作用。方法 应用RT PCR法检测Oct4、Wnt/β-catenin及TGF-β信号通路相关基因β-catenin、Wnt10b、TCF3及 ELF、Smad3和 Smad4在肝癌组织及细胞系中的表达;使用siRNA 沉默人肝癌HepG2细胞Oct4和TCF3的表达,实时荧光定量RT PCR法检测Wnt10b、β-catenin、 TCF3及 ELF、Smad3和 Smad4等基因的表达变化。结果 Oct4和β catenin、Wnt10b、TCF3及 ELF、Smad3和 Smad4在肝癌组织及细胞系中同时表达; siRNA-Oct4 沉默人肝癌HepG2细胞Oct4后,Oct4表达明显下调,β-catenin、Wnt10b随之下调达40%~50%左右,而TCF3表达升高到3倍左右;同时ELF、Smad3和 Smad4均下降到原来的1%以下;而siRNA-TCF3 沉默TCF3后,Oct4的表达也升高2~3倍;ELF升高亦达2~3倍,Smad3和 Smad4也略有升高。结论 Oct4和β-catenin、Wnt10b、TCF3及 ELF、Smad3和 Smad4在肝癌组织及细胞系中同时表达提示彼此之间有相互作用。RNAi实验证明Oct4对Wnt/β-catenin及TGF-β信号通路的成员有调控作用;Oct4 与TCF3之间的负反馈作用值得深入研究。     

转录因子Oct4、Sox2在结肠癌中的表达及意义

目的：研究转录因子Sox2、Oct4在结肠癌中的表达及相互关系,探索两者参与结肠癌发生发展中的临床意义。方法：用免疫组织化学SP法检测Oct4、Sox2在50例结肠癌及其匹配的癌旁组织中的表达。West-erRblot检测其在结肠癌细胞株SW480、SW620、Lovo及永生化结肠上皮细胞株HIEC中的表达。结果：免疫组织化学结果显示Oct4、Sox2在结肠癌组织中阳性表达率分别为80％（40／50）,74％（37／50）,显著高于匹配的癌旁组织,后者表达率分别为48％（24／50）,20％（10／50）（P〈0．05）,并且两者阳性表达率呈正相关关系（r=0．465,P〈0．05）。Oct4表达与病理级别呈负相关,而与年龄、性别无相关性。Sox2表达与患者的年龄、性别、病理级别无明显相关性。Westernblot显示结肠癌细胞株与永生化结肠上皮细胞HIEC均表达Oct4,Sox2;0et4、Sox2在结肠癌高转移细胞系SW620较亲本细胞SW480表达增高。结论：转录因子Oct4、Sox2在结肠癌组织中表达高于癌旁组织,并且两者的表达存在正相关性。提示Oct4和Sox2在结肠癌的发生中起着重要的作用。     

人脑胶质瘤组织中Oct4、Wnt2的表达及其临床意义

目的: 探讨Oct4和Wnt2在人脑胶质瘤组织中的表达及其与临床病理特征的关系。方法: 选取临床及病理资料完整的长江大学附属第一医院2006-2009年手术切除并经病理证实为胶质瘤的石蜡标本56例,采用免疫组织化学方法检测56例胶质瘤组织（其中15例为术后复发）和10例脑外伤行内减压术切除的脑组织标本Oct4、Wnt2的表达。结果: 正常脑组织中Oct4表达均为阴性,Wnt2表达仅1例呈弱阳性;56例胶质瘤组织中Oct4阳性34例（60.7%）,Wnt2阳性40例（714%）。Oct4、Wnt2在低度恶性胶质瘤组中和高度恶性胶质瘤中的阳性率分别为46.2%和73.3%（P<0.05）及577%和83.3%（P<0.05）,Oct4在复发肿瘤和初诊肿瘤中的阳性率分别为86.7%和51.2%（P<0.05）。人脑胶质瘤组织中 Oct4和Wnt2表达呈正相关（r=0.537,P<0.01）。结论: Oct4、Wnt2的表达与人脑胶质瘤的恶性程度相关,Oct4的表达与胶质瘤的复发相关;Oct4可能是通过Wnt通路而参与了胶质瘤的发生、发展。     

CIP2A is an Oct4 target gene involved in head and neck squamous cell cancer oncogenicity and radioresistance

Radiotherapy is a mainstay for treatment of many human cancer types, including head and neck squamous cell carcinoma (HNSCC). Thereby, it is clinically very relevant to understand the mechanisms determining radioresistance. Here, we identify CIP2A as an Oct4 target gene and provide evidence that they co-operate in radioresistance. Oct4 positively regulates CIP2A expression both in testicular cancer cell lines as well as in embryonic stem cells. To expand the relevance of these findings we show that Oct4 and CIP2A are co-expressed in CD24 positive side-population of patient-derived HNSCC cell lines. Most importantly, all Oct4 positive HNSCC patient samples were CIP2A positive and this double positivity was linked to poor differentiation level, and predicted for decreased patient survival among radiotherapy treated HNSCC patients. Oct4 and CIP2A expression was also linked with increased aggressiveness and radioresistancy in HNSCC cell lines. Together we demonstrate that CIP2A is a novel Oct4 target gene in stem cells and in human cancer cell lines. Clinically these results suggest that diagnostic evaluation of HNSCC tumors for Oct4 or Oct4/CIP2A positivity might help to predict HNSCC tumor radioresistancy. These results also identify both Oct4 and CIP2A as potential targets for radiosensitation.     

The Warburg effect as a therapeutic target for bladder cancers and intratumoral heterogeneity in associated molecular targets

Bladder cancer is the 10th most common cancer worldwide. For muscle-invasive bladder cancer (MIBC), treatment includes radical cystectomy, radiotherapy, and chemotherapy; however, the outcome is generally poor. For non–muscle-invasive bladder cancer (NMIBC), tumor recurrence is common. There is an urgent need for more effective and less harmful therapeutic approaches. Here, bladder cancer cell metabolic reprogramming to rely on aerobic glycolysis (the Warburg effect) and expression of associated molecular therapeutic targets by bladder cancer cells of different stages and grades, and in freshly resected clinical tissue, is investigated. Importantly, analyses indicate that the Warburg effect is a feature of both NMIBCs and MIBCs. In two in vitro inducible epithelial-mesenchymal transition (EMT) bladder cancer models, EMT stimulation correlated with increased lactate production, the end product of aerobic glycolysis. Protein levels of lactate dehydrogenase A (LDH-A), which promotes pyruvate enzymatic reduction to lactate, were higher in most bladder cancer cell lines (compared with LDH-B, which catalyzes the reverse reaction), but the levels did not closely correlate with aerobic glycolysis rates. Although LDH-A is expressed in normal urothelial cells, LDH-A knockdown by RNAi selectively induced urothelial cancer cell apoptotic death, whereas normal cells were unaffected—identifying LDH-A as a cancer-selective therapeutic target for bladder cancers. LDH-A and other potential therapeutic targets (MCT4 and GLUT1) were expressed in patient clinical specimens; however, positive staining varied in different areas of sections and with distance from a blood vessel. This intratumoral heterogeneity has important therapeutic implications and indicates the possibility of tumor cell metabolic coupling.