原发性口腔鳞癌组织及血清中内皮抑素表达的意义

目的:探讨原发性口腔鳞癌患者组织和血清中内皮抑素表达及与肿瘤分期、分级的关系。方法:采用免疫组化方法检测36例口腔鳞癌和12例正常口腔粘膜组织中内皮抑素表达情况。ELISA法检测36例口腔鳞癌患者术前血清内皮抑素水平,14例健康者血清做对照。结果:内皮抑素主要见于肿瘤组织细胞质。正常口腔粘膜中内皮抑素表达率为7.15%,口腔鳞癌组织中内皮抑素阳性率为76.44%,其中G1、G2、G3级阳性率分别为47.21%、79.17%、90.90%,病理分级间比较差异有统计学意义(P<0.05)。口腔鳞癌患者血清中内皮抑素水平(49.62±1.72)ng/mL显著高于健康对照者(5.60±0.37)ng/mL(P<0.05),TNM分期III、IV期肿瘤患者血清内皮抑素水平显著高于I和II期(P<0.05)。结论:口腔鳞癌患者组织和血清中内皮抑素表达显著升高,并与肿瘤分期、分级相关,检测内皮抑素表达有助于判断口腔鳞癌恶性程度。     

原发性口腔鳞癌组织及血清中内皮抑素表达的意义

目的：探讨原发性口腔鳞癌患者组织和血清中内皮抑素表达及与肿瘤分期、分级的关系。方法：采用免疫组化方法检测36例口腔鳞癌和12例正常口腔粘膜组织中内皮抑素表达情况。ELISA法检测36例口腔鳞癌患者术前血清内皮抑素水平,14例健康者血清做对照。结果：内皮抑素主要见于肿瘤组织细胞质。正常口腔粘膜中内皮抑素表达率为7.15%,口腔鳞癌组织中内皮抑素阳性率为76.44%,其中G1、G2、G3级阳性率分别为47.21%、79.17%、90.90%,病理分级间比较差异有统计学意义（P〈0.05）。口腔鳞癌患者血清中内皮抑素水平（49.62±1.72）ng/mL显著高于健康对照者（5.60±0.37）ng/mL（P〈0.05）,TNM分期III、IV期肿瘤患者血清内皮抑素水平显著高于I和II期（P〈0.05）。结论：口腔鳞癌患者组织和血清中内皮抑素表达显著升高,并与肿瘤分期、分级相关,检测内皮抑素表达有助于判断口腔鳞癌恶性程度。     

VEGF-C 在口腔鳞状细胞癌中的表达及其意义

目的:检测血管内皮生长因子-C(VEGF-C)在口腔鳞状细胞癌(鳞癌)和癌旁组织中的表达情况,探讨VEGF-C在口腔鳞癌的增殖、浸润和淋巴转移中的作用。方法:采用免疫组化方法检测60例口腔鳞癌病人癌组织和癌旁组织中VEGF-C的表达,应用图像分析系统进行分析,用Spearman相关分析研究其与病变部位、肿瘤大小、病理分级、临床分期及颈淋巴结转移之间的关系。结果:口腔鳞癌组织VEGF-C的表达明显高于癌旁组织(u=7.747,P<0.01),其表达强度与临床分期及淋巴结转移密切相关(r=0.564、0.706,P<0.05),与病变部位、大小、病理分级无关。结论:口腔鳞癌细胞分泌VEGF-C诱导癌周淋巴管增生扩张是发生区域淋巴结转移的重要因素之一,VEGF-C有望作为早期临床判断和预测颈淋巴结转移的指标之一。     

Anxa2, Stat3 在口腔鳞状细胞癌中的表达及意义

目的:探讨膜联蛋白A2(Anxa2)和细胞信号转导和转录激活因子3(Stat3)在口腔鳞癌组织中的表达及其临床意义。方法:选择口腔鳞癌石蜡标本80例为实验组,20例正常口腔黏膜组织为对照组。应用免疫组织化学法检测Anxa2和Stat3蛋白的阳性表达,并进行结果判定,采用x2和Spearman等级相关分析法分析二者表达差异性及其相关性。结果:Anxa2、Stat3在病例组中的阳性表达率,分别为81.3%(65/80)、87.5%(70/80),明显高于对照组中的表达率,25.0%(5/20)、30.0%(6/20),且差异有统计学意义(P0.05),Anxa2和Stat3蛋白表达水平与口腔鳞癌的分期、淋巴结转移及分化程度有密切关系。Spearman等级相关分析Anxa2和Stat3在口腔鳞癌组织中蛋白的表达呈正相关(r=0.302,P0.01)。结论:Anxa2、Stat3在OSCC的发生和转移过程中均具有重要的作用,且二者间也有相互作用的关系。     

CD9在口腔鳞状细胞癌中的表达水平及其临床意义

目的:口腔鳞状细胞癌是一类极易发生局部侵袭和淋巴结转移的恶性肿瘤,CD9蛋白在多种肿瘤的发生发展及侵袭转移过程中起到重要作用,本研究旨在分析CD9蛋白在口腔鳞状细胞癌中的表达水平及其临床意义。方法:收集我院诊断明确的口腔鳞癌肿瘤患者石蜡标本合计80例,通过免疫组化手段对CD9蛋白表达水平进行评价,并根据CD9蛋白的表达水平分组,分析患者的临床病理学特征与CD9蛋白的关系。结果:CD9在正常组织和癌旁组织正常表达,在肿瘤组织中表达率低,其表达水平和口腔鳞癌的分化程度,淋巴结转移及最终分期有相关性(P0.05)。结论:本研究结果揭示,CD9在口腔鳞状细胞癌的发生发展中起到重要作用,CD9蛋白水平的低表达或不表达可能预测着肿瘤具有更明显的恶性生物学行为,并可能成为口腔鳞状细胞癌预后的生物学指标及基因治疗的新靶点。     

VEGF在口腔鳞状细胞癌中的表达及其临床意义

目的:研究血管内皮生长因子(Vascular endothelial growth factor,VEGF)在口腔鳞状细胞癌中的表达及其临床意义.方法:免疫组化EnVisionTM法检测VEGF在57例手术治疗的原发性OSCC(口腔鳞状细胞癌)中的表迭.结果:①VEGF在OSCC中的阳性表达率为40.35%.②VEGF在不同发病部位组的表达:牙龈癌>舌癌>颊癌>唇癌,组间无统计学差异(P>0.05).③VEOF在不同病理分级组、不同预后组、不同年龄组、不同性别组的表达无显著性差异(P>0.05);淋巴结转移组的表达显著高于无转移组(P<0.05).结论:虽然VEGF的高表达可能在淋巴结转移中起一定的作用,但还不能作为预测口腔鳞状细胞癌临床生物学行为及预后的可靠指标.     

High PD-L1 Expression Correlates with Metastasis and Poor Prognosis in Oral Squamous Cell Carcinoma

PD-L1 has been widely demonstrated to contribute to failed antitumor immunity. Blockade of PD-L1 with monoclonal antibody could modulate the tumor immune environment to augment immunotherapy. PD-L1 expression is also detected in several types of cancer and is associated with poor prognosis. However, the prognostic role of PD-L1 in oral squamous cell carcinoma (OSCC) is still controversial. Our aim was to determine the role of PD-L1 in the prognosis of OSCC patients to identify its potential therapeutic relevance. PD-L1 immunoreactivity was analyzed by immunohistochemistry in 305 cancer specimens from primary OSCC patients. The medium follow-up time after surgery was 3.8 years (range from 0.1 to 11.1 years). The prognostic value of PD-L1 on overall survival was determined by Kaplan-Meier analysis and Cox proportional hazard models. Higher PD-L1 expression is more likely in tumor tissues of female than male OSCC patients (P = 0.0062). Patients with distant metastasis also had high PD-L1 expression (P = 0.0103). Multivariate analysis identified high PD-L1 expression as an independent risk factor in males and smokers (males: hazard ratio = 1.556, P = 0.0077; smokers: hazard ratio = 2.058, P = 0.0004). We suggest that PD-L1 expression, determined by IHC staining, could be an independent prognostic marker for OSCC patients who are male or who have a smoking habit.     

Antitumor Effect of TRAIL on Oral Squamous Cell Carcinoma using Magnetic Nanoparticle-Mediated Gene Expression

We developed a new magnetic nanovector to improve the efficiency and targeting of transgene therapy for oral squamous cell carcinoma (OSCC). Positively charged polymer PEI-modified Fe₃O₄ magnetic nanoparticles were tested as gene transfer vectors in the presence of a magnetic field. The Fe₃O₄ nanoparticles were prepared by a co-precipitation method and had good dispersibility in water. These nanoparticles modified by PEI were combined with negatively charged pACTERT-EGFP via electrostatic interaction. The transfection efficiency of the magnetic nano-gene vector with the magnetic field was determined by a fluorescence-inverted microscope and flow cytometry. The results showed significant improvement compared with the control group (p < 0.05). The magnetic complexes also exhibited up to 6-times higher transfection efficiency compared with commonly used PEI or lipofectin. On the basis of these results, the antitumor effect with suicide gene therapy using pACTERT-TRAIL in vitro and vivo was evaluated. In vitro apoptosis was determined with the Annexin V-FITC Apoptosis Detection Kit. The results suggested that PEI-modified Fe₃O₄ nanoparticles could mediate the killing of Tca83 cells. Furthermore, treatment with pACTERT-TRAIL delivered by magnetic nanoparticles showed a significant cytostatic effect through the induction of apoptosis in a xenograft model. This indicates that magnetic nano-gene vectors could improve the transgene efficiency for Tca83 cells and could exhibit antitumor functions with the plasmid pACTERT-TRAIL. This may be a new way to treat OSCC.     

The Nucleolar Protein Myb-binding Protein 1A (MYBBP1A) Enhances p53 Tetramerization and Acetylation in Response to Nucleolar Disruption

Tetramerization of p53 is crucial to exert its biological activity, and nucleolar disruption is sufficient to activate p53. We previously demonstrated that nucleolar stress induces translocation of the nucleolar protein MYBBP1A from the nucleolus to the nucleoplasm and enhances p53 activity. However, whether and how MYBBP1A regulates p53 tetramerization in response to nucleolar stress remain unclear. In this study, we demonstrated that MYBBP1A enhances p53 tetramerization, followed by acetylation under nucleolar stress. We found that MYBBP1A has two regions that directly bind to lysine residues of the p53 C-terminal regulatory domain. MYBBP1A formed a self-assembled complex that provided a molecular platform for p53 tetramerization and enhanced p300-mediated acetylation of the p53 tetramer. Moreover, our results show that MYBBP1A functions to enhance p53 tetramerization that is necessary for p53 activation, followed by cell death with actinomycin D treatment. Thus, we suggest that MYBBP1A plays a pivotal role in the cellular stress response.     

Oncogenic MicroRNA-155 Down-regulates Tumor Suppressor CDC73 and Promotes Oral Squamous Cell Carcinoma Cell Proliferation: IMPLICATIONS FOR CANCER THERAPEUTICS*

The CDC73 gene is mutationally inactivated in hereditary and sporadic parathyroid tumors. It negatively regulates β-catenin, cyclin D1, and c-MYC. Down-regulation of CDC73 has been reported in breast, renal, and gastric carcinomas. However, the reports regarding the role of CDC73 in oral squamous cell carcinoma (OSCC) are lacking. In this study we show that CDC73 is down-regulated in a majority of OSCC samples. We further show that oncogenic microRNA-155 (miR-155) negatively regulates CDC73 expression. Our experiments show that the dramatic up-regulation of miR-155 is an exclusive mechanism for down-regulation of CDC73 in a panel of human cell lines and a subset of OSCC patient samples in the absence of loss of heterozygosity, mutations, and promoter methylation. Ectopic expression of miR-155 in HEK293 cells dramatically reduced CDC73 levels, enhanced cell viability, and decreased apoptosis. Conversely, the delivery of a miR-155 antagonist (antagomir-155) to KB cells overexpressing miR-155 resulted in increased CDC73 levels, decreased cell viability, increased apoptosis, and marked regression of xenografts in nude mice. Cotransfection of miR-155 with CDC73 in HEK293 cells abrogated its pro-oncogenic effect. Reduced cell proliferation and increased apoptosis of KB cells were dependent on the presence or absence of the 3′-UTR in CDC73. In summary, knockdown of CDC73 expression due to overexpression of miR-155 not only adds a novelty to the list of mechanisms responsible for its down-regulation in different tumors, but the restoration of CDC73 levels by the use of antagomir-155 may also have an important role in therapeutic intervention of cancers, including OSCC.     

Oral Squamous Cell Carcinoma in a Pigtailed Macaque (Macaca nemestrina)

An adult, gravid, female pigtailed macaque (Macaca nemestrina) presented for facial swelling centered on the left mandible that was approximately 5 cm wide. Differential diagnoses included infectious, inflammatory, and neoplastic origins. Definitive antemortem diagnosis was not possible, and the macaque''s condition worsened despite supportive care. Necropsy findings included a mandibular mass that was locally invasive and expansile, encompassing approximately 80% of the left mandibular bone. The mass replaced portions of the soft palate, hard palate, sinuses, ear canal, and the caudal–rostral calvarium and masseter muscle. Histologically, the mass was a neoplasm that was poorly circumscribed, unencapsulated, and infiltrative invading regional bone and soft tissue. The mass consisted of polygonal squamous epithelial cells with intercellular bridging that breached the epithelial basement membrane and formed invasive nests, cords, and trabeculae. The mitotic rate averaged 3 per 400× field of view, with occasional bizarre mitotic figures. Epithelial cells often exhibited dyskeratosis, and the nests often contained compact lamellated keratin (keratin pearls). The neoplasm was positive via immunohistochemistry for pancytokeratin, variably positive for S100, and negative for vimentin, smooth muscle actin, and desmin. The gross, histologic, and immunohistochemical findings were consistent with an aggressive oral squamous cell carcinoma. The neoplasm was negative via PCR for papilloma virus. In general, neoplasia in macaques is rare. Although squamous cell carcinomas are one of the most common oral neoplasia in many species, to our knowledge this case represents the first reported oral squamous cell carcinoma in a pigtailed macaque.Abbreviation: SCC, squamous cell carcinomaSquamous cell carcinomas (SCC) are one of the most commonly reported oral tumors. They are characterized as firm, nodular to irregular, soft-tissue masses that are often ulcerated.⁶ These tumors are frequently highly invasive to local bone and muscle and occasionally metastasize to local and regional lymph nodes.⁶ Histologically, SCC are characterized by keratin pearls, intercellular bridges, and positive cytokeratin staining on immunohistochemistry.^6,18 SCC have been associated with carcinogen exposure (such as bracken fern toxicosis in cattle), actinic radiation, and rarely with papillomatosis.⁸In general, neoplastic diseases are rare in nonhuman primates, and SCC and lymphoma are the 2 most commonly reported oral neoplasms in these species.³ SCC have most commonly been reported in rhesus macaques (Macaca mullata) and baboons (Papio spp.) among nonhuman primate species.⁹ In rhesus macaques, SCC has occurred in the oral cavity,⁹ integument,^9,22 esophagus,⁹ stomach,²¹ lung,^9,13 prepuce–penis,¹⁰ cervix,⁹ uterus,⁹ and eye.⁹ These neoplasms have also been reported to occur in cynomolgus macaques,^14,15,17,19 marmosets, squirrel monkeys, tree shrews, capuchins, tamarins, black spider monkeys, sooty mangabies, a spectacled langur, and an orangutan.⁹ No report describing SCC in a pig-tailed macaque has been published previously. The oral cavity is the most common site of SCC in nonhuman primates, and metastasis occurs in approximately 23% of cases.⁹ The average age at diagnosis of oral SCC in rhesus macaques is 17.6 y.²² In baboons, SCC is the third most common neoplasm, after intestinal adenocarcinoma and lymphosarcoma.⁴ The following case report describes an oral SCC in a pregnant pig-tailed macaque.     

MiR-506 Suppresses Tumor Proliferation and Invasion by Targeting FOXQ1 in Nasopharyngeal Carcinoma

MiRNAs are small noncoding RNAs that play important roles in various biological processes including tumorigenesis. However, little is known about the expression and function of miR-506 in nasopharyngeal carcinoma (NPC). In this study, we showed that miR-506 was downregulated in nasopharyngeal carcinoma (NPC) cell lines and tissues. Ectopic expression of miR-506 dramatically suppressed cell proliferation, colony formation and invasion. Moreover, we identified the Forkhead box Q1 (FOXQ1) gene as a novel direct target of miR-506. MiR-506 exerts its tumor suppressor function through inhibition of the FOXQ1, which was involved in tumor metastasis and proliferation in various cancers. Furthermore, the expression of FOXQ1 is up-regulated in NPC cell lines and tissues. Taken together, our results indicate that miR-506 functions as a tumor suppressor miRNA in NPC and that its suppressive effects are mediated chiefly by repressing FOXQ1 expression.     

紫杉醇对人肝癌SMMC7721细胞NDRG1表达及增值的影响

旨在探索紫杉醇对人肝癌SMMC7721细胞NDRG1表达的影响,及紫杉醇对肝癌SMMC7721细胞系增殖的抑制作用。分别提取紫杉醇处理前后SMMC7721细胞的RNA,进行逆转录-聚合酶链反应(RT-PCR),判断紫杉醇处理前后肝癌细胞中NDRG1表达的情况;采用蛋白印迹技术(Western blotting)分析紫杉醇处理前后肝癌细胞中NDRG1蛋白表达的情况;应用不同浓度紫杉醇处理肝癌细胞,以MTT法检测处理前后肝癌细胞的抑制率,流式细胞术(FCM)观察细胞周期变化的况。结果表明紫杉醇处理后的肝癌SMMC7721细胞中NDRG1表达下降,紫杉醇浓度越高,NDRG1表达水平越低,具有浓度依赖性。以MTT法观察紫杉醇对肝癌细胞的抑制作用,试验结果表明不同浓度的紫杉醇处理肝癌SMMC7721细胞后,癌细胞被明显抑制;以流式细胞术观察紫杉醇作用后肝癌SMMC7721细胞周期的变化,结果显示G2-M期细胞比例升高的程度随浓度增高而升高,细胞越来越多地被阻滞在G2-M期,不能继续分裂增殖。分化相关基因NDRG1的表达可能是肝癌的发病机制之一,紫杉醇可抑制肝癌SMMC 7721细胞中NDRG1的表达;同时紫杉醇能使肝癌SMMC7721细胞的生长阻滞在G2-M期,从而显著抑制SMMC7721细胞的增殖,并且具有剂量、时间依赖效应。     

Multi-photon Imaging of Tumor Cell Invasion in an Orthotopic Mouse Model of Oral Squamous Cell Carcinoma

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient management strategies. Orthotopic mouse models of oral cancer have been developed to facilitate the study of factors that impact invasion and serve as model system for evaluating anti-tumor therapeutics. In these systems, visualization of disseminated tumor cells within oral cavity tissues has typically been conducted by either conventional histology or with in vivo bioluminescent methods. A primary drawback of these techniques is the inherent inability to accurately visualize and quantify early tumor cell invasion arising from the primary site in three dimensions. Here we describe a protocol that combines an established model for squamous cell carcinoma of the tongue (SCOT) with two-photon imaging to allow multi-vectorial visualization of lingual tumor spread. The OSC-19 head and neck tumor cell line was stably engineered to express the F-actin binding peptide LifeAct fused to the mCherry fluorescent protein (LifeAct-mCherry). Fox1^nu/nu mice injected with these cells reliably form tumors that allow the tongue to be visualized by ex-vivo application of two-photon microscopy. This technique allows for the orthotopic visualization of the tumor mass and locally invading cells in excised tongues without disruption of the regional tumor microenvironment. In addition, this system allows for the quantification of tumor cell invasion by calculating distances that invaded cells move from the primary tumor site. Overall this procedure provides an enhanced model system for analyzing factors that contribute to SCOT invasion and therapeutic treatments tailored to prevent local invasion and distant metastatic spread. This method also has the potential to be ultimately combined with other imaging modalities in an in vivo setting.     

人口腔鳞癌组织中HIF-1a和NF-kB的表达及意义

目的:探讨缺氧诱导因子(HIF-1a)和核因子-KB(NF-KB)口腔鳞癌中的表达及相互关系,研究它们的表达与肿瘤临床病理指标的联系.方法:应用SP染色法检测HIF-1a和NF-KB在49例口腔鳞癌组织、10例正常口腔黏膜组织中的阳性率.结果在口腔鳞癌中HIF-1a和NF-KB的阳性表达率分别为80.0%和78.4%,其阳性率及表达等级均显著高于正常对照组(P<0.05),在中一低分化组和有淋巴结转移组中的表达显著高于高分化组和无淋巴结转移组(P<0.05).HIF-1a表达与NF-KB表达成等级正相关(r=0.45,P<0.05).结论:HIF-1a或NF-KB与口腔鳞癌生物学行为有密切关系,二者的联合检测,有助于口腔鳞癌恶性程度和生物学特性的判断.     

High Levels of Nucleolar Spindle-Associated Protein and Reduced Levels of BRCA1 Expression Predict Poor Prognosis in Triple-Negative Breast Cancer

Purpose

Nucleolar spindle-associated protein (NuSAP1) is an important mitosis-related protein, and aberrant NuSAP1 expression is associated with abnormal spindles and mitosis. This study investigated the prognostic value of NuSAP1 in breast cancer.

Methods

Two sets of tissue microarrays (TMAs) that included samples from 450 breast cancer patients were constructed, of which 250 patients were training set and the other 200 patients were validation set. Immunohistochemical staining was performed to determine the NuSAP1 levels. A Kaplan-Meier analysis was used to estimate the prognostic value of NuSAP1 in breast cancer. A stepwise Cox analysis was performed to construct a risk-prediction model for triple-negative breast cancer (TNBC). All statistical analysis was performed with SPSS software.

Results

There were 108 (43.5%) and 88 (44.0%) patients expressed NuSAP1 in the training set and validation set respectively. High levels of NuSAP1 expression were related to poor disease-free survival (DFS) in both training (P = 0.028) and validation (P = 0.006) cohorts, particularly in TNBC. With combination of two cohorts, both NuSAP1 (HR = 4.136, 95% CI: 1.956–8.747, P < 0.001) and BRCA1 (HR = 0.383, 95% CI: 0.160–0.915, P = 0.031) were independent prognostic indicators of DFS in TNBC. A receiver operating characteristic (ROC) analysis revealed that the combination of NuSAP1 and BRCA1 significantly improved the prognostic power compared with the traditional model (0.778 versus 0.612, P < 0.001).

Conclusions

Our study confirms the prognostic value of NuSAP1 in breast cancer. The combination of NuSAP1 and BRCA1 could improve the DFS prediction accuracy in TNBC.