In vitro effect of iASPP on cell growth of oral tongue squamous cell carcinoma

iASPP is an inhibitory member of the apoptosis-stimulating proteins of P53 （ASPP） family. iASPP is over expressed in several malignant tumors and potentially affects cancer progression. However, the expression and potential role of iASPP in oral tongue squamous cell carcinoma （OTSCC） have not been addressed. In our study, we detected iASPP expression in OTSCC by immunohistochemistry, iASPP expression is up-regulated in OTSCC tissues. Moreover, in clinical pathology specimens, we found that increased iASPP expression correlates with poor differentiation and lymph node metastasis. Using multicellular tumor spheroids （MTS） and flow cytometry, we demonstrated that iASPP down-regulation arrests OTSCC cells at the G0/G1 phase, induces OTSCC cell apoptosis and inhibits OTSCC cell proliferation. These results indicate that iASPP plays a significant role in the progression of OTSCC and may serve as a biomarker or therapeutic target for OTSCC patients.     

E-cadherin、PDPN、TLR4在舌鳞状细胞癌中的表达及其与淋巴结转移的相关性研究

目的 研究E-cadherin、PDPN、TLR4在舌鳞状细胞癌中的表达及与淋巴结转移的相关性.方法 选取82例舌鳞状细胞癌患者作为研究对象,检测舌鳞状细胞癌组织以及癌旁正常组织内粘附分子E(E-cadherin)、平足蛋白(PDPN)、Toll样受体4(TLR4)的表达,同时与患者临床特征进行分析.结果 E-cadh...     

Sox2 nuclear expression is closely associated with poor prognosis in patients with histologically node-negative oral tongue squamous cell carcinoma

Sox2 is a marker of embryonic stem cell pluripotency and plays an important role in tumor progression. However, Sox2 expression in oral tongue squamous cell carcinoma (OTSCC) and its correlation with patients' prognosis have not been investigated so far. In this study, we detected the expression of Sox2 in 82 patients with histologically node-negative (pN0) OTSCC by immunohistochemistry, and evaluated its correlation with clinicopatologic factors and disease prognosis. Sox2 positive expression was detected in 62.2% patients and showed a significant association with large tumor size. Survival analysis showed that patients with Sox2 positive expression had significantly poorer overall, cancer-specific and disease-free survivals than those with Sox2 negative expression at 5years after operation (P=0.004, 0.004 and 0.001, respectively). Multivariate analysis demonstrated that Sox2 positive expression was an independent prognosticator of unfavorable overall, cancer-specific and disease-free survivals (P=0.032, 0.035 and 0.011, respectively). According to our results, Sox2 positive expression was frequent in pN0 OTSCC and involved in tumor progression. The measurement of Sox2 expression may be helpful in predicting relapse and prognosis of patients with pN0 OTSCC.     

TRB3 overexpression due to endoplasmic reticulum stress inhibits AKT kinase activation of tongue squamous cell carcinoma

Our investigation aims to evaluate the significance of TRB3, an endoplasmic reticulum stress (ERS)-inducible gene, and explore its relationship with AKT in oral tongue squamous cell carcinoma (OTSCC). Expression of TRB3 and phosphorylated AKT (p-AKT) in OTSCC tissues and adjacent normal tissues were assessed by RT-PCR, Western blot and immunohistochemistry assay. Correlation of TRB3 and AKT was validated by TRB3 adenovirus plasmid (Ad-TRB3) transfection and short hairpin RNA (shRNA) inhibition. The mRNA expression of TRB3 was significantly higher than adjacent noncancerous tissues by RT-PCR in 15 of 18 specimens of OTSCC (83.3%, P<0.01). Both of TRB3 and AKT were highly expressed in 13 of 18 (72.2%) specimens of OTSCC comparing with adjacent noncancerous tissues by Western blot assay (P<0.05). TRB3 was significantly elevated in 49.2% (63/128) of pathologically confirmed specimens and 13.3% (4/30) of adjacent noncancerous specimens by immunohistochemical analysis (P<0.01). TRB3 overexpression was closely correlated with tumor pathological T stage, lymph node metastasis and tumor recurrence. In addition, both mRNA and protein expression of TRB3 was increased under thapsigargin (TG) or tunicmycin (TU)-induced ERS in Tca8113 and CAL-27 cells. Moreover, expression of p-AKT protein decreased when Ad-TRB3 was transected with OTSCC Tca8113 cells. However, expression of p-AKT protein increased when TRB3 was inhibited by TRB3 shRNA inhibition. TRB3 expression was closely correlated with OTSCC prognosis. Under ERS, TRB3 was up-regulated, resulting in inhibiting the activation of AKT in OTSCC.     

PARP-1在舌鳞癌组织中的表达及意义

目的检测PARP-1在舌鳞癌中的表达水平,分析与舌鳞癌患者临床病理因素及预后的相关性,初步探讨PARP-1在舌鳞癌的预后价值。方法采用免疫组织化学法检测155例舌鳞癌组织及13例邻近正常舌组织的石蜡组织切片中PARP-1的表达水平,分析PARP-1的表达水平与舌鳞癌患者的临床病理因素及预后的相关性。结果155例舌鳞癌组织中,PARP-1阳性表达率为89.0%(138/155),其中58.1%(90/155)呈高表达,41.9%(65/155)呈低表达;13例邻近正常舌组织均无PARP-1表达,两者比较差异有统计学意义(χ²=64.815,P=0.000)。PARP-1的表达水平与患者的T分期(χ² =10.841,P=0.002)、N分期(χ²=8.962,P=0.007)、病理分期(χ²=17.151,P=0.000)呈正相关。PARP-1高表达者总生存时间(P=0.037)与无瘤生存时间(P=0.026)比低表达者短。结论PARP-1可能是促进舌鳞癌发生发展的癌基因,具有预测舌鳞癌预后的临床价值。     

Beclin －1在舌鳞癌组织中的表达及临床意义

目的：自噬基因 Beclin －1在肿瘤形成和进展过程中发挥重要作用,但其和舌鳞癌（OTSCC）的关系尚不清楚,本研究拟探讨 Beclin －1在舌鳞癌组织中的表达及其临床意义。方法：免疫组织化学法检测133例舌鳞癌组织中 Beclin －1蛋白水平,分析其表达水平与舌鳞癌患者临床病理特征及预后的相关性。结果：降低的 Beclin －1表达水平与 OTSCC 组织低分化、淋巴结转移、cTNM分期和更差的预后密切相关。结论：降低的Beclin －1表达水平与舌鳞癌组织低分化、淋巴结转移、cTNM 分期和更差的预后相关,提示 Beclin －1在OTSCC 发生和进展过程中发挥抑癌基因的功能。     

Ki-67、TOPⅡα在食管鳞状细胞癌组织中的表达及其与脉管侵犯的相关性

目的:探讨食管鳞状细胞癌组织中Ki-67、TOPⅡα的表达及其与脉管侵犯的相关性。方法:回顾性分析60例食管鳞状细胞癌患者的临床资料,采用免疫组织化学法检测肿瘤组织中Ki-67、TOPⅡα的表达,比较有脉管侵犯者和无脉管侵犯者上述分子标志物表达的差异。结果:46例(76.7％)患者有脉管侵犯,其中在侵犯深度、TNM分期、淋巴结转移及组织学分级方面差异具有统计学意义(P<0.05)。有脉管侵犯者的TOPⅡα、Ki-67的高表达率显著高于无脉管侵犯者(P<0.05)。亚组分析显示,有和无脉管侵犯者之间Ki-67高表达率的差异主要出现于男性、年龄≥65岁、肿瘤最大径＜5 cm、组织学G2-G3级、浸润深度T3及TNM分期Ⅰ-Ⅱ期;有和无脉管侵犯者之间TOPⅡα高表达率的差异主要出现于年龄≥65岁、肿瘤最大径≥5 cm及组织学G2-G3级。Spearman相关性分析显示,Ki-67 和TOPⅡα 两者在食管鳞状细胞癌组织中表达呈正相关（r=0.314,P＝0.015）。结论:有脉管侵犯食管鳞状细胞癌患者的Ki-67、TOPⅡα的阳性表达率显著高于无脉管侵犯者,提示二者在脉管侵犯中起着重要作用。     

MicroRNA-21 Promotes Oral Cancer Invasion via the Wnt/β-Catenin Pathway by Targeting DKK2

MicroRNA-21 (miR-21) is overexpressed in a wide variety of cancers and has been related to cellular proliferation, apoptosis, and invasion; however, the function of miR-21 is unknown in oral tongue squamous cell carcinoma (OTSCC). The purpose of this study was to examine miR-21 expression in OTSCC, correlate it with clinicopathological factors, and investigate its contribution to OTSCC cell invasion. MiR-21 expression in 79 primary OTSCCs was evaluated using locked nucleic acid in situ hybridization, and correlation was examined with the clinicopathological factors. To determine the miR-21 target, we searched for molecular genes involved in tumor invasion using the commonly cited prediction program miRanda. In an OTSCC cell line, SCC25 cells, we further evaluated whether miR-21 contributes to cell invasiveness by blocking its expression with a specific knockdown LNA probe and confirmed the direct target by Matrigel invasion assay and Western blotting. MiR-21 overexpression was detected in 60 of 79 cases (75.9 %) and correlated with the pattern of invasion (P?=?0.016). We selected DKK2 as a Wnt/antagonist involved in tumor invasion. MiR-21 overexpression was significantly correlated with the DKK2-/β-catenin- immunohistochemical phenotype. Knockdown of miR-21 significantly decreased the invasion potential of SCC25 cells with up-regulated DKK2. It was found that miR-21 is overexpressed and associated with tumor invasion in OTSCC, and that miR-21 promotes OTSCC cell invasion via the Wnt/β-catenin pathway by targeting DKK2 in vitro. These results suggest that miR-21 may be a potential therapeutic target for OTSCC treatment.     

PKM2蛋白在胃癌组织中的表达及其预后价值

目的 探讨M2型丙酮酸激酶（PKM2）在人胃癌组织中的表达,并分析其与胃癌临床病理特征的关系及预后价值。方法 构建含202例胃癌组织及其对应癌旁组织的组织芯片,采用免疫组化法检测胃癌组织中PKM2蛋白的表达,分析其阳性表达与临床病理参数和预后间的关系。结果PKM2在胃癌组织中的阳性表达率为62.4％（126／202）,高于癌旁组织的14.4％（29／202）,差异有统计学意义（P＜0.05）;胃癌组织中PKM2的阳性表达率与年龄、性别、肿瘤最大直径及分化程度均无关（P＞0.05）,与T分期、N分期及TNM分期有关,差异有统计学意义（P＜0.05）。PKM2表达阳性者的中位总生存期为42.9个月,短于阴性表达者的78.4个月（χ2=23.119, P＜0.001）,且PKM2阳性表达情况是影响胃癌预后的独立因素之一。结论PKM2蛋白在胃癌的发生和发展过程中起了重要作用,是评估胃癌患者预后的重要因素。     

ABCG2和p75NTR在食管鳞癌组织中的表达及其与临床病理学特征的关系

目的:观察ABCG2和p75NTR在食管鳞癌（esophageal squamous carcinoma cell,ESCC）组织及与癌旁正常组织中的表达差异及其与患者临床病理学特征及相互之间的关系。方法:以免疫组化方法检测80例食管鳞癌患者手术标本和62例癌旁正常食管组织中的ABCG2和p75NTR表达,分析其与临床病理学特征及生存率的关系。结果:ABCG2和 p75NTR在食管鳞癌组织中的表达显著高于癌旁正常食管组织;与TNM分期有显著相关性（P＜0.05）。结论:食管癌干细胞标志物ABCG2和p75NTR在食管鳞癌组织中高表达,并且高于癌旁组织,与TNM分期关系密切,提示该研究为探索食管癌干细胞标志物奠定了基础。     

lncRNA DNM3OS在喉鳞状细胞癌组织和细胞中的表达及其临床和生 物学意义

目的：检测lncRNA DNM3OS在喉鳞状细胞癌（laryngeal squamous cell carcinoma,LSCC）组织和LSCC细胞株中的表达及其临床意义,探讨其对LSCC TU177细胞体外增殖、迁移及侵袭的影响,并分析DNM3OS与EMT的关系。方法：从河北医科大学第四医院生物标本库选取2014年3月至2018年12月收治的68例LSCC患者手术切除的癌及癌旁组织标本,应用qPCR法检测DNM3OS在LSCC组织和细胞株中的表达水平。采用siRNA敲低TU177细胞中DNM3OS的表达,应用MTS、克隆形成及Transwell小室等方法分别检测敲低DNM3OS表达对TU177细胞增殖、迁移和侵袭等生物学行为的影响。应用qPCR和WB法检测转染si-DNM3OS后对EMT标志物上皮钙黏素（E-cadherin）、神经钙黏素（N-cadherin）、波形蛋白（vimentin）、扭曲蛋白（twist）、锌指转录因子2（SNAI2）mRNA和蛋白的变化。结果：LSCC组织中DNM3OS表达水平明显高于癌旁组织（P<0.01）,并与患者的TNM分期、淋巴结转移及生存期有关联（P<0.05 或 P<0.01）。DNM3OS在LSCC细胞株（Hep-2、AMC-HN-8、TU177、TU212及TU686）中均呈现不同程度的高表达（P<0.05 或 P<0.01）,转染si-DNM3OS后TU177细胞中DNM3OS的表达显著降低（P<0.01）。与对照组相比,DNM3OS表达敲低可抑制TU177细胞的体外增殖、迁移和侵袭能力（P<0.05 或 P<0.01）,可上调 TU177 细胞中E-cadherin的表达而下调 N-cadherin、vimentin、twist和SNAI2的表达（均P<0.01）。结论：DNM3OS高表达与LSCC的恶性进展有关,其可能为预测LSCC患者预后的潜在指标;DNM3OS可能通过影响EMT进程促进LSCC细胞的侵袭和转移。     

食管鳞癌组织中LncRNA SNHG7的表达水平及与患者预后的关系

目的:检测食管鳞癌肿瘤组织中长链非编码核仁小RNA宿主基因7（LncRNA SNHG7）表达水平,并探究其与食管鳞癌患者预后的关系。方法:选取2011年6月至2014年9月本院收治的食管鳞癌患者52例,采用实时定量PCR（qRT-PCR）法检测食管鳞癌肿瘤组织及癌旁组织中LncRNA SNHG7表达水平。结果:食管鳞癌肿瘤组织LncRNA SNHG7表达水平显著高于癌旁组织（P<0.05）;LncRNA SNHG7表达水平与患者临床分期、肿瘤分化程度、肿瘤体积、淋巴结转移有关（P<0.05）,与患者性别、年龄、肿瘤浸润深度无关（P>0.05）;LncRNA SNHG7高表达者术后生存率显著低于LncRNA SNHG7低表达者（P<0.05）;LncRNA SNHG7表达和肿瘤TNM分期是影响患者预后的独立危险因素。结论:LncRNA SNHG7在食管鳞癌肿瘤组织中高表达,是食管鳞癌术后不良预后的生物指标。     

钠尿肽受体A在食管鳞状细胞癌中的表达及对食管癌细胞迁移和侵袭能力的影响

目的 探讨钠尿肽受体A(NPRA)在食管鳞状细胞癌中的表达及其对食管癌细胞迁移和侵袭的影响.方法 采用免疫组织化学染色法检测119例食管鳞状细胞癌组织和91例癌旁组织中NPRA的表达情况,并分析食管鳞状细胞癌组织中NPRA表达情况与患者临床特征及预后的关系.采用蛋白质印迹法(Westernblot)检测食管癌细胞株Ec...     

The involvement of CHD5 hypermethylation in laryngeal squamous cell carcinoma

Chromodomain helicase DNA-binding protein 5 (CHD5) has been found to be a candidate tumor suppressor gene (TSG) in malignant neural tumors. In mice heterozygous for chd5 deficiency, the first tumor observed was pathological squamous cell carcinoma. More than 95% of primary laryngeal cancer is squamous cell carcinoma. Thus, we explored the expression of CHD5 in 65 patients with laryngeal squamous cell carcinoma (LSCC) using real-time PCR, immunohistochemistry and Western blotting. DNA methylation was detected using bisulfate-specific sequencing. The potential function of CHD5 was determined using MTT, apoptosis and transwell migration assays in CHD5-transfected Hep-2 cells. Our results revealed that the mRNA and protein expression levels of CHD5 in LSCC tissues were significantly lower than those in clear surgical margin tissues (p<0.05), and there is a significant correlation between the mRNA and protein expression levels of CHD5 (p<0.01). In addition, there were significant differences in CHD5 mRNA and protein levels with respect to the patient's clinical stage (p<0.05). Aberrant methylation of the CHD5 promoter was frequently found in the Hep-2 cell line and LSCC tumor tissues, especially tumor tissues from advanced TNM (p<0.05) or older patients (p<0.05). Finally, ectopic expression of CHD5 in laryngeal cancer cells led to significant inhibition of growth and invasiveness. Our data suggest that CHD5 is a tumor suppressor gene that is epigenetically downregulated in LSCC.     

钙通道蛋白α2δ1及miR-107在喉癌组织中的表达和临床意义

目的:检测钙通道蛋白α2δ1及microRNA-107(miR-107)在喉癌组织中的表达情况及相互作用关系,探讨它们在喉癌发病中可能发挥的作用。方法:收集40例喉癌患者的癌组织和临近正常组织,分别检测两种组织中α2δ1及其编码基因CACNA2D1和miR-107的表达情况,并且分析它们的表达情况是否与患者的临床特征及预后具有相关性,使用生物信息学方法寻找CACNA2D1和miR-107潜在结合位点,双荧光素酶报告基因去验证,并使用喉癌细胞TU212人为过表达和敲减miR-107后行细胞迁移和增殖实验。结果:免疫荧光和蛋白质印迹分析结果表明,喉癌组织中α2δ1的表达较高（P＜0.05）。RT-qPCR结果显示喉癌组织中miR-107的表达水平显著降低（P＜0.05）。miR-107的低表达与喉癌患者的淋巴结转移、肿瘤分化和原发部位有关（P＜0.05）,但与年龄和肿瘤TNM分期无关（P＞0.05）。CACNA2D1的高表达与喉癌患者的淋巴结转移、肿瘤分化和TNM分期密切相关（P＜0.05）,但与LSCC患者的年龄和肿瘤原发部位无关（P＞0.05）;CACNA2D1的表达水平与患者的复发和3年生存率有相关性（P＜0.05）,而miR-107与之无相关性（P＞0.05）;双荧光素酶报告基因实验证实miR-107与CACNA2D1的3'-UTR两个位点结合,在TU212细胞中过表达miR-107会抑制细胞的迁移和增殖,敲减后则增强喉癌细胞的迁移和增殖能力（P＜0.05）。结论:在喉癌患者体内,钙通道蛋白α2δ1扮演着促癌的作用,miR-107可以通过直接靶向调节CACNA2D1从而抑制喉癌细胞的迁移和增殖。     

p53 Expression Helps Identify High Risk Oral Tongue Premalignant Lesions and Correlates with Patterns of Invasive Tumour Front and Tumour Depth in Oral Tongue Squamous Cell Carcinoma Cases

Oral tongue squamous cell carcinoma (OTSCC) is the most common oral cancer subtype with a maximum propensity for regional spread. Our objective was to study if p53 expression might have any correlation with aggressive patterns of invasion within oral tongue cancers as well as with the histologically identified degree of oral tongue dysplasia. p53 immunoexpression was studied using immunohistochemistry in early staged OTSCCs (n=155), oral tongue dysplasias, (n=29) and oral tongue normal specimens (n=10) and evaluated for correlations with histological and clinicopathological parameters. Our study (n=194) showed a pattern of p53 expression increasing with different grades of tongue dysplasia to different grades of invasive OTSCC (p=0.000). Among the OTSCC tumours, positive p53 expression was seen in 43.2% (67/155) and a higher p53 labelling index was significantly associated with increased Bryne’s grade of the tumour invasive front (p=0.039) and increased tumour depth (p=0.018). Among the OTSCC patients with tobacco habits, (n=91), a higher p53 labelling index was significantly associated with increased risk of local recurrence (p=0.025) and with lymphovascular space involvement (p=0.014). Evaluation of p53 through varying degrees of dysplasia to oral tongue cancer indicates that p53 expression is linked to aggressive features of oral tongue cancers and tongue precancers entailing a closer monitoring in positive cases. Among the OTSCCs, p53 expression is associated with tumour aggressiveness correlating with increased grading of invasive tumour front and tumour depth.     

Decreased expression of claudin-3 is associated with a poor prognosis and EMT in completely resected squamous cell lung carcinoma

The deregulation of claudin-3 has been reported to correlate with the invasion and metastasis of various cancers, but little is known about its expression level and the prognostic value in squamous cell lung carcinoma (SqCC). The purpose of this study is to determine the expression levels and the prognostic value of claudin-3 in completely resected SqCC tissues, and the potential underlying mechanism. The protein expression of claudin-3, E-cadherin, β-catenin, and vimentin in the tumor tissues from 103 patients with surgically resected SqCC was examined using immunohistochemistry, western blots, as well as semi-quantitative estimation. The claudin-3 protein level was significantly associated with E-cadherin, β-catenin, and vimentin protein expression. A decreased claudin-3 protein level was significantly correlated with TNM stage, lymph node metastasis, and disease recurrence. Similarly, downregulation of E-cadherin was significantly correlated with lymph node metastasis and disease recurrence. Decreased β-catenin expression also had a significant correlation with disease recurrence. Univariate analyses indicated that the T stage, lymph node metastasis, the TNM stage, and the expression of claudin-3, β-catenin, and vimentin were significant predictors for overall survival (OS). Moreover, multivariate analyses demonstrated that the TNM stage and protein levels of claudin-3, β-catenin, and vimentin were independent predictors for OS of SqCC patients. Claudin-3 plays an important role in the epithelial–mesenchymal transition of SqCC and might be used as a potential prognostic factor for SqCC.