Effects of Damnacanthal and Nordamnacanthal on Proliferation,Apoptosis, and Migration of Oral Squamous Cell Carcinoma Cells

Cancer is one of the most common causes of death in the developed world, with one-third of people diagnosed with cancer during their lifetime. Oral cancer commonly occurs involving the buccal mucosa (cheeks), tongue, floor of the mouth and lip. It is one of the most devastating and disfiguring of malignancies. Morinda citrifolia L., commonly known as ‘noni’, belongs to the Rubiaceae family. It is native to the Pacific islands, Hawaii, Caribbean, Asia and Australia. The plant displays broad curative effects in pharmacological studies. Damnacanthal (DAM) and Nordamnacanthal (NDAM), anthraquinone compounds isolated from the roots of Morinda citrifolia L., has been used for the treatment of several chronic diseases including cancer. The objectives of this study were to evaluate cytotoxicity, morphological changes, cell death mode (apoptosis/necrosis), and cell migration induced by DAM and NDAM on the most common type of oral cancer, oral squamous cell carcinoma (OSCC)cells. Anti-proliferative effects of these compounds against OSCC cell lines were determined by MTT assay. The mode of cell death was analysed by phase contrast and fluorescent microscopy as well as flow cytometry. In addition, cell migration was assessed. The results showed that DAM and NDAM exerted cytotoxicity against OSCC cells with IC50 values of 1.9 to >30 μg/ml after 72 h treatment. Maximum growth inhibition among the tested cell lines for both compounds was observed in H400 cells, and thus it was selected for further study. The study demonstrated inhibition of H400 OSCC cell proliferation, marked apoptotic morphological changes, induction of early apoptosis, and inhibition of cell migration by DAM and NDAM. Therefore, this information suggests that these compounds from noni have potential for used as anti tumor agents for oral cancer therapy.     

食管鳞癌中LncRNA DLEU1的表达及对食管鳞癌细胞增殖和迁移的影响

目的 研究长链非编码RNA淋巴细胞白血病缺失基因1(DLEU1)在食管鳞状细胞癌(ESCC)组织中的表达,及其对肿瘤细胞增殖和迁移的影响.方法 收集58例手术切除的ESCC及其相应癌旁组织,RT-qPCR检测ESCC组织和细胞中DLEU1的表达水平.Log rank Test法分析ESCC组织中DLEU1的表达与患者临...     

miR-100对食管鳞癌细胞株Ec-109的抑制作用

目的 探讨miR-100对食管鳞癌细胞的调控作用。方法 构建携带GFP的miR-100表达质粒,脂质体转染食管鳞癌细胞株Ec-109。分别利用流式细胞仪、细胞划痕和Transwell实验检测miR-100对细胞周期、凋亡和迁移的调节作用。结果 在食管鳞癌细胞中高表达miR-100可诱导G₁期阻滞,即停留于G₁期的细胞数量增加,进入S和G₂/M期的数量减少;在无血清培养的条件下miR-100的高表达可促进细胞凋亡同时抑制食管鳞癌细胞迁移。结论 miR-100在食管鳞癌细胞株Ec-109中高表达可诱导细胞周期G₁期阻滞、抑制细胞的迁移并促进其凋亡。     

食管鳞癌中p53、PCNA和EGFR的表达与化疗疗效的关系

目的:探讨p53、PCNA和EGFR在食管鳞癌中的表达情况与化疗疗效的关系。方法:对66例食管鳞癌化疗前的活检标本用免疫组化技术分别检测p53、PCNA、EGFR的表达。结果:p53蛋白积聚阳性组化疗有效率(16.7%)明显低于p53蛋白积聚阴性组(70.8%)(P<0.01),PCNA过表达组化疗有效率(79.4%)高于PCNA弱表达组(31.3%)(P<0.01),EGFR过表达组化疗有效率(30.4%)低于弱表达组(69.8%)(P<0.01)。经Logistic回归分析,3个指标中,PCNA对化疗疗效的预测价值较大(P<0.01)。结论:食管鳞癌中p53、PCNA和EGFR的表达情况对化疗疗效均有一定的预测价值。其中PCNA价值较大。     

p16蛋白在食管鳞癌中的表达及其意义

目的：研究p16蛋白表达与食管鳞癌生物学行为的关系。方法：应用免疫组化LSAB法检测52例食管鳞癌p16蛋白表达。结果：21例食管鳞癌p16蛋白表达阳性。阳性率为40.4%，随着恶性程度增加及病程进展，p16蛋白阳性率逐渐下降；p16蛋白阳性与淋巴结转移有关。p16蛋白阴性患者死亡率明显高于p16蛋白阳性者，且存活时间短。结论：检测食管鳞癌组织p16蛋白表达，有助于综合判断食管鳞癌恶性程度，转移潜能和预后。     

下调HDAC6表达对食管鳞癌细胞周期、细胞迁移的影响及其分子机制探讨

目的:探讨组蛋白去乙酰化酶6（HDAC6）表达的下调对食管鳞癌细胞周期、细胞迁移的影响及其分子机制。方法:将食管鳞癌EC9706细胞分为三组,即未处理组、对照siRNA组和HDAC6 siRNA组,后两组分别转染对照siRNA和HDAC6 siRNA。采用半定量反转录-聚合酶链反应、蛋白印迹方法检测各组细胞中细胞增殖和周期相关因子p21 mRNA和蛋白以及细胞迁移相关因子E-cadherin mRNA和蛋白的表达水平的变化。结果:HDAC6 siRNA组中p21及E-cadherin mRNA和蛋白的表达水平分别是0.440±0.120、0.840±0.070和0.580±0.090、0.450±0.080,且均明显高于未处理组（0.165±0.090、0.090±0.020;0.088±0.009、0.054±0.011）和对照siRNA组（0.163±0.021、0.070±0.040;0.820±0.070、0.066±0.007）中p21及E-cadherin mRNA和蛋白的表达水平,其表达差异均具有统计学意义（P均<0.05）,而未处理组和对照siRNA组之间p21及E-cadherin mRNA和蛋白的表达无差异（P＞0.05）。结论:HDAC6表达下调对食管鳞癌细胞周期的影响可能与p21表达的升高相关;对细胞迁移的影响可能与E-cadherin表达的上调有关。     

HDAC6 siRNA Inhibits Proliferation and Induces Apoptosis of HeLa Cells and its Related Molecular Mechanism

Objective: To investigate the effects of histone deacetylase 6 (HDAC6) siRNA on cell proliferation and cellapoptosis of the HeLa cervical carcinoma cell line and the molecular mechanisms involved. Methods: Divisionwas into three groups: A, the untreated group; B, the control siRNA group; and C, the HDAC6 siRNA group.Lipofectamine 2000 was used for siRNA transfection, and Western blot analysis was used to determine the proteinlevels. Cell proliferation and apoptosis were characterized using a CCK-8 assay and flow cytometry, respectively.Results: HDAC6 protein expression in the HDAC6 siRNA-transfection group was significantly lower (P < 0.05)than in the untreated and control siRNA groups. The CCK-8 kit results demonstrated that the proliferation ofHeLa cells was clearly inhibited in the HDAC6 siRNA transfection group (P < 0.05). In addition, flow cytometryrevealed that the early apoptotic rate (26.0% ± 0.87%) was significantly elevated (P < 0.05) as compared withthe untreated group (10.6% ± 1.19%) and control siRNA group (8.61% ± 0.98%). Furthermore, Western blotanalysis indicated that bcl-2 protein expression in the HDAC6 siRNA–transfection group was down-regulated,whereas the expression of p21 and bax was up-regulated. Conclusion: HDAC6 plays an essential role in theoccurrence and development of cervical carcinoma, and the down-regulation of HDAC6 expression may beuseful molecular therapeutic method.     

Network Analyses of Gene Expression following Fascin Knockdown in Esophageal Squamous Cell Carcinoma Cells

Fascin-1 (FSCN1) is an actin-bundling protein that induces cell membrane protrusions, increases cell motility,and is overexpressed in various human epithelial cancers, including esophageal squamous cell carcinoma (ESCC).We analyzed various protein-protein interactions (PPI) of differentially-expressed genes (DEGs), in fascinknockdown ESCC cells, to explore the role of fascin overexpression. The node-degree distributions indicated thesePPI sub-networks to be characterized as scale-free. Subcellular localization analysis revealed DEGs to interactwith other proteins directly or indirectly, distributed in multiple layers of extracellular membrane-cytoskeleton/cytoplasm-nucleus. The functional annotation map revealed hundreds of significant gene ontology (GO) terms,especially those associated with cytoskeleton organization of FSCN1. The Random Walk with Restart algorithmwas applied to identify the prioritizations of these DEGs when considering their relationship with FSCN1. Theseanalyses based on PPI network have greatly expanded our comprehension of the mRNA expression profilefollowing fascin knockdown to future examine the roles and mechanisms of fascin action.     

食管鳞癌中p16基因启动子区甲基化及其表达

 目的探讨食管鳞癌（ESCC）p16基因甲基化的状况及其表达与食管鳞癌临床病理特征之间的关系。方法采用甲基化特异性PCR方法（MSP）分别检测75例食管癌组织、癌旁组织和切缘组织p16基因启动子区域CpG岛甲基化状态。采用Envision免疫组化法检测食管癌组织及癌旁组织的p16蛋白的表达。结果75例标本中，食管癌组织、癌旁组织和切缘细织p16基因甲基化率分别为41．3％（31／75）、13．3％（10／75）和6．67％（5／75）。癌组织和癌旁组织P16蛋白的阳性表达率分别为29．3％（22／75）和56．7％0（17／30）。31例癌组织p16基因甲基化阳性标本中有2例（6．4％）检测到P16蛋白的表达，而44例癌组织p16基因甲基化阴性标本中有20例（45．5％）检测到P16蛋白的表达。食管癌组织p16基因甲基化率显著高于癌旁组织和切缘组织（P〈0.01），P16蛋白表达与p16基因甲基化呈负相关。p16基因启动子区甲基化与食管癌的组织学分级、肿瘤部位无明显相关，与临床分期、淋巴转移密切相关。结论p16基因甲基化在食管癌发生发展中起着重要作用，食管鳞癌的分期和淋巴结转移与p16基因甲基化之间有密切关系。     

缺锌饮食对小鼠食管鳞状上皮细胞增殖的影响及机制

目的 探讨饮食锌缺乏对小鼠食管鳞状上皮细胞增殖的影响及其可能机制.方法 采用锌-小鼠-食管模型,将C57BL/6小鼠随机分为两组,每组10只,分别以足锌饲料和缺锌饲料喂养,观察小鼠体质量变化、血液和组织中的锌含量变化以及小鼠食管黏膜组织中增殖细胞核抗原(PCNA)、P38丝裂原活化蛋白激酶(P38MAPK)、核因子κB...     

HOXB7基因敲除对人肝癌细胞的增殖、侵袭及迁移能力的影响

目的 研究同源异型盒基因HOXB7（Homebox7）对人肝癌细胞增殖、侵袭、迁移能力的影响及部分机制。方法 采用荧光定量PCR法选取HOXB7 mRNA高表达的人肝癌SMMC-7721细胞和HepG2细胞;设计靶向HOXB7的shRNA,瞬时转染肝癌SMMC-7721细胞,荧光定量PCR和Westernblot法检测shRNA靶向沉默的效果,CCK8、Transwell和划痕实验分别用于检测细胞增殖、侵袭和迁移能力;Western blot法测量肝癌SMMC-7721细胞中Smad3、p-Smad3表达水平。结果 HOXB7 shRNA转染的肝癌SMMC-7721细胞中HOXB7基因mRNA和蛋白表达水平均显著降低,同时细胞增殖、侵袭和迁移能力随之下降,且TGF-β通路中p-Smad3蛋白表达水平下调。结论 HOXB7可促进肝癌细胞增殖、侵袭以及迁移能力,其机制可能与调节Smad3磷酸化水平有关。     

CYFRA21—1检测在食管鳞癌中的临床应用价值

目的 研究细胞角蛋白19可溶性片段（CYFRA21－1）在食管鳞癌中的浓度变化及临床意义。方法 采用免疫放射法（IRMA）检测35例食管鳞癌患者血浆中的CYFRA21－1、SCC抗原及CEA的浓度,研究其浓度与其临床病理因素的相关性;比较27例手术治疗患者术前、术后第1天和术后第7天的浓度变化。结果 在35例食管鳞癌患者中,20例CYFRA21－1阳性（＞3.5ng/ml）,其特异性、敏感性及正确性分别为100.0%,57.1%,66.7%,联合检测CYFRA21－1及SCC抗原,其敏感性可达77.1%;术前与术后相比,CYFRA21－1血浆浓度有非常显著性差异（P＜0.01）,术后CYFRA21－1血浆浓度呈梯度下降;CYFRA21－1血浆浓度与食管鳞癌的临床病理因素相关（肿瘤大小、肿瘤侵犯程度、淋巴结转移及切除率）。结论 CYFRA21－1是食管鳞癌的有效肿瘤标志物,联合检测CYFRA21－1和SCC抗原对于判断肿瘤的复发和转移具有重要的临床意义,CYFRA21－1可作为判断食管鳞癌患者预后的重要指标之一。     

Antitumor Effects of Fucoidan Via Apoptotic and Autophagic Induction on HSC-3 Oral Squamous CellCarcinoma

Objective: Many studies suggested that fucoidan has anticancer potential. The objective of the present study was to determine the cytotoxic effects and mechanism of cell death induced by fucoidan extracted from Fucus vesiculosus on HSC-3 oral squamous cell carcinoma. Methods: HSC-3 cells were treated with 0, 100, 200, and 400 μg/mL of fucoidan. Cell viability was measured using MTT assay. Apoptosis and cell cycle were measured with a flow cytometry-based assay. Chromatin condensation and nuclear fragmentation were determined using Hoechst 33342 staining. Mitochondrial membrane potential (ΔΨm) was determined using the JC-1 kit. The apoptotic, anti-apoptotic, and autophagic markers study were done by western blot analysis. Results: the viable cell number of treated HSC-3 cells was decreased. Moreover, treated cells were arrested in the G0/G1 phase. Annexin V/PI staining revealed that fucoidan could induce apoptosis in HSC-3 cells. Western blot analysis suggested the up-regulation of apoptotic markers including cleaved caspase-3, cleaved PARP, Bax, and autophagic markers including LC3-II and Beclin-1 but down-regulation of anti-apoptotic markers, Bcl-2. Fucoidan could disturb ΔΨm and induce chromatin condensation with nuclear fragmentation. Conclusion: fucoidan has potential in anticancer properties against HSC-3 cells manifested by the induction of apoptosis, cell cycle arrest, and autophagy.     

人端粒酶hTRT在癌旁黏膜上皮和食管鳞癌组织中的表达及其意义

目的 研究食管黏膜鳞状上皮和食管鳞癌组织中端粒酶hTRT的表达，并探讨其与食管癌发生、发展的关系。方法 应用免疫组化S—P法，观察10例正常食管黏膜、35例癌旁食管黏膜上皮，120例食管癌组织微阵列中端粒酶hTRT的表达情况。结果 正常食管黏膜鳞状上皮细胞末见端粒酶hTRT阳性表达，癌旁黏膜上皮和癌组织端粒酶hTRT阳性表达串分别为94．3％和81．7％。癌组织hTRT的表达与癌组织的组织学分级、浸润深度和淋巴结转移无关(P＞0．05)。结论 端粒酶hTRT的激活表达与食管黏膜上皮的恶性转化密切相关，端粒酶hTRT的重新激活可能在食管癌的组织发生中起关键性作用。     

食管鳞状细胞癌组织中Ezrin基因的表达和临床意义

目的探讨食管鳞状细胞癌组织及正常食管黏膜组织中埃兹蛋白（Ezrin）的表达情况,并分析其与食管鳞状细胞癌临床病理及患者5年生存率的关系。方法203例食管癌患者均来自河南林州市。利用组织微阵列技术制作组织芯片,采用免疫组织化学ABC法分析203例食管鳞癌组织（观察组）和相应正常食管黏膜组织（对照组）中Ezrin蛋白的表达情况。结果观察组中Ezrin蛋白表达率（67.0%）明显高于对照组（38.5%）（P<0.05）;Ezrin蛋白高表达与淋巴结转移、TNM分期有关,而与年龄、性别、肿瘤细胞分化程度和大体分型无关;Ezrin蛋白高表达的食管鳞癌患者的5年生存率明显低于未高表达者。结论Ezrin蛋白的异常表达可能与食管鳞状细胞癌的发生、发展相关,检测Ezrin可能为食管癌预后判断提供依据。     

食管鳞状细胞癌组织中miRNA-375的表达及其临床意义

目的 探讨miRNA-375表达与食管鳞状细胞癌临床病理特征的关系.方法 通过实时荧光定量PCR,检测miRNA-375在67例食管癌及癌旁正常组织的表达.结果 采用循环阈值(△Ct)对miRNA-375在食管癌和癌旁正常组织中表达量进行定量分析,食管癌组织△Ct为9.61±1.52,对应正常组织△Ct为10.86±1.44,其中有80.6％的食管癌组织其表达量低于癌旁正常组织(2-△△Ct＜1).结论 miRNA-375低表达可能与食管鳞状细胞癌的发生、发展、转移相关.     

Transmembrane Protein 166 Expression in Esophageal Squamous Cell Carcinoma in Xinjiang,China

Objective: Transmembrane protein 166 (TMEM166) expression in esophageal squamous cell carcinoma(ESCC) and remote normal esophageal tissues was examined to assess any role in tumour biology. Methods:TMEM166 mRNA expression in 36 cases with ESCC (36 tumour samples, 36 remote normal esophageal tissuesamples) was detected by RT-PCR. TMEM166 protein expression was analysed in paraffin-embedded tissuesamples from the same cases by immunohistochemistry. Results: Semi-quantitative analysis showed TMEM166mRNA expression in ESCCs to be significantly lower than in remote normal esophageal tissues (0.759±0.713 vs.2.622±1.690, P=0.014). TMEM166 protein expression was also significantly reduced (69.4% vs. 94.4%, P<0.01).Conclusion: TMEM166 mRNA and protein expression demonstrated significant reduction in ESCCs comparedwith remote esophageal tissues, albeit with no correlation with tumour size, differentiation, stage, and lymphnode metastasis, suggesting a role in regulating autophagic and apoptotic processes in the ESCC.     

Expression and Clinical Significance of REPS2 in Human Esophageal Squamous Cell Carcinoma

Objective: REPS2 plays important roles in inhibiting cell proliferation, migration and in inducing apoptosisof cancer cells, now being identified as a useful biomarker for favorable prognosis in prostate and breast cancers.The purpose of this study was to assess REPS2 expression and to explore its role in esophageal squamous cellcarcinoma (ESCC). Methods: Protein expression of REPS2 in ESCCs and adjacent non-cancerous tissues from 120patients was analyzed by immunohistochemistry and correlated with clinicopathological parameters and patientoutcome. Additionally, thirty paired ESCC tissues and four ESCC cell lines and one normal human esophagealepithelial cell line were evaluated for REPS2 mRNA and protein expression levels by quantitative RT-PCR andWestern blotting. Results: REPS2 mRNA and protein expression levels were down-regulated in ESCC tissuesand cell lines. Low protein levels were significantly associated with primary tumour, TNM stage, lymph nodemetastasis and recurrence (all, P < 0.05). Survival analysis demonstrated that decreased REPS2 expression wassignificantly associated with shorter overall survival and disease-free survival (both, P < 0.001), especially inearly stage ESCC patients. When REPS2 expression and lymph node metastasis status were combined, patientswith low REPS2 expression/lymph node (+) had both poorer overall and disease-free survival than others (both,P < 0.001). Cox multivariate regression analysis further revealed REPS2 to be an independent prognostic factorfor ESCC patients. Conclusions: Our findings demonstrate that downregulation of REPS2 may contribute tomalignant progression of ESCC and represent a novel prognostic marker and a potential therapeutic target forESCC patients.