靶向转录因子NF-κB p65的siRNA对Hep-2细胞增殖及表达的影响

目的:以喉鳞状细胞癌Hep-2细胞为研究对象,应用RNAi技术特异性的抑制NF-κBp65的表达,观察其对癌细胞增殖、蛋白和mRNA表达的影响.方法:用NF-κBp65siRNA转染喉鳞状细胞癌Hep-2细胞,采用Western Blotting方法检测Hep-2细胞转染前、后NF-κBp65的蛋白表达;采用RT-PCR检测Hep-2细胞转染前、后NF-κBp65的mRNA表达水平;采用噻唑蓝(MTT)比色法检测细胞增殖情况.结果:Western Blotting结果显示随着NF-κBp65siRNA作用时间的延长,蛋白表达水平逐渐降低.半定量RT-PCR结果表明,转染NF-κBp65siRNA 24、48和72 h后的Hep-2细胞,与未转染组的Hep-2细胞相比,NF-κBp65 mRNA的表达水平逐渐下调(P＜0.05);MTT法显示不同时间NF-κBp65siRNA转染Hep-2细胞后,细胞增殖活性受抑制情况与对照组比较差异有统计学意义(P＜0.05).结论:NF-κBp65siRNA对Hep-2细胞的增殖活性有明显的抑制作用,并且可以特异性地抑制目的基因NF-κB p65在mRNA和蛋白水平上的表达,其作用具有时间依赖性.     

全身应用激素对鼻息肉组织中NF-κBp65及I κ Bα表达的影响

目的观察全身应用糖皮质激素对鼻息肉组织中NF-κBp65、IκBα表达的影响。方法采用免疫组化SP法,检测在激素治疗前后NF-κBp65、IκBα在鼻息肉组织中的表达,运用HPIAS-2000高分辨率图像分析系统分别对激素治疗前后鼻息肉组织的黏膜上皮细胞、腺上皮细胞、血管内皮细胞、炎性细胞进行光密度测定。结果糖皮质激素治疗前,NF-κBp65阳性表达主要分布在鼻息肉组织的黏膜上皮细胞、炎性细胞、腺上皮细胞及血管内皮细胞的胞浆和部分胞核中;IκBα阳性表达主要分布在鼻息肉组织的黏膜上皮细胞、炎性细胞、腺上皮细胞及血管内皮细胞的胞浆中。激素治疗后,IκBα在鼻息肉组织黏膜上皮细胞炎性细胞中表达显著增高,差异具有显著性(P<0.05);NF-κBp65在鼻息肉组织黏膜上皮细胞,炎性细胞中表达显著降低,差异具有显著性(P<0.05)。结论糖皮质激素促进鼻息肉组织黏膜上皮细胞、炎性细胞中IκBα的表达,抑制黏膜上皮细胞、炎性细胞中NF-κBp65的表达,可能是其治疗鼻息肉的作用机制之一。     

核转录因子κ-Bp65在中耳胆脂瘤上皮中的表达和意义

目的检测核转录因子-κBp65（nuclear factor kappa Bp65,NF-κBp65）在中耳胆脂瘤上皮中的表达和活化,探讨其在中耳胆脂瘤发病机制中的可能作用。方法采用免疫组织化学SP法检测30例中耳胆脂瘤组织标本与15例正常外耳道皮肤标本中NF-κBp65蛋白的表达。结果NF-κBp65蛋白阳性表达定位于上皮细胞核。NF-κBp65蛋白在中耳胆脂瘤上皮组织中阳性表达率为63.3%,明显高于正常外耳道皮肤组的20.0%（P〈0.01）。结论NF-κBp65蛋白在中耳胆脂瘤上皮的异常表达可能在胆脂瘤的发生、发展过程中起重要作用。胆脂瘤上皮中NF-κBp65的活化可能参与了胆脂瘤上皮细胞过度增殖机制。     

热休克蛋白27与NF-κBp65在喉鳞状细胞癌中的基因表达及临床意义

目的研究热休克蛋白27（heat shock protein27,HSP27）及核因子NF-κBp65（NF-κBp65）在喉癌及癌旁组织中基因表达水平,并探讨两者与喉癌临床病理因素的相关性。方法采用RT-PCR方法分别检测HSP27 mRNA和NF-κBp65 mRNA在喉癌组织及癌旁组织中的表达水平,并统计两者与喉癌不同临床病理因素的关系。结果 HSP27 mRNA和NF-κBp65 mRNA分别在喉癌组织中的表达高于癌旁组织,两基因与喉癌不同病理分型有相关性,与临床分期及有无淋巴结转移无关,且NF-κBp65 mRNA和HSP27 mRNA在喉癌中呈负相关性（r=-0.4367,P〈0.05）。结论 HSP27 mRNA和NF-κBp65mRNA表达水平随喉癌恶性程度的增高HSP27 mRNA表达量逐渐下降,而NF-κBp65 mRNA表达量逐渐升高,推测HSP27在人喉癌中可能负性调节NF-κB信号传导通路,HSP27有望成为喉鳞癌基因治疗新的作用靶点。     

全身应用激素对鼻息肉组织中NF-κBp65及I κ Bα表达的影响

目的观察全身应用糖皮质激素对鼻息肉组织中NF-κ Bp65、I κ B α表达的影响.方法采用免疫组化SP法,检测在激素治疗前后NF-κ Bp65、Iκ B α在鼻息肉组织中的表达,运用HPIAS-2000高分辨率图像分析系统分别对激素治疗前后鼻息肉组织的黏膜上皮细胞、腺上皮细胞、血管内皮细胞、炎性细胞进行光密度测定.结果糖皮质激素治疗前,NF-κBp65阳性表达主要分布在鼻息肉组织的黏膜上皮细胞、炎性细胞、腺上皮细胞及血管内皮细胞的胞浆和部分胞核中;I κ B α阳性表达主要分布在鼻息肉组织的黏膜上皮细胞、炎性细胞、腺上皮细胞及血管内皮细胞的胞浆中.激素治疗后,I κ B α在鼻息肉组织黏膜上皮细胞,炎性细胞中表达显著增高,差异具有显著性(P＜0.05);NF-κ Bp65在鼻息肉组织黏膜上皮细胞,炎性细胞中表达显著降低,差异具有显著性(P＜0.05).结论糖皮质激素促进鼻息肉组织黏膜上皮细胞、炎性细胞中IκB α的表达,抑制黏膜上皮细胞、炎性细胞中NF-κBp65的表达,可能是其治疗鼻息肉的作用机制之一.     

EGCG对鼻咽癌细胞增殖抑制和凋亡诱导时NF-κB和caspase-3的表达变化

目的探讨表没食子儿茶素没食子酸酯（epigallocatechin-3-gallate,EGCG）作用于鼻咽癌CNE-2细胞后,对转录因子NF-κB和凋亡效应酶caspase-3表达的影响,以及这两个基因的表达与鼻咽癌细胞凋亡和增殖的关联性。方法体外培养鼻咽癌细胞CNE-2,MTT法检测细胞增殖的抑制作用;Hoechst染色检测细胞凋亡;流式细胞仪分析细胞周期的改变。RT-PCR检测NF-κB和caspase-3的表达。结果EGCG能明显的抑制鼻咽癌细胞增殖,诱导其凋亡,并表现为浓度依赖性：流式分析显示,EGCG干预鼻咽癌细胞CNE-2在48小时后,细胞被阻滞于G1期。80mg/L EGCG处理鼻咽癌细胞24小时和48小时后,均能下调NF-κB表达,上调caspase-3表达。结论EGCG可能通过抑制NF-κB和诱导caspase-3的表达来发挥其抗鼻咽癌细胞的生物学作用。     

克拉霉素对离体鼻息肉组织中环氧合酶-2及核因子κB表达的影响

目的：检测用不同浓度梯度的克拉霉素干预后,离体鼻息肉组织中环氧合酶-2（COX-2）及核因子κB（NF-κB）的表达状况,探讨其在鼻黏膜炎症机制中的作用。方法：将鼻息肉组织块分别与克拉霉素（0、10^-6、10^-5及10^-4mol／L）于体外共同培养1d,应用Western blot和荧光实时定量PCR技术检测COX-2和NF-κB亚基的表达水平,观察克拉霉素干预后COX-2和NF-κB表达的变化规律。结果：对照组（0mol／L克拉霉素组）鼻息肉组织中COX-2、NF-κBp50和NF-κcBp65的蛋白质和核酸表达水平最高;随着克拉霉素干预剂量的增加,COX-2、NF-κBp50和NF—κBp65的表达水平呈剂量依赖性下降。相关分析表明,同一组鼻息肉组织中,COX-2核酸表达量分别与NF-κBp50、NF-κBp65呈直线正相关。结论：COX-2异常表达参与了鼻-鼻窦黏膜慢性炎症反应,克拉霉素可能通过阻断NF-κB通路来下调COX-2的合成,发挥其抗炎作用。     

NF-κB抑制剂对慢性鼻咽炎豚鼠鼻咽黏膜炎症细胞增殖与凋亡活性的影响

目的 观察核转录因子(nuclear factor-kappa B,NF-κB)特异性抑制剂吡咯二硫代氨基甲酸(pyrrolidine dithiocarbamate,PDTC)对慢性鼻咽炎豚鼠鼻咽黏膜组织浸润炎症细胞增殖与凋亡活性的影响,探讨慢性鼻咽炎在鼻咽癌前病变发生与发展过程中的地位.方法 利用细菌病毒联合法建立豚鼠慢性鼻咽炎模型,然后将模型鼠随机分为慢性鼻咽炎对照组和PDTC干预组,并平行设置正常对照组.经药物处理后,取各组动物鼻咽黏膜组织观察病理组织学改变,免疫组化法检测鼻咽黏膜NF-κB p65和增殖细胞核抗原(PCNA)表达活性,末端转移酶介导的dUTP缺口末端标记(TUNEL)技术检测浸润炎症细胞凋亡指数.结果 模型组鼻咽黏膜组织浸润炎性细胞NF-κB p65表达活性明显高干正常对照组(P<0.01),PDTC干预组则明显低于模型组;模型组浸润炎症细胞凋亡时间延长、增殖活性升高,PDTC干预组炎症细胞凋亡活性、增殖水平与正常对照组的差异无统计学意义.结论 NF-κB抑制剂PDTC对慢性鼻咽炎黏膜组织中浸润炎症细胞的增殖活性显示明显抑制效应,其机制可能是通过促进靶细胞凋亡活性而发挥效应,提示鼻咽炎有演变为鼻咽癌前病变的潜在风险.     

核转录因子κB p65在喉癌中的表达及临床意义

目的：探讨核转录因子κBp65（NF-κBp65）在喉癌中的表达及临床意义。方法：采用免疫组织化学及原位杂交方法,检测48例喉癌组织和20例正常喉黏膜组织中NF-κBp65的表达水平及与肿瘤生物学行为间的关系。结果：①NF-κBp65的表达在喉癌与正常喉黏膜组织间的差异有统计学意义（P〈0.05）。②NF-κBp65的高表达与颈淋巴结转移和临床分期有相关性,而与肿瘤部位及病理分化无相关性。结论：NF-κBp65是判断喉癌预后极具价值的指标。     

NF-κBp65、IκBα在人鼻息肉组织中的表达

目的通过检测鼻息肉组织、慢性鼻-鼻窦炎病人钩突黏膜及鼻中隔偏曲病人下鼻甲黏膜中NF-κBp65、IκBα的表达,探讨NFκ-Bp65、IκBα在鼻息肉、慢性鼻-鼻窦炎发病中所起的作用。方法采用免疫组化SP法检测NFκ-Bp65、IκBα在30例鼻息肉组织、20例慢性鼻-鼻窦炎病人单侧钩突黏膜及20例鼻中隔偏曲病人下鼻甲黏膜中的表达及分布,运用HPIAS-2000高分辨率图像分析系统分别对3种组织的黏膜上皮细胞、腺上皮细胞、血管内皮细胞、炎性细胞进行光密度测定。结果①NF-κBp65阳性表达主要分布在鼻息肉组织的黏膜上皮细胞、炎性细胞、腺上皮细胞及血管内皮细胞的胞浆和部分胞核中。②IκBα阳性表达主要分布在鼻息肉组织的黏膜上皮细胞、炎性细胞、腺上皮细胞及血管内皮细胞的胞浆中。③NFκ-Bp65在鼻息肉组、钩突组及下鼻甲组黏膜上皮细胞、炎性细胞、腺上皮细胞及血管内皮细胞中表达依次降低,且差异具有统计学意义。④IκBα在鼻息肉组、钩突组及下鼻甲组组织黏膜上皮细胞、炎性细胞、腺上皮细胞及血管内皮细胞的表达依次降低,且差异具有统计学意义。结论NF-κBp65、IκBα可能参与鼻息肉和慢性鼻-鼻窦炎的炎症反应过程。     

Ganglion cell loss continues during hair cell regeneration

Hair cells and ganglion cells were counted in young adult quail (Coturnix coturnix) after acoustic trauma at 10, 30, 60 and 90 day survival times. Following sacrifice the basilar papillae, along with the ganglia, were fixed, embedded in plastic and sectioned serially at 100 mu intervals from basal to apical tip. Hair cells and ganglion cells were counted from 3 mu thick sections at each interval. Hair cells were designated as tall or short within the area 30-70% of length from basal tip of the papilla. Both tall and short hair cells were significantly reduced in number 10 days following trauma. Tall hair cells recovered to within 96% of normal after 60-90 days. Short hair cells recovered but to a lesser extent. Ganglion cell loss did not begin until 30 days after trauma and continued without recovery 90 days after trauma. A good correlation was found for position of both types of hair cell loss and position of ganglion cell loss. These results suggest that the initial loss of hair cells, both tall and short, results in retrograde degeneration of neural fibers and ganglion cells.     

Carboplatin-radiation interaction in squamous cell carcinoma cell lines.

Chemoradiotherapy has been considered one of the most promising improvements in the treatment of advanced head and neck cancer. This article describes in vitro chemosensitivity to carboplatin in five squamous cell carcinoma cell lines established from head and neck cancers and in one vulvar squamous cell carcinoma cell line. Sensitivity to carboplatin was found to vary markedly when using the 96-well plate clonogenic assay and continuous drug exposure. The difference in carboplatin response between the most sensitive and the most resistant cell lines was fourfold. No cross-resistance was observed between inherent radiosensitivity and chemosensitivity. Effects of concomitant use of carboplatin and radiation were further investigated in the two cell lines that were found to be most sensitive to carboplatin. The drug was administered 1 hour before acute radiation doses, and an additive effect was observed in both cell lines.     

Merkel cell carcinoma

PURPOSE: To determine the natural history and treatment outcomes for patients with Merkel cell carcinoma. METHODS: Review of the literature. RESULTS: The probability of regional node involvement at presentation exceeds 50%; few patients present with distant metastases. Comprehensive treatment of the primary site and regional lymphatics with surgery or radiotherapy results in the highest likelihood of cure. The role of adjuvant chemotherapy remains investigational. CONCLUSION: The probability of regional dissemination at diagnosis is high. The optimal treatment is resection of the primary tumor and treatment of the regional lymphatics. Resection of all gross tumor should be accomplished followed by local-regional radiotherapy in most patients.     

Langerhans cell related inflammatory reaction in laryngeal squamous cell carcinoma

The major cells involved in cancer cell kill are the T lymphocytes. However, T cells need to be activated upon antigen presentation, which is mediated by the antigen presenting cells, one of which is the Langerhans cell (LC). The purpose of this study was to assess the significance of LC and inflammatory cell infiltration in the laryngeal squamous cell carcinoma (LSCC). Forty-five patients who were operated on for LSCC between 1992 and 1999 were included in the study. The clinical and histopathological features of the patients were reviewed. A semiquantitative estimation of the lymphocyte dominant inflammatory reaction within and around the tumor was performed. Anti S-100 antibodies were used for immunohistochemical detection of LCs. Horseradish peroxidase method was used. LCs were present in almost all of the specimens within and around the tumor tissue. The S-100 results did not associate with grade, T and N stages, tumor stage, laryngeal site of involvement and survival (P>0.05). The S-100 results significantly associated with inflammatory reaction in the tumor tissue (P<0.01). In conclusion, the LC related response is not important to inhibit regional metastasis by cancer cells. The LC is not a reliable tool to determine prognosis of the patients with LSCC in the clinical practice.     

Hair cell re-growth

Hair cell loss is usually a function of age, noise, ototoxic drugs and genetics. Therapeutic strategies fall into two categories: protection and regeneration. Protective methods include targeted application of growth factors and other agents to promote cell survival, and systemic application of drugs to prevent activation of programmed cell death. These strategies are related to treatments that cause predictable damage, such as the use of aminoglycoside antibiotics. The challenge of hair cell regeneration is more difficult. Instead of preventing cell loss, we must consider methods of stimulating cell division and hair cell differentiation from existing cells. We need to know much more about the molecular mechanisms that govern these processes so that we can identify potential targets for specific drugs or gene therapies. One method of approaching the issue is to combine in vitro models of developing hair cells with genomic and proteomic technologies. The benefits of hair cell re-growth may not be limited to full replacement of pre-existing cells. Surrogate hair cells may help to maintain cochlear innervation, even if they do not detect sound, and this property could be harnessed to improve the performance of cochlear implants.