Immunohistochemical expression of EGFR and p-EGFR in oral squamous cell carcinomas

Epidermal growth factor receptor (EGFR) is a tyrosine kinase receptor of the ErbB family, which is expressed or highly expressed in a variety of solid tumors, including oral cancers. High EGFR expression has been correlated with tumor size, metastasis and survival. In recent years, EGFR has been considered a promising target for monoclonal antibody therapy. A total of 52 patients with oral squamous cell carcinoma (OSCC) were selected for EGFR and phosphorylated EGFR (p-EGFR) detection. Immunohistochemical staining was performed to evaluate EGFR and p-EGFR expression. Positive EGFR and p-EGFR staining was present in 92.3% (48/52) and 98.0% (51/52) of all cases, respectively. High EGFR and p-EGFR expression was present in 63.4% (33/52) and 69.2% (36/52) of all cases, respectively. EGFR and p-EGFR expression did not correlate with the clinical factors tumor stage, regional lymph node metastasis, or distant metastasis. However, a statistically significant correlation was identified between high EGFR expression and the pathologic factor tumor invasion. As a conclusion, the majority of OSCCs highly express EGFR and p-EGFR, indicating the importance of studying the efficacy of anticancer therapy targeting these signal factors.(Pathology Oncology Research Vol 12, No 2, 87–91)     

Zinc and copper levels in whole blood and plasma from patients with squamous cell carcinomas of head and neck.

Zinc in plasma and whole blood from 13 patients with squamous cell carcinoma of the head and neck was significantly decreased, and the copper:zinc ratio in the plasma was significantly higher than in healthy controls. The plasma zinc was significantly lower in patients who did not respond to therapy and who died within 12 months than in those who responded to therapy and had a remission within 12--15 months. In patients responding to therapy and alive after 12--15 months, zinc in the plasma and whole blood and the copper:zinc ratio became normal. These results suggest a potential screening and predicting value of zinc in the plasma and whole blood and the copper:zinc ratio in squamous cell carcinoma of the head and neck.     

Ezrin及FAK在宫颈鳞癌中的表达及意义

目的:利用量子点免疫荧光技术在宫颈鳞癌组织上进行Ezrin及FAK蛋白的检测,并评估蛋白在宫颈鳞癌中的表达及临床意义.方法:在正常宫颈(normal cervix,NC)、低度鳞状上皮内病变宫颈(LSIL)、高度鳞状上皮内病变宫颈(HSIL)和宫颈鳞癌(SCC)组织上,利用QDs标记的链霉亲和素复合物(QDs-SA)能与生物素化二抗IgG结合的特点两种方法检测Ezrin和FAK的蛋白表达.利用免疫组化SP法检测宫颈鳞癌组织中Ezrin和FAK的表达.结果:SP法观察到Ezrin、FAK蛋白分别着色于宫颈细胞的胞膜和胞浆,呈棕黄色,且在60例宫颈鳞癌细胞中阳性表达率分别为75％、80％,与以下QDs-IHC法结果相比较,差异有统计学意义;量子点免疫荧光法(QDs-IHC)观察到Ezrin、FAK蛋白分别定位于宫颈细胞的胞膜和胞浆,定位准确,并且阳性信号分别呈现绿色、橙红色的光.60例宫颈鳞癌组织中Ezrin蛋白及FAK蛋白阳性表达率分别为96.66％及95.00％,与HSIL组相比差异有统计学意义(P＜0.05).30例HSIL组织中Ezrin和FAK蛋白阳性表达率分别为83.33％和80.00％,与LSIL组相比差异有统计学意义(P＜0.05).30例LSIL组织中Ezrin和FAK蛋白阳性表达率分别为60.00％和53.33％,与正常宫颈组相比差异无统计学意义(P＞0.05).结论:Ezrin及FAK的阳性表达在宫颈鳞癌的发生发展和侵袭过程中具有一定临床意义,值得深入研究.量子点免疫荧光法(QDs-IHC)能更好地检测多个指标,更具有应用前景.     

Constitutive activating mutation of the FGFR3b in oral squamous cell carcinomas

A G to T mutation at nucleotide position 2128 in the human FGFR3b coding region resulting in a Cys for Gly substitution (G697C) in the tyrosine kinase domain was observed in 62% (44/71) of oral squamous cell carcinomas (OSCC) examined. Immunostained FGFR3b was found in the cytoplasm of prickle cells in normal epithelia, and FGFR3b was localized in the cytoplasm and nucleus in non-FGFR3b mutant OSCC. Overexpressed FGFR3b protein on plasma membranes was noted in OSCC bearing the FGFR3b mutation. Enhanced tyrosine kinase activity of G697CFGFR3b was confirmed. Our results indicate that G697C is an activating mutation causing constitutive ligand-independent FGFR3b signaling. This mutation may be involved in the progression of OSCC and thus the FGFR3b coding sequence may have diagnostic or prognostic value for OSCC.     

Expression of individual lamins in basal cell carcinomas of the skin

In this study we used a unique collection of type specific anti-lamin antibodies to study lamin expression patterns in normal human skin and in skin derived from patients with basal cell carcinomas (BCCs). Lamin expression in serial sections from frozen tissue samples was investigated by single and double indirect immunofluorescence. In normal skin, lamin A was expressed in dermal fibroblasts and in suprabasal epithelial cells but was absent from all basal epithelial cells. Lamin C was expressed in dermal fibroblasts, suprabasal epithelial cells and a majority of basal epithelial cells. However, lamin C was not expressed in quiescent basal epithelial cells. Lamin B1 was expressed in all epithelial cells but was not expressed in dermal fibroblasts. Finally, lamin B2 was expressed in all epithelial cells but was not expressed in dermal fibroblasts. Finally, lamin B2 was expressed in all cell types in normal skin. Lamin expression was also investigated in a collection of 16 BCCs taken from a variety of body sites. Based upon patterns of lamin expression the BCCs were classified into four groups: A-negative (10/16 tumours), C-negative (5/16 tumours), A/C-negative (1/16 tumours) and A/B2-negative (1/16 tumours). Lamin expression was also compared to cell proliferation index by staining serial sections with the proliferation marker Ki67. 9/10 of the lamin A negative tumours were highly proliferative, whereas 4/5 of the lamin C negative tumours were slow growing. Thus as a general rule absence of lamin A was correlated with rapid growth within the tumour, while absence of lamin C was correlated with slow growth within the tumour. Our data supports the hypothesis that lamin A has a negative influence on cell proliferation and its down regulation may be a requisite of tumour progression.     

埋植式动脉泵化疗与口腔鳞癌的综合治疗

目的:探讨埋植式动脉泵在口腔鳞癌化疗中的效果及在口腔鳞癌综合治疗中的应用价值。方法:采用ZW埋植式动脉泵经颈外动脉插管治疗口腔鳞癌29例,观查化疗效果及其与手术、放疗综合应用的可行性。结果:埋植式动脉泵给药治疗口腔鳞癌的有效率87.5%,未出现明显毒副作用。结论:埋植式动脉泵是口腔鳞癌化疗给药方法的较好选择,可作为多药物、多周期单一化疗;术前诱导及术后辅助化疗;化疗与放疗联合应用,多手段综合治疗中化疗最佳途径,可在临床广泛应用。     

埋植式动脉泵化疗与口腔鳞癌的综合治疗

目的：探讨埋植式动脉泵在口腔鳞癌化疗中的效果及在口腔鳞癌综合治疗中的应用价值。方法：采用ZW埋植式动脉泵经颈外动脉插管治疗口腔鳞癌29例,观查化疗效果及其与手术、放疗综合应用的可行性。结果：埋植式动脉泵给药治疗口腔鳞癌的有效率87.5%,未出现明显毒副作用。结论：埋植式动脉泵是口腔鳞癌化疗给药方法的较好选择,可作为多药物、多周期单一化疗;术前诱导及术后辅助化疗;化疗与放疗联合应用,多手段综合治疗中化疗最佳途径,可在临床广泛应用。     

Second primary cancers in patients with squamous cell carcinoma of the skin

The occurrence of second primary cancers was explored in patients with squamous cell cancer of the skin (SCC). The excess incidence subsequent to SCC was mainly in cancers related to sunlight and smoking, and in lymphoproliferative malignancies, it was largest (10-fold) in salivary gland cancer.     

凋亡相关基因蛋白bcl—xl和bak在肺鳞癌中的表达及意义

目的：探讨肺鳞状细胞癌中凋亡调控因子bcl—xl和bak蛋白的表达及其意义。方法：采用免疫组织化学技术检测50例肺鳞状细胞癌及20例正常肺组织中bcl—xl和bak蛋白的表达水平。结果：bcl—xl蛋白在肺鳞状细胞癌中表达水平高于正常肺组织（P〈0．05）。bak表达水平低于正常肺组织（P〈0．05）。bcl—xl和bak蛋白的表达水平在不同分化程度的肺癌之间差异有显著意义（P〈0．05）。结论：肺鳞状细胞癌的发生、发展与bcl—xl和bak蛋白的异常表达有关,可能是细胞凋亡失衡的结果。     

Effects of an anti-angiogenic agent, TNP-470, on the growth of oral squamous cell carcinomas

Anti-tumor effects of an anti-angiogenic agent, TNP-470, on the growth of oral squamous cell carcinomas (SCCs) were evaluated in vivo and in vitro. The growth of an oral SCC cell line, HSC-2, inoculated subcutaneously in severe combined immuno-deficiency (SCID) mice was inhibited in a dose-dependent manner by the treatment with this agent. A reduction of microvessels surrounding tumor tissues treated with TNP-470 was observed by immunohistochemistry. Significant side-effects were not observed except for weight loss during the period of treatment with high dose (50 mg/kg) of TNP-470. The direct effects of TNP-470 on oral SCC cell lines were also evaluated in culture. The growth of all eight SCC cell lines tested was inhibited by TNP-470, but the sensitivity of the oral SCC cell lines to TNP-470 was about 1700 times less than that of endothelial cells. These results suggest that TNP-470 inhibits the growth of oral SCC by anti-angiogenic activities and that it may be effective as a new therapy of oral cancer.     

Thrombospondin-1 expression in oral squamous cell carcinomas: correlations with tumor vascularity, clinicopathological features and survival

Thrombospondin-1 (TSP-1) is a 450 kd glycoprotein synthesized and incorporated into the extracellular matrix by numerous cell types and reported to suppress tumor growth and progression by its inhibition of angiogenesis. In order to clarify the biological role of TSP-1 and determine its clinicopathological significance in oral squamous cell carcinomas (SCCs), we identified TSP-1 protein expression in 54 oral SCCs by immunohistochemistry and correlated it with microvessel density (MVD), clinicopathological features and patient's survival. Thirty-two out of 54 carcinomas (59%) were identified as having a low level of TSP-1 expression (TSP-1-L), and 22/54 (41%) carcinomas identified as having a high level of TSP-1 expression (TSP-1-H). The MVD counts (mean±S.D.=9.0±4.9) in TSP-1-H tumors was significantly lower than that (mean±S.D.=12.7±4.4) in TSP-1-H tumors (P=0.0065). The level of TSP-1 expression was not correlated with T category and histologic grade, but inversely correlated with the pattern of tumor invasion (P=0.0136) and with lymph nodal status (P=0.0119). Furthermore, Kaplan–Meier analysis showed that the 5-year survival rate of TSP-1-H group was significantly higher than that of TSP-l-L group. Our results suggested that TSP-1 expression exerts an inhibitory effect on tumor vascularity, and that it has value in assessment of aggressiveness and prognosis of oral SCCs.     

Prognostic impact of sterilized lymph nodes in esophageal squamous cell carcinomas after neoadjuvant chemoradiotherapy

BackgroundThe prognostic importance of sterilized lymph nodes (SLN) remains unclear in patients with esophageal squamous cell carcinomas (ESCC) treated by neoadjuvant chemoradiotherapy (nCRT). This study aimed to determine whether SLN predicted disease-free survival (DFS) in ESCC.MethodsWe enrolled 246 eligible patients who were divided into SLN (+) and SLN (-) group according to the presence or absence of fibrosis, necrosis, calcifications and/or foreign body giant cell reactions in the negative lymph nodes specimens. The prognostic value of SLN was determined using univariate and multivariate analyses. The prognostic strength of counting SLN as positive lymph nodes was evaluated using the difference of Akaike information criterion (ΔAIC).ResultsA total of 61 SLN were identified in 38 (15.4%) patients. There was no significant difference in baseline characteristics between SLN (+) and SLN (-) group. The most frequently detected SLN in the thoracic cavity and abdominal cavity were those along bilateral recurrent laryngeal nerve (21/38,55.3%) and left gastric artery (13/24,60.9%), respectively. The univariate and multivariate analyses showed SLN was an independent prognostic factor for worse DFS in the whole cohort (HR = 2.05, 95%CI = 1.08–3.90, P = 0.029). The SLN (+) group additionally correlated with worse 5-year DFS than SLN (-) group in the ypT0, ypN0 and pCR subgroups. Counting SLN as positive lymph nodes showed better prognostic strength than ignoring them.ConclusionSLN was of prognostic significance for worse DFS in patients with ESCC, particularly in patients with good response to nCRT.     

EGFR protein overexpression and gene amplification in squamous cell carcinomas of the esophagus

Overexpression of epidermal growth factor receptor (EGFR) is observed in many cancers, sometimes accompanied by gene amplification. Recently, several clinical therapies targeting EGFR were developed, but the eligibility criteria for these therapies is not fully established. To develop such eligibility criteria for esophageal squamous cell carcinoma (ESCC), we sought to clarify: (i) the exact frequency of EGFR overexpression, (ii) the relationship between protein overexpression and gene amplification, (iii) the relationship between gene amplification and specific gene mutations and (iv) the correlation between the status of EGFR and clinical or pathological features. Immunohistochemistry revealed that EGFR protein is overexpressed in 53 (50%) of the 106 ESCC examined. Fluorescence in situ hybridization (FISH) indicated clear EGFR gene amplification in 15 of the 53 tumors, somewhat higher EGFR copy in 32 cases, and no increase in 6 cases. Gene amplification was significantly associated with high level overexpression. Direct sequencing of exons 19 and 21 of EGFR revealed no mutations in 15 tumors exhibiting gene amplification, and no mutations in 25 tumors not exhibiting gene amplification. Overexpression of EGFR was significantly correlated with depth of invasion of the tumor.In conclusion, anti-EGFR therapies may be appropriate for patients with ESCC. We assume that combined analyses by immunohistochemistry/FISH would clarify aberrations in protein and gene function, and could help to identify those patients who may benefit from anti-EGFR therapy.     

长链非编码RNA H19对外阴鳞癌细胞A431增殖、迁移及凋亡的影响

目的:探讨长链非编码RNA H19在外阴鳞癌细胞系A431、人正常表皮细胞Hacat中的表达,及沉默H19表达后细胞增殖、迁移及凋亡的变化.方法:采用qRT-PCR方法检测A431细胞中H19的表达水平;用siRNA瞬时转染A431细胞,采用CCK8法、Transwell小室及Annexin V-FITC/PI双标记实验分别检测H19对A431细胞增殖、迁移及凋亡的影响.结果:H19在外阴鳞癌细胞系A431中的表达明显高于人正常表皮细胞Hacat,A431细胞中H19的表达下调能够抑制A431细胞的增殖及迁移能力,并且促进凋亡.结论:H19-siR-NA下调H19的表达,抑制A431细胞的增殖、迁移能力,促进凋亡,提示H19在外阴鳞癌中可能发挥促癌作用,其异常表达可能与外阴鳞癌的发生发展密切相关.     

口咽鳞癌中HPV感染与血管生成的关系

目的:研究口咽鳞癌(oropharyngeal squamous cell carcinomas,OSCC)中人乳头瘤病毒(human papillomavirus,HPV)感染与血管内皮生长因子(vascular endothelial growth factor,VEGF)、CD31标记的微血管密度(microvessel density,MVD)表达的关系,探讨它们在口咽鳞癌发生中的相互关系.方法:采用多重实时荧光定量聚合酶链反应(multiplex real-time polymerase chain reaction,MT-PCR)检测60例口咽鳞癌HPV DNA及型别,通过HPV DNA分析进行HPV感染状况的测定;同时采用半定量免疫组化检测HPV(+)组和HPV(-)组中VEGF、CD31(MVD标志)蛋白表达情况.结果:口咽鳞癌中HPV感染率36.7％(22/60),HPV-16型占所有感染者的比例86.4％ (19/22),明显高于其他型别;VEGF高度表达者出现MVD＞ 15个/高倍视野的几率较VEGF低度表达者高(P =0.001);MVD＞ 15个/高倍视野的肿瘤为低分化的可能性较高(P =0.004);VEGF和MVD表达与患者年龄、性别、TNM分期、HPV感染状况之间无相关性.结论:HPV感染与口咽鳞癌的发生存在相关性.口咽鳞癌中HPV感染状况和VEGF之间无相关性.     

口腔鳞癌中心体扩增与p53 STK15蛋白表达相关性

  目的  分析STK15及p53蛋白表达与口腔鳞状细胞癌（OSCC）中心体扩增相关性,探讨OSCC中心体扩增可能分子调控机制。  方法  8例正常口腔黏膜及43例OSCC石蜡包埋组织,采用间接免疫荧光双重染色了解OSCC中心体数目扩增状况,采用免疫组织化学方法检测相应组织p53、STK15蛋白表达情况,并分析其与中心体扩增相关性。  结果  中心体扩增可见于76.74%（33/43）OSCC中。OSCC中STK15阳性率为67.44%（29/43）,STK15阳性率在p53阳性组高于p53阴性组（P < 0.05）。OSCC中心体扩增发生率在STK15/p53阳性共表达组高于STK15/ p53阴性组（P < 0.05）,Spearman相关分析显示STK15/ p53阳性共表达与OSCC中心体扩增之间存在相关关系（r=0.356,P=0.019）。  结论  中心体扩增是OSCC常见异常现象,p53/STK15转激活-非依赖通路可能参与中心体异常的形成,与之共同参与OSCC的发生。      

中期因子在喉癌中的表达及其临床意义

目的 检测中期因子(Midkine,MK)在喉癌组织中的表达,探讨MK高表达与喉癌颈淋巴结转移及预后的关系。方法 应用免疫组化法检测喉癌组织中MK的表达水平及微血管密度(MVD)。结果 14例喉正常黏膜和14例乳头状瘤组织中未见MK蛋白染色或仅见微弱染色(-)(2例乳头状瘤),表达率为0;而在75例喉癌中35例弱阳性(+),21例强阳性(++),表达率为74.7%。肿瘤细胞的MK与生存时间有关(P=0.001),MK表达从阴性(-)到阳性(+、++)生存期较原来下降5.63倍,应用多因素分析结果表明淋巴结的有无与复发有关,有淋巴结转移的复发是无淋巴结转移的5.488倍。结论 MK在喉癌组织中的高表达与颈淋巴结转移密切相关,可以作为喉癌诊断、预后的判定指标。