程序性死亡受体1及其配体调控变应性鼻炎免疫偏移的实验研究

目的探讨程序性死亡受体1(PD-1)及其配体PD-L1、PD-L2在变应性鼻炎(AR)中的表达及其对AR免疫偏移的调控作用。方法收取2018年5月至2019年5月,因鼻腔高反应症状于武汉大学人民医院耳鼻咽喉头颈外科门诊就诊,或因鼻中隔偏曲住院行鼻中隔矫正手术的82例患者,男42例,女40例,年龄14~38岁。收集相应患者的临床资料、血液、下鼻甲黏膜标本。根据临床表现、皮肤点刺试验和血清特异性变应原筛查特异性IgE(sIgE)检测结果,将患者分为AR组和对照组。免疫组织化学法检测鼻黏膜中PD-1及配体的表达情况。流式细胞术检测外周血PD-1^+CD4^+T细胞、PD-L1^+髓样树突状细胞(mDCs)、PD-L2^+mDCs及Th2细胞比例。酶联免疫吸附(ELISA)法检测血清中总IgE、PD-1及配体分泌型(sPD-1、sPD-L1、sPD-L2)的表达水平。正态分布或经Ln转换后呈正态分布的计量资料组间比较采用t检验,相关性分析采用Spearman或Pearson相关,设P<0.05为差异有统计学意义。结果AR组鼻黏膜中免疫细胞表面PD-1及其配体的表达明显高于对照组。AR组外周血中PD-1^+CD4^+T细胞、PD-L1^+mDCs、Th2细胞比例明显高于对照组[(15.24±6.45)%比(8.71±5.33)%,(8.79±2.01)%比(5.74±2.90)%,(7.89±1.95)%比(2.52±1.34)%,P值均<0.05],两组PD-L2^+mDCs比例无明显差异。PD-1^+CD4^+T细胞比例与AR病情总体视觉模拟量表(VAS)评分、血清总IgE、sIgE浓度呈正相关(r值分别为0.501、0.541、0.608,P值均<0.05),PD-L1^+mDCs比例与AR病情总体VAS评分、sIgE浓度呈正相关(r值分别为0.604、0.563,P值均<0.05),Th2细胞比例与PD-L1^+mDCs、PD-1^+CD4^+T细胞比例呈正相关(r值分别为0.538、0.623,P值均<0.05)。AR组sPD-1、sPD-L1及总IgE表达水平较对照组明显升高[(4.40±1.01)pg/ml比(3.79±1.21)pg/ml、(3.88±0.25)pg/ml比(3.57±0.23)pg/ml、(6.34±1.38)ng/ml比(4.89±1.10)ng/ml,P值均<0.05],sPD-L2表达水平无明显差异。相关性分析发现sPD-L1与总IgE、sIgE浓度呈正相关(r值分别为0.32、0.45,P值均<0.05)。结论AR患者外周血及鼻黏膜组织中免疫细胞表面均高表达PD-1及PD-L1,AR患者外周血中sPD-1及sPD-L1表达水平升高,PD-1/PD-L1信号通路可通过诱导Th2型免疫反应促进AR炎性反应。     

Glut3 expression in biopsy specimens of laryngeal carcinoma is associated with poor survival

OBJECTIVES/HYPOTHESIS: The aim of the study was to determine the clinical significance of the expression of Glut1 and Glut3 proteins in biopsy specimens of squamous cell carcinoma (SCC) of the larynx. STUDY DESIGN: A retrospective study. METHODS: Using immunohistochemistry, we immunostained sections of formalin-fixed, paraffin-embedded tissues from 48 biopsies of invasive SCC of the larynx for Glut1 and Glut3. The percentages of positive cells were recorded, then correlated with overall patient survival using the Kaplan-Meier method and the Breslow-Gehan-Wilcoxon test for statistical significance. RESULTS: All cases were positive for Glut1, and Glut1 expression was not associated with survival difference at any cut-off value. Eighteen (38%) of the cases were Glut3-negative and 30 (62%) were Glut3-positive. Glut3-positive cases were associated with poorer survival than Glut3-negative cases (P =.0336). No significant difference was found between Glut3-negative and Glut3-positive groups in respect to sex, tumor site (glottic vs. supraglottic), nodal or distant metastasis, or treatment modality. However, there were significantly more poorly differentiated tumors in the Glut3-positive group than in the Glut3-negative group (27% vs. 0%, respectively; P =.0182, Fisher's Exact Test). After poorly differentiated tumors were excluded from the survival analysis, Glut3 immunoreactivity remained a significant marker of poor prognosis (P =.0385). CONCLUSION: Immunohistochemical detection of Glut3 in biopsy specimens of SCC of the larynx is a marker of poorer prognosis.     

MIB-1 and involucrin expression in laryngeal squamous carcinoma: the relationship to host and tumour factors and survival

MIB-1 is an antibody which attaches to the Ki67 antigen expressed by proliferating cells. MIB-1 immunoreactivity may be used to quantify the proliferative component of a tumour. Involucrin is a protein expressed by mature keratinocytes and may be used as a marker of differentiation. The present paper studies the expression of these two markers in a group of patients with squamous carcinoma of the larynx. Tumour cell kinetics were studied in 49 patients with squamous cell carcinoma of the larynx using antibodies to `Ki67' and involucrin. The median potential follow-up for the group was 8.1 years with a minimum follow-up of 5 years. The median MIB-1 index was 32%. The median involucrin index was 56%. Fifteen patients had no or only slight involucrin staining whereas 34 stained intensely for this protein. Involucrin expression was found to be associated with histological grade with those patients expressing involucrin tending to have well differentiated tumours and those not expressing this parameter tending to have poorly differentiated tumours ( P = 0.045). There were no other associations between host and tumour factors and the various biological parameters. Survival analysis demonstrated that patients with an involucrin count above the median value had a better 5-year survival than those below the median (89% and 56% respectively) ( P < 0.05). In addition, patients with no (or poor) involucrin expression had an increased risk of developing a recurrence at the primary site ( P < 0.05). Involucrin appears to be a promising marker of tumour differentiation and survival in squamous carcinoma of the larynx.     

Loss of p21 expression is associated with p53 mutations and increased cell proliferation and p27 expression is associated with apoptosis in maxillary sinus squamous cell carcinoma

Conclusions. Loss of p21 expression dependent on the p53 mutation may be associated with higher tumor cell proliferation, and low p27 expression may be associated with decreased spontaneous apoptosis, resulting in poorer prognosis in patients with maxillary sinus squamous cell carcinoma (SCC). Objective. We have previously reported that p53 mutations and decreased spontaneous apoptosis were associated with poor prognosis in maxillary sinus SCC. However, whether p21 and p27 expression and cell proliferation correlate with either p53 status, spontaneous apoptosis or prognosis in maxillary sinus SCC has not been evaluated. Material and methods. Seventy patients with maxillary sinus SCC were analyzed. Tumor biopsy specimens were examined for p21 and p27 expression using an immunohistological method. The percentage of proliferating cells labeled by anti-Ki-67 mAb was expressed as the Ki-67 index (KI). Results. Loss of p21 expression correlated with p53 mutations (p=0.0072). The KIs in patients without p21 expression and with p53 mutations were significantly higher than those in patients with p21 expression (p=0.0119) and those without p53 mutations (p=0.0048). Patients with p27 expression showed a significantly higher apoptotic index than those without (p=0.0012). Kaplan–Meier analysis showed that p21 expression was closely associated with prolonged disease-free survival in the group with a normal p53 status (p=0.0472). Multivariate analysis identified high KI as an independent prognostic marker (p=0.047).     

Endoglin expression is associated with poor oncologic outcome in oral and oropharyngeal carcinoma

Conclusions. The present preliminary results suggest that endoglin (CD105)-assessed micro-vessel density (MVD) in primary oral and oropharyngeal squamous cell carcinomas (SCCs) may identify patients at risk of disease recurrence or poor oncological outcome after treatment. Objectives. MVD is an independent prognostic indicator in several human malignancies. Endoglin antibodies have shown a greater specificity for tumour vasculature in comparison with pan-endothelial markers. The present explorative study evaluated endoglin expression and its prognostic role in oral and oropharyngeal SCCs. Patients and methods. The study considered 13 consecutive cases of oral and oropharyngeal SCC with lymph node metastases (pN?+?) and 13 consecutive pN0 cases. CD105-assessed MVD was calculated at 400× magnification. Results. The mean MVDs were 3.6 and 3.1 in pN+ and pN0 groups, respectively (p>0.05). The mean CD105-assessed MVDs were 4.7 in the group with locoregional recurrence and 2.9 in the group without locoregional recurrence or post-treatment diagnosis of distant metastasis (p=0.01). The mean CD105-assessed MVD in primary oral and oropharyngeal SCCs with poor oncological outcome (recurrence of disease or occurrence of distant metastasis) was 4.3. The mean MVD in primary oral and oropharyngeal SCCs with good outcome was 2.9. Statistical analysis showed a significant difference between CD105-assessed MVD in poor and good outcome groups (p=0.02).     

Nuclear expression of onco-suppressors nm23-H1 and maspin are associated with lower recurrence rate in laryngeal carcinoma

Purpose

The main aim of the study was to preliminarily investigate the possibly related role of nuclear onco-suppressors maspin and nm23-H1, a metastasis suppressor, in laryngeal squamous cell carcinoma (LSCC).

Dickkopf-1和GATA-6在喉癌中的表达及意义

目的：研究Dickkopf-1和GATA-6蛋白在喉癌组织中的表达,探讨二者在喉癌发生、发展中的作用及二者的相关性。方法：应用免疫组织化学方法（SP法）检测48例喉癌组织及48例相应癌旁组织、20例非喉癌患者喉部正常黏膜组织中Dickkopf-1和GATA-6蛋白的表达情况。结果：①Dickkopf-1蛋白在喉癌组织中的阳性表达率明显低于癌旁组织和正常喉部黏膜组织（P〈0.05）;②GATA-6蛋白在喉癌组织中的阳性表达率明显高于癌旁组织和正常喉部黏膜组织（P〈0.05）;③Dickkopf-1和GATA-6蛋白在喉癌组织中的表达与喉癌淋巴结转移、临床分期、病理分级有相关性（P〈0.05）;④喉癌组织中Dickkopf-1蛋白表达和GATA-6蛋白表达呈负相关（r=-0.490,P〈0.01）。结论：Dickkopf-1和GATA-6蛋白的表达与喉癌的发生、发展有关。     

Association between high initial tissue levels of cyclin d1 and recurrence of nasopharyngeal carcinoma

OBJECTIVES/HYPOTHESIS: Cyclin D1 expression and the rate of apoptosis have been reported to serve as important prognostic indicators in human cancers. The purpose of the present study was to determine the prognostic significance of both initial cyclin D1 expression and the apoptotic index in nasopharyngeal carcinoma. STUDY DESIGN: Cohort study. METHODS: Cyclin D1 protein levels and apoptosis in tumors and their corresponding adjacent, histologically normal tissues were determined at the time of initial diagnosis using immunohistochemical staining, Western blot analysis, and in situ end labeling, respectively, in 64 patients with T1-T4/N0-N2, poorly differentiated squamous cell carcinoma of the nasopharynx. All cases were treated by routine radiation therapy with a total median dose of 70 Gy and followed up for 10 years. RESULTS: High levels of cyclin D1 were found in 35 of 64 tumor specimens (54.7%); no cyclin D1--positive cells were determinable in normal epithelium of the nasopharynx. Rates of early local recurrence (within 5 y) were significantly higher (P <.01) for patients with high levels of cyclin D1 before radiation therapy (24 of 35 patients [68.6%]) as compared with patients with low or no expression (3 of 29 [10.3%]). Furthermore, patients bearing high levels of cyclin D1 had a poorer prognosis concerning 10-year survival than the others (P <.001), whereas overexpression of cyclin D1 did not correlate with the initial TMN classification (P >.05). According to the rate of spontaneous apoptosis in tumors below or above the median, patients were divided into two groups. There was no statistically significant difference for the overall survival between the two groups (P >.05). CONCLUSIONS: The present study demonstrates that cyclin D1 can be used as an indicator of recurrence and subsequent prognosis in nasopharyngeal carcinoma after radiation therapy. At the same time, the apoptotic state before radiation therapy is of no value in predicting the prognosis of patients with nasopharyngeal carcinoma.     

Overexpression of transketolase TKTL1 is associated with shorter survival in laryngeal squamous cell carcinomas

Tumorigenesis involves energy production by aerobic glycolysis (“Warburg effect”) in malignant tumors. One of the key enzymes is transketolase. Transketolase, transketolase-like-1 (TKTL1), and transketolase-like-2 are known. Antibodies against TKTL1 exist for immunohistochemical investigations. This study investigated the influence of TKTL1 on survival and metastasizing in 40 laryngeal squamous cell carcinomas (SCCs, T2–T4, 27 metastasized). Staining was assessed by an immunoreactive score (IRS) with values from 0 to 12 in primaries and their nodal metastases. The highest IRS was 8. Normal epithelium did not show an expression. Three SCCs were negative. Advanced SCCs had a higher IRS than lower stages. An IRS > 4 was associated with a shorter disease specific survival, independent on the tumor stage in the multivariate analysis. Significant differences between metastasized and non-metastasized SCCs were absent, but poorly differentiated SCCs had a higher IRS in their metastases than moderate differentiated SCCs. TKTL1 overexpression is associated with a more aggressive behavior and shorter survival of laryngeal SCCs. These observations could lead to additional therapeutic options targeting a blocking of the enzyme activity.     

p53 and cyclin D1 as prognostic factors in squamous cell carcinoma of the larynx

OBJECTIVES/HYPOTHESIS: Proteins p53 and cyclin D1 play a crucial role in cell cycle control. Protein p53 mutations are one of the most common genetic alterations in human cancer, and cyclin D1 gene amplification has been found to be associated with poor prognosis in different types of tumors. Functional alterations of these proteins may play an important role both in the carcinogenesis of squamous carcinomas of the head and neck and in the clinical evolution of these tumors. The objective of the present study was to evaluate whether the presence of p53 and/or cyclin D1 proteins (detected by immunohistochemical analysis) could serve as relevant variables for the assessment of the prognosis of laryngeal squamous cell carcinoma. STUDY DESIGN: Retrospective multicentric study. METHODS: Paraffin-embedded biopsy specimens from 62 human epidermoid laryngeal carcinomas were randomly selected. The expression of p53 and cyclin D1 was examined by means of immunohistochemical analysis with a view to evaluating whether there is a correlation between the aberrant expression of these proteins and disease prognosis. RESULTS: In the sample, the presence of immunohistochemically detectable p53 is associated with shorter survival and disease-free intervals, as shown in Kaplan-Meier survival curve analysis. Indeed, multivariate statistical analysis revealed that the accumulation of p53 is an independent prognostic factor, which is associated with shorter survival. This association was not evident in the case of cyclin D1. CONCLUSION: A more precise prognosis of patients with laryngeal epidermoid carcinomas could be achieved by taking into account the presence of p53 (as assayed by immunohistochemical analysis) in biopsy tissue     

A review of clinical and histological parameters associated with contralateral neck metastases in oral squamous cell carcinoma

Oral squamous cell carcinoma(OSCC)bas a high incidence of cervical micrometastases and sometimes metastasizes contralaterally because of the rich lymphatic intercommunications relative to submucosal plexus of oral cavity that freely communicate across the midline,and it can facilitate the spread of neoplastic cells to any area of the neck consequently.Clinical and histopathologic factors continue to provide predictive information to contralateral neck metastases(CLNM)in OSCC,which determine prophylactic and adjuvant treatments for an individual patient.This review describes the predictive value of clinical-histopathologic factors,which relate to primary tumor and cervical lymph nodes,and surgical dissection and adjuvant treatments.In addition,the indications for elective contralateral neck dissection and adjuvant radiotherapy(aRT)and strategies for follow-up are offered,which is strongly focused by clinicians to prevent later CLNM and poor prognosis subsequently.     

BUB1在喉鳞状细胞癌中的表达及临床意义

目的：探讨喉鳞状细胞癌（LSCC）组织中BUB1蛋白的表达及临床意义。方法：采用免疫组织化学EliVision二步法检测55例LSCC组织和30例癌旁正常黏膜组织中BUB1蛋白的表达,并分析其与LSCC临床病理特征的关系。结果：BUB1蛋白在LSCC组织中的阳性表达率为50．9％（28／55）,低于癌旁正常黏膜组织83．3%（25／30）,两者比较差异有统计学意义（P〈0．01）;BUBl蛋白的表达与LSCC分化程度及淋巴结转移有相关性（均P〈0．05）,而与患者性别、年龄、吸烟、肿瘤原发部位、T分期及临床分期无相关性（均P〉0．05）。结论：BUBl蛋白的低表达与LSCC的发生、发展有关,BUB1可能作为预测LSCC转移及预后的生物学指标之一。     

鼻咽癌中环氧化酶-2和潜伏膜蛋白-1的表达和相互关系的初步研究

目的：探讨鼻咽癌组织中的环氧化酶2（COX-2）、潜伏膜蛋白-1（LMP1）的表达情况,同时分析了两者之间的关系。方法：53例鼻咽癌活检标本,8例鼻咽部慢性炎症常规石蜡切片标本作为对照组,用SP免疫组织化学法检测COX-2、LMP1的表达,采用sPsS12．0统计软件对实验数据进行统计。结果：在鼻咽癌组织中,COX-2阳性表达率为71．70％（38／5a）;LMPI阳性表达率为66．04％（35／53）。COX2、LMP1的表达在颈部淋巴结转移组中均明显高于无颈部淋巴结转移组（P％O．05）,但在年龄、性别、病理类型、临床分期的分组中均差异无统计学意义（均P〉0．05）。在鼻咽癌组织中,LMP1和COX-2的表达存在正相关（7—0．797,P〈0．01）。结论：COX-2、LMP1在鼻咽癌组织中均有表达。COX2、LMP1在鼻咽癌中的表达和鼻咽癌颈淋巴结转移存在显著相关性,但和年龄、性别、病理类型、临床分期无相关性。COX-2和LMP1在鼻咽癌组织中的表达呈正相关。LMP1可能通过多途径上调COX-2的表达,从而促进鼻咽癌颈部淋巴结转移。     

生长抑制基因1在喉鳞状细胞癌中的突变检测及与p53蛋白表达关系的研究

目的:研究喉鳞状细胞癌组织中生长抑制基因1(ING1)基因的突变情况,并探讨p33ING1b与p53蛋白表达的关系。方法:抽提33例石蜡固定喉鳞状细胞癌组织DNA,PCR扩增ING1第2外显子序列,测序检测突变情况;免疫组织化学法检测喉鳞状细胞癌p33ING1b蛋白与p53蛋白的表达水平,并分析两者之间的关系。结果:喉鳞状细胞癌组织中ING1基因无突变现象,但2例组织中出现编码区第170个密码子核苷酸多态性位点,由GGG变为AGG;免疫组织化学检测结果显示p33ING1b、p53蛋白表达与喉鳞状细胞癌临床病理参数之间无相关性。但配对χ2分析显示p33ING1b、p53蛋白表达有明显差异,在p53阳性的病例中,p33倾向于阴性表达。结论:喉鳞状细胞癌中ING1基因的突变较少见,在正常生理环境中,p33ING1b可能与野生型p53蛋白协同发生作用。     

SMARCB1(INI-1)缺失性鼻腔鼻窦癌的诊疗进展

SMARCB1(INI-1)缺失性鼻腔鼻窦癌(SDSC)是一类罕见的鼻腔鼻窦恶性肿瘤,具有高度侵袭性。免疫组化染色检测细胞核中INI-1蛋白表达缺失是诊断SDSC最具价值的方法,由于临床表现的非特异性,大多数患者被确诊为该疾病时已处于晚期,极大程度地降低了患者的生存率及生存质量。手术完整切除肿物、术后辅以放/化疗的综合治疗模式是目前临床工作中的主要治疗手段,但仍有较高的复发率及死亡率。而新辅助治疗以及靶向治疗对于该疾病的有效性仍处于临床试验阶段。因此,早期诊断及探索最佳的治疗策略对患者至关重要。本文就SDSC的诊断及治疗进展进行综述,以助于提高对此类罕见肿瘤类型的临床认知。     

下咽鳞状细胞癌组织claudin-1的表达及与微淋巴管生成的关系

目的 通过检测97例下咽鳞状细胞癌患者癌组织中claudin-1与D2-40蛋白的表达,分析claudin-1与微淋巴管密度(MLVD)之间的相互关系,探讨它们与肿瘤临床分期、病理学分级及患者临床预后之间的相关性,探索下咽鳞状细胞癌淋巴结转移的机制.方法 应用免疫组化法检测97例下咽鳞状细胞癌及90例癌旁组织标本中cl...