成人型喉乳头状瘤中PCNA的表达和AgNORs的定量研究

探讨细胞增殖活性与成人型喉乳头状瘤生物学行为的关系。采用抗增殖细胞核抗原（ＰＣＮＡ）单克隆抗体ＰＣ１０，ＡＢＣ免疫组化法和银染技术，评价其细胞增殖活性。结果：经ＰＣＮＡ阳性反应计算出的ＰＣＮＡ增殖指数与核仁形成区嗜银蛋白（ＡｇＮＯＲｓ）计数间存在显著的相关性。经统计学分析有显著差异（Ｐ〈０．０５）。提示：ＰＣＮＡ表达和ＡｇＮＯＲｓ计数可作为喉部良、恶性病变的鉴别，检测癌前病变及观察肿瘤生物学行为较     

喉癌EGFR和PCNA表达及DNA含量的研究

研究喉鳞癌中表皮生长因子受体、增殖细胞核抗原的表达和ＤＮＡ含量,分析各指标间的相关性及与喉癌临床生物学行为和预后的关系。方法：用免疫组化ＳＰ法和图像分析仪检测正常喉粘膜、喉鳞癌的ＥＧＦＲ、ＰＣＮＡ和ＤＮＡ指数,结合临床资料进行分析。结果：喉鳞癌的ＥＧＦＲ的阳性表达率为５４．８％,与正常喉组织的差异有统计学意义,但ＥＧＦＲ的阳性表达与喉癌的病理分级和５年生丰率无显著相关,而ＰＣＮＡ阳性表达和ＤＩ则随     

Argyrophilic nucleolar organizer regions as a prognostic indicator of laryngeal carcinomas

Nucleolar organizer regions (NORs) were studied in surgically removed specimens from 76 patients with squamous cell carcinomas of the larynx using an argyrophilic (Ag) staining technique. The mean number of AgNORs per nucleus was 4.3±1.38 (SD). The mean AgNOR number for T1 or T2 disease was statistically lower than that for T3 or T4 lesions (P<0.05). The mean AgNOR number was lower in patients with N0 disease than in patients with N1 or N2 or N3 tumors (P<0.05). There was also a statistically significant difference between the mean AgNOR number for stage II and stage III disease (P<0.01), for stage III and stage IV disease (P<0.05), but not for stage I and stage II disease. According to the histological grading, there was a significant difference between the mean AgNOR number for the well-differentiated and moderately differentiated tumors (P<0.05), and for the moderately differentiated and poorly differentiated types (P<0.01). These results suggest that the number of AgNORs in the nucleus is a significant indicator of laryngeal carcinomas.     

晚期喉癌边缘DNA含量和增殖细胞核抗原表达

目的探讨晚期喉癌边缘ＤＮＡ含量和增殖细胞核抗原（ＰＣＮＡ）表达与喉癌手术预后的关系。方法通过ＰＣＮＡ表达和ＤＮＡ含量检测，对３４例晚期喉癌边缘进行观察。结果肿瘤边缘１．０、０．５、０ｃｍ处粘膜，ＰＣＮＡ指数分别为８．６２％、１７．７６％和５０．３２％。ＤＮＡ指数分别为０．９８、１．０８２和１．４３６。统计学分析，１．０、０．５ｃｍ处粘膜与０ｃｍ处粘膜差异有显著性（Ｐ＜０．０１）。结论中、重度非典型增生的上皮在分裂，增殖等生物学行为方面已与轻度非典型增生和单纯性增生的上皮细胞不同，手术切缘应在中度非典型性增生的粘膜上皮以外，即０．５ｃｍ以外。     

Nucleolar organizer regions in glottic carcinomas: Comparison of DNA cytofluorometry and clinicopathological analysis

The significance of nucleolar organizer regions (NORs) and nuclear DNA content in 73 glottic carcinomas was assessed for proliferative activity and tumor progression. NORs stained with silver colloid were counted, and nuclear DNA content was assayed by cytofluorometry. The cytofluorometric study demonstrated that the percentage of tumors with aneuploidy tended to increase as histological differentiation decreased. Survival rates of patients with diploid and aneuploid tumors were not significantly different. AgNOR staining revealed that mean AgNOR numbers rose as histological differentiation of tumors decreased. Moreover, as T and N categories and stages showed advancing malignancy, mean AgNOR numbers tended to rise. However, there was no significant difference in survival rates between tumors with low and with high AgNOR counts. These studies indicate that while AgNOR staining is better than DNA cytofluorometry for determining histological differentiation of glottic carcinoma, neither is of prognostic value at the present time.     

Nucleolar organizer regions in carcinomas of the oropharynx and hypopharynx

Summary It is supposed that the nucleolar organizer regions (NORs), genes on which ribosomal RNA is coded, may contain information on the structure of the nucleolus and on nucleolar activity which permits discrimination between normal and neoplastic cells and their proliferation activity. In the present study, the histological sections of 144 patients with carcinomas of the oropharynx and hypopharynx were studied in a retrospective approach to determine the absolute NOR counts per 100 nuclei and investigate their influence on the prognosis and extent of lymphogenic formation of metastases. An absolute NOR count of 681 per 100 nuclei was found to give reliable discrimination between stage NO or N1 carcinomas and stage N2 or N3 disease.     

A correlation of argyrophilic nucleolar organizer regions with prognoses in patients with maxillary sinus squamous cell carcinoma

Argyrophilic nucleolar organizer regions (AgNORs) were determined by a semiautomatic image analyzer in a group of 28 patients with T2 or T3 maxillary sinus squamous cell carcinoma. The group was separated into recurrent and non-recurrent groups, metastatic and nonmetastatic groups, as well as groups strongly or weakly positive for epidermal growth factor receptor (EGFR). The mean number and area of AgNOR dots per nucleus and the mean size of each dot were calculated. The mean number and area were significantly greater than those in normal cells (P < 0.001), but not in recurrent and non-recurrent groups, metastatic and non-metastatic groups and groups separated by EGFR. There was also no difference found in the mean sizes of tumor and normal cells. These results suggest that AgNOR is related to malignant transformation, but not prognosis.     

咽喉鳞癌组织中增殖细胞核抗原和核仁组成区相关嗜银蛋白的表达

目的 :研究 4 3例喉癌和 18例下咽癌术后标本中增殖细胞核抗原 (PCNA)的表达并对核仁组成区相关嗜银蛋白 (AgNOR)计数 ,分析其与临床分期、病理分化、复发、转移、生存期之间的相关性 ,评估其对预后及监测患者病情的价值。方法 :采用免疫组化法及Crocker’sAgNOR染色技术定量观察 6 1例鳞状细胞癌术后标本及9例良性喉肿瘤患者的非瘤区粘膜中PCNA的表达及AgNOR计数。分析其与喉癌及下咽癌临床分期、病理分级、复发、转移、生存期之间的相关性。结果 :PCNA阳性细胞百分率 (PCNA指数 )与AgNOR计数呈正相关 ;PC NA指数与AgNOR计数由早期到晚期 ,由高分化到低分化逐渐提高 ;两者在复发、转移及期内死亡患者的癌组织中增加 (P <0 .0 5 )。结论 :PCNA指数和AgNOR计数可作为喉及下咽鳞癌的诊断性预测指标。     

鼻内翻性乳头状瘤组织中新生血管化与细胞增殖活性的相关性研究

目的 :检测鼻内翻性乳头状瘤 (NIP)组织中CD34、增殖细胞核抗原 (PCNA)的表达 ,探讨新生血管生成与细胞增殖活性在NIP中的作用及相互关系。方法 :应用免疫组织化学SP方法检测 30例NIP、16例鼻息肉(NP)及 11例鼻腔鳞状细胞癌 (NSCC)组织中CD34、PCNA的表达 ,并在此基础上检测其微血管密度 (MVD)和PCNA阳性指数 (PCNA LI)。结果 :CD34在NP、NIP及NSCC组织中的表达分别为 (32 .4 1± 5 .6 8)、(5 3.2 2± 9.31)和 (80 .37± 10 .2 1)个 /× 2 0 0视野 ,两两比较 ,差异均有统计学意义 (均P <0 .0 1) ;PCNA LI分别为 (2 8.6 7± 9.2 9)、(35 .35±7.0 2 )和 (6 3.18± 12 .17) % ,NP与NIP比较 ,差异有统计学意义 (P <0 .0 5 ) ;NP和NIP分别与NSCC比较 ,差异亦有统计学意义 (P <0 .0 1)。PCNA LI与MVD有显著正相关关系 (r =0 .5 6 ,P <0 .0 1)。结论 :MVD、PCNA LI与细胞分化程度有很好的一致性 ,可分别作为反映NIP侵袭发展、恶变潜能的良好指标。     

Argyrophilic nucleolar organizer region counts in squamous cell carcinomas of the head and neck after irradiation and chemotherapy

After irradiation or chemotherapy of tumors of the head and neck, histological alterations might be difficult to assess. In 30 cases of squamous cell carcinomas of the pharynx, changes in argyrophilic nucleolar organizer region (AgNOR) counts were investigated after therapy. After treatment a significant decrease of AgNOR counts was detected (P < 0.001). No significant differences could be found between groups treated with radiotherapy, chemotherapy or a combination of radiotherapy and chemotherapy. These findings suggest that AgNOR counts might be useful to assess cytotoxic effects on a subcellular level. Received: 4 March 1997 / Accepted: 25 July 1997     

Clinical application of proliferating cell nuclear antigen, oncoprotein p53 and tumor front grading analysis in patients operated on for laryngeal cancer

The authors assessed proliferating cell nuclear antigen (PCNA), p-53 oncoprotein and morphologic tumor front grading (TFG) in patients with advanced squamous cell carcinoma (SCC), of the larynx and a poor prognosis and tried to find a correlation with tumor stage, the Broders grading system, local and neck lymph node metastases, as well as nodal and local recurrences. In addition, utility of the parameters investigated was evaluated in developing a prognostic factor model, using uni- and multivariate Cox regression analysis. Included in this study were 54 patients (mean age 57 years ± 8.6). PCNA-positive staining was found in all but one patient with advanced disease, while p-53 stained positively in only 24 subjects (44.4%). The PCNA index ranged from 4.6 to 59.0% (mean, 23.4 ± 11.0) and the p-53 index varied from 4.0 to 42.0% (mean, 17.2 ± 8.6). The TFG score ranged from 9 to 23 points (mean, 15.1 ± 3.2). PCNA, p-53 and TFG were found to be the markers that provided significant additional information about the biological behavior of tumor cells. The high variability of the results (PCNA, p-53) and high percentage of negatively stained cells (p-53) reduced their application in clinical use. PCNA correlated with tumor grade, G (r = 0.38; P < 0.01), but negatively with nodal (N) disease(r = –0.37; P < 0.01). The mean values of PCNA and p-53 index were higher in the subgroup with local recurrences. Our present attempt to develop a useful prognostic factor model failed. Received: 26 September 1998 / Accepted: 1 March 1999     

Apoptosis and expression of argyrophilic nucleolus organizer regions in epithelial neoplasms of the larynx

IntroductionOccurrence of apoptosis and expression of proliferative markers are powerful tools to establish a prognosis in the follow-up of cancer.ObjectiveTo evaluate the growth fraction in papillomas and laryngeal squamous cell carcinomas with three degrees of differentiation through apoptosis and the expression of nucleolus organizer regions.MethodsRetrospective study from which paraffin material was submitted to microtomy and hematoxylin–eosin and silver staining. Stained slides were used to quantify the apoptotic index and the number of nucleolus organizer regions by morphometry.ResultsApoptosis was significantly more frequent in well differentiated carcinomas and in papillomas, and a higher growth fraction of expressed nucleolus organizer regions and cells that expressed a greater than average number of nucleolus organizer regions were more frequently noted in undifferentiated carcinomas.ConclusionsThus, it was possible to verify that a high apoptotic index was associated with a lower chance of tumor differentiation in carcinomas, while a greater number of total nucleolus organizer regions, cells expressing nucleolus organizer regions above average and a higher growth fraction were associated with greater likelihood of abnormal cell proliferation and increased tumor differentiation.     

喉癌血清DBP异质性和癌组织PCNA表达检测及临床意义

应用环状免疫单扩散法及免疫组化技术对44例喉癌病人血清及32例癌组织进行血清DBP异质性和组织PCNA表达检测,发现各期喉癌病人血清中均存在DBP异常表达,并与化疗及复发有关;而与分型、淋巴结转移及病理分化无关;且血清DBP阳性喉癌组织中PCNA的表达率较高。提示它有较高的增殖活性,血清DBP异质性与喉癌的发生发展有密切关系。     

小干扰RNA对喉鳞状细胞癌裸鼠移植瘤增殖细胞核抗原及p53蛋白表达的影响

目的：应用小干扰RNA（siRNA）技术抑制裸鼠喉癌皮下移植瘤人端粒酶逆转录酶（hTERT）基因表达,探讨移植瘤中增殖细胞核抗原（PCNA）及p53蛋白表达的变化。方法：根据hTERT cDNA序列构建表达短发夹RNA（shRNA）的、靶向hTERT mRNA的真核表达质粒pshRNA,其载体质粒含荧光素报告基因。建立人喉鳞状细胞癌Hep-2细胞株裸鼠皮下接种模型,将pshRNA转染入荷瘤裸鼠瘤体内,观察肿瘤生长情况。以激光共聚焦显微镜观察质粒在瘤体内的表达;以免疫组织化学SP法检测PCNA及p53蛋白在肿瘤内的表达。结果：所有裸鼠均接种成功,5d后可见皮下肿瘤形成,14d左右肿瘤直径达5～7mm。pshRNA及空质粒载体转染入瘤体后,共聚焦显微镜下见癌组织中有绿色荧光表达。病理学检查发现：pshRNA组肿瘤生长受到抑制,细胞分裂少见,可见大量癌细胞坏死。与注射生理盐水的对照组比较,转染质粒完毕后7d抑瘤率为76．50％,P〈0．01。pshRNA治疗后瘤体内PCNA蛋白表达显著下调（P〈0．05）,p53蛋白表达显著上调（P〈0．05）。结论：靶向hTERT mRNA的shRNA可显著抑制人喉癌裸鼠移植瘤的生长,其机制可能是诱导PCNA下调,促进p53蛋白表达所致。     

A study of a new tumour marker,CYFRA 21-1, in squamous cell carcinoma of the head and neck,and comparison with squamous cell carcinoma antigen

CYFRA 21-1 (CYFRA) is a newly developed tumour marker which is useful in evaluating large cell lung carcinoma, especially the squamous cell type. The purpose of this study was to assess the clinical value of CYFRA for squamous cell carcinoma of the head and neck and compare the results with squamous cell carcinoma antigen (SCCA). Serum levels of CYFRA were measured in 168 patients with a newly diagnosed head and neck squamous carcinoma. In addition, 77 patients without evidence of neoplasm were included as controls. At the same time, SCCA was also determined. The cut-off values of CYFRA and SCCA, determined at the 95th percentile of the standard Gaussian variate of controls, were 2.48 ng/ml and 1.49 ng/ml respectively. The diagnostic sensitivity of CYFRA was superior to that of SCCA, especially for nasopharyngeal carcinoma. The sensitivity of CYFRA for nasopharyngeal carcinoma was much higher (58.3%) than that of SCCA (15.5%). However, the sensitivity of CYFRA is not satisfactory in all types of squamous carcinoma. For oral cancer, the sensitivity is only 25.6%. CYFRA is a useful serum marker for patients with certain types of head and neck squamous carcinoma, such as nasopharyngeal carcinoma and hypopharyngeal carcinoma. In addition, CYFRA may be also useful in monitoring recurrence of certain types of SCCHN, which are sometimes difficult to detect.     

Proliferating cell nuclear antigen counts as markers of cell proliferation in head and neck cancer

Proliferating cell nuclear antigen (PCNA) expression was investigated immunhistochemically in 53 squamous cell carcinomas of the head and neck. PCNA is a 36-kDa nuclear protein associated with the cell cycle. Results were compared with Ki67 counts, with this latter marker used to demonstrate proliferative compartments. Overall, the PCNA and Ki67 labelling index showed a similar distribution pattern in normal and tumor tissue. A strong correlation was found to histological differentiation. There was no significant difference between PCNA and Ki67 counts (r = 0.8), and PCNA immunostaining allowed assessment of proliferative activity in the head and neck cancers studied. The application of formalin-fixed, paraffin-embedded tumor material demonstrated the advantage of this method and showed that it is an excellent alternative to Ki67 counts.     

G3BP在喉鳞状细胞癌的表达及其临床意义

目的研究GTP酶激活蛋白SH3功能区结合蛋白（G3BP）在喉鳞癌组织中的表达情况及其与喉鳞癌生物学行为之间的关系。方法采用免疫组织化学方法检测91例喉鳞癌组织中G3BP的表达情况,并探讨他们与喉鳞癌的肿瘤组织分化程度、TNM分期、淋巴结转移和预后的关系。结果G3BP在喉鳞癌的阳性表达率为84.7％。其表达水平与肿瘤分化程度（P=0.008）、临床分期（P=0.005）、淋巴结转移（P=0.000）及肿瘤复发转移（P=0.000）均有密切关。G3BP表达水平越高,患者术后生存时间越短（P=0.000）。结论G3BP在喉鳞癌表达与肿瘤分化程度、临床分期、淋巴结转移、肿瘤复发转移情况及患者的预后有密切关系。G3BP可作为判断喉鳞癌患者预后的有效参考指标。     

A comprehensive analysis of selected diagnostic methods with respect to their usefulness in evaluating the biology of neoplastic cells in patients with laryngeal cancer

The difficult and complicated mechanism of cancer development with little knowledge about the biology of existing cancers can lead to a permanent search for new examination techniques to improve the precision of life expectancy in patients and the selection of the most efficient methods of treatment. The aim of this study was to analyze certain prognostic factors, i.e., p53, Ki67, proliferating cell nuclear antigen (PCNA), DNA ploidy and cell proliferating activity, as well as the degree of morphological differentiation and cell maturity evaluated on an ultrastructural level in patients with laryngeal cancers in connection with data obtained from follow-up examinations and the clinical course of the disease. Neoplastic tissue was taken from 120 patients with laryngeal cancers. All underwent surgical treatment, radiotherapy and combined treatment in the Department of Otolaryngology, Karol Marcinkowski University School of Medical Sciences, Poznań, Poland, and the Department of Otolaryngology – Head and Neck Surgery, Haukeland University, Bergen, Norway. Before beginning treatment all patients underwent histological verification of their neoplastic tissues. Histopathological examination proved that the majority of cases (95%) had a squamous cell carcinoma. The occurrence of changes within the lymph nodes of the neck (N) was significantly correlated with T, S, Ki67, metastases to lymph nodes, DNA ploidy, site and surgery performed. The degree of clinical progression (S) was intercorrelated with T, N, p53, Ki67, PCNA, DNA ploidy, site and laryngectomy. The occurrence of oncoprotein p53 in neoplastic cells was measured by the staining degree of their nuclei and was correlated with T, S, DNA ploidy, metastases to lymph nodes, PCNA and site. The degree of staining of neoplastic cells for the nuclear antigen Ki67 was correlated to T, N, G, S, DNA ploidy, metastases to lymph nodes and surgical treatment. The proliferative antigen PCNA in the examined population of patients was intercorrelated with T, p53, Ki67, metastases to lymph nodes and surgical treatment. The results obtained from DNA flow cytometry could be associated with N, G, p53, Ki67 and metastases to lymph nodes. On the basis of the results obtained, the techniques suggested for the morphological and biological evaluation of neoplastic cells in cancer of the larynx should include TNM classification + G + DNA + p53 + Ki67. Received: 26 September 1998 / Accepted: 13 April 1999     

p21WAF1/CIP1和抑制增殖细胞核抗原的表达与下咽癌及喉癌的临床关系

目的：探讨p21^WAF1/CIP1和抑制增殖细胞核抗原(PCNA)的表达与下咽癌、喉癌的分级、转移及预后的关系,了解p21^WAF1/CIP1在下咽癌、喉癌发生、发展中的作用以及与PCNA的相关性。方法：对45例下咽癌、25例声门上型喉癌、23例声门型喉癌标本,用Envision HIS的方法检测p21^WAF1/CIP1、PCNA的表达情况,并分析p21^WAF1/CIP1表达与下咽癌、喉癌临床病理特征、转移、复发和预后的关系。结果：p21^WAF1/CIP1阳性表达率为51.6％(48／93),PCNA的阳性表达率为97.8％(91／93);p21^WAF1/CIP1表达与下咽癌、喉癌患者的年龄、性别、肿瘤分化、远处转移及复发无显著性相关,与肿瘤的原发部位的T分期有显著性相关;p2^WAF1/CIP1的表达还与肿瘤的预后有显著性相关;而PCNA与肿瘤的临床分期及淋巴转移有显著性相关,p21^WAF1/CIP1与PCNA的表达具有相关性。结论：p21^WAF1/CIP1、PCNA是预测肿瘤浸润范围及转移的良好指标,p21^WAF1/CIP1对预后的判定具有一定的指导意义。     

COX-2、PCNA及p53在鼻咽癌中的表达及意义

目的：研究COX-2、PCNA及p53的基因产物在鼻咽癌组织中的表达及与其生物学行为和生存率之间的关系。方法:采用免疫组化SP法检测60例鼻咽癌及30例正常鼻咽组织中COX-2、PCNA 及p53的蛋白表达水平。结果:（1） 鼻咽癌组织COX-2、PCNA 及p53阳性表达率分别为90％、75％和68％ ,明显高于正常鼻咽组织;（2） 鼻咽癌组织COX-2、PCNA及p53的表达与临床分期、分化程度和淋巴结转移相关(P＜0.05);（3） COX-2、PCNA及p53的表达密切相关;（4） 鼻咽癌组织COX-2、PCNA 阳性表达和生存率呈负相关,是鼻咽癌复发的显著预后因素。结论:鼻咽癌组织中COX-2、PCNA表达水平明显上调,与鼻咽癌的发生发展及预后不良有关,免疫组化检测COX-2、PCNA表达可用以判断鼻咽癌患者的预后。