Nucleolar organizer region count and subjective AgNOR pattern assessment (SAPA) score in skin tumors.

OBJECTIVE: To study the argyrophilic nucleolar organizer region (AgNOR) count and subjective AgNOR pattern assessment (SAPA) score in cytologic and histologic specimens of various skin tumors. STUDY DESIGN: The study group consisted of 37 patients (14 benign and 23 malignant) of various skin tumors. In all cases, cytology by fine needle aspiration cytology (FNAC), and histological specimens were studied by conventional staining and silver staining for AgNOR. RESULTS: The mean count in benign tumors in cytologic specimens was 2.08 +/- 0.01, compared with 5.50 +/- 1.12 in malignant tumors (P<0.001). In histologic specimens, mean count was 2.13 +/- 0.51 in benign, compared with 5.38 +/- 1.10 in malignant tumors (P<0.001). The SAPA score in benign tumors (P<0.001) in cytologic specimens, was 6.07 +/- 0.83, compared with 10.65 +/- 1.27 in malignant tumors, and in histology, it was 6.07 +/- 0.87 in benign, compared with 10.83 +/- 1.15 in malignant tumors (P < 0.001). Melanoma showed the higher AgNOR count compared with squamous cell carcinoma and basal cell carcinoma. The parameters were statistically significant between the grade of tumor in squamous cell carcinoma and the positivity of lymph nodes as demonstrated by SAPA score. No correlation was found between the clinical stage and Clark level of melanoma. Although, AgNOR count and SAPA score showed similar results, the indicators of validity were higher in SAPA than AgNOR count. CONCLUSION: Although, AgNOR count and SAPA score gave similar results, but the indicators of validity were higher in SAPA score than AgNOR count.     

AgNOR定量分析在细胞学辅助诊断中的意义

目的利用核仁组成区技术(AgNOR)区分良性、中至重度不典型增生及恶性细胞,并探讨AgNOR分型与癌细胞类型的关系.方法应用胶银染色技术检测439例细胞学涂片.结果良性细胞均数(2.96±0.76)与恶性细胞均数(9.90±2.43)差异有极显著性(P＜0.001),说明产生假阳性机会很小.良性细胞与中度不典型增生细胞均数(3.90±0.66)和重度不典型增生细胞均数(5.88±0.98)差异有极显著性(P＜0.001),有助于识别可疑癌细胞.AgNOR在腺癌细胞中颗粒粗大、色深;在鳞癌及小细胞癌中颗粒多为弥散型,细小、色浅.AgNOR分型与癌细胞分型有显著相关性(P＜0.001).用AgNOR计算标准判断良恶性其敏感性为96.4%,特异性为95.5%.结论细胞学普通染色诊断困难时,AgNOR银染技术可以辅助诊断.     

Nucleolar organizer regions in Spitz nevi and malignant melanomas

Nucleolar organizer regions (NOR) are loops of DNA that transcribe ribosomal RNA; they can be easily identified in formalin fixed paraffin embedded tissue using a silver (Ag) method. It has been suggested that the number of AgNOR per cell can differentiate between benign and malignant melanocytic lesions of skin. We have studied 29 Spitz nevi (SN) and 39 invasive malignant melanomas (MM) by the same silver method. SN showed between 1.0 and 1.6 AgNOR per cell with a mean of 1.2. MM counts ranged from 1.2 to 4.2 with a mean of 2.0. It is concluded that the AgNOR method cannot reliably differentiate SN from MM; however, a count of more than 2.0 AgNOR per cell would favor a diagnosis of MM rather than SN.     

AgNOR计数对宫颈良恶性病变的鉴别诊断意义

目的 :探讨核仁形成区嗜银蛋白 (AgNOR)计数对宫颈良恶性病变鉴别诊断价值。方法 :采用AgNOR银染法对比 15例宫颈鳞癌与 2 2例慢性宫颈炎的AgNOR计数。结果 :宫颈鳞癌与宫颈良性病变的AgNOR计数差异有显著性 (P <0 0 1) ,宫颈不典型增生AgNOR计数也较宫颈良性增生上皮增高(P <0 0 5)。结论 :宫颈良恶性病变的AgNOR计数明显不同 ,此方法简便易行 ,可用于宫颈良恶性病变的鉴别。     

乳腺肿瘤细胞TrfR表达与AgNOR计数的相互关系及临床应用价值

收集57例乳腺肿块穿刺或印片标本,用PAP法进行免疫化学染色观察TrfR阳性细胞和核仁组成区嗜银蛋白染色计算AgNORs颗粒数。结果提示:乳腺癌与非癌组织之间OKT_9,及AgNORs数的差别非常显著(P<0.01),而乳腺纤维腺瘤与腺病其差别无意义(P>0.05),OKT_9阳性与阴性乳腺癌之间AgNORs数的差别是显著的(P<0.05),呈正比关系。我们认为AgNORs和TrfR检测,对判断乳腺细胞其良恶性有十分重要意义,能作为细胞穿刺的辅助诊断手段。     

Increased risk of cancer in the descendants of syrian hamsters exposed prenatally to diethylnitrosamine (DEN)

Transmission of site-specific tumorigenicity (papillomas in larynx and trachea) of diethylnitrosamine (DEN) to the 2 subsequent generations (F₁ and F₂) was studied using an out-bred strain (Han:AURA) of pregnant Syrian golden hamsters (P generation), which were treated i.p. with 10 mg/kg b.w. of DEN on day 12, 13 or 14 of gestation. Laryngotracheal papillomas were induced by DEN in the P and F₁ generations only, while these tumours did not occur in the F₂ generation. Spontaneously occurring tumours, including uterine adenocarcinomas, lymphomas, and laryngotracheal neuro-endocrine cell tumours, were observed at higher incidences among the F₂ animals derived from the P generation hamsters treated with DEN only on day 13 or 14 of gestation. In the same animals, the ratio of malignant to benign tumours was considerably higher than in controls. In addition, the F₂ hamsters derived from the DEN-treated P generation showed more frequent multiple organ involvement in tumorigenesis than the F₂ controls. Several uncommon malignant tumours were detected in the F₂ offspring, possibly the result of damage caused to germ cells by the prenatal exposure of F₁ Syrian hamsters to DEN.     

Prognostic value of histobiological factors (malignancy grading and AgNOR content) assessed at the invasive tumour front of oral squamous cell carcinomas.

Tumour cells at the invasive front of carcinomas have been found to differ substantially from the rest of tumour cells in a variety of human cancers. The present multivariate survival analysis of 94 oral squamous cell carcinomas (OSCCs) revealed that both the argyrophilic nucleolar organizer regions-associated protein (AgNOR) content of invading tumour cells and a multiparametric histopathological tumour front grade were significantly and independently associated with tumour-related death, irrespective of conventional Broders'' grade and clinical stage of the tumours. High tumour front scores and AgNOR content at the invasive OSCC front thus seem to reflect increased malignant potential. Proliferative activity, assessed by standardized AgNOR analysis, most probably represents one of the biological features underlying the usefulness of evaluating the invasive tumour front.     

Nucleolar organiser regions as indicators of post-surgical prognosis in canine spontaneous mast cell tumours

The average number of nucleolar organiser regions per cell has previously been shown to correlate well with histological grading techniques for a variety of neoplasms in man, and may thus be of value as an aid to post-surgical prognosis. In this study 50 spontaneously arising, subcutaneous canine mast cell tumours were graded and the histological grade compared with the mean AgNOR count. For well differentiated neoplasms the mean count was 1.4 per cell compared with 6.3 for poorly differentiated neoplasms, while tumours of intermediate differentiation had a mean count of 3.2 per cell. Subsequent follow up studies revealed that the AgNOR count was an accurate prognostic indicator, 73% of dogs with a high mean count (greater than 4.9) being destroyed from tumour related disease compared with 33% with an intermediate count (1.7-4.8). No dog with a count of less than 1.7 has been destroyed because of tumour recurrence to date and the AgNOR count has proved to be a better and more objective prognostic indicator than either histological tumour grade or mitotic index. Since most dogs which develop recurrent mast cell tumours do so within 6 months of initial surgery, an assessment of the predictive value of AgNORs can be obtained more quickly in canine tumours than for comparable human neoplasms.     

Expression profile of skin papillomas with high cancer risk displays a unique genetic signature that clusters with squamous cell carcinomas and predicts risk for malignant conversion

Chemical induction of squamous tumors in the mouse skin induces multiple benign papillomas: high-frequency terminally benign low-risk papillomas and low-frequency high-risk papillomas, the putative precursor lesions to squamous cell carcinoma (SCC). We have compared the gene expression profile of twenty different early low- and high-risk papillomas with normal skin and SCC. Unsupervised clustering of 514 differentially expressed genes (P<0.001) showed that 9/10 high-risk papillomas clustered with SCC, while 1/10 clustered with low-risk papillomas, and this correlated with keratin markers of tumor progression. Prediction analysis for microarrays (PAM) identified 87 genes that distinguished the two papilloma classes, and a majority of these had a similar expression pattern in both high-risk papillomas and SCC. Additional classifier algorithms generated a gene list that correctly classified unknown benign tumors as low- or high-risk concordant with promotion protocol and keratin profiling. Reduced expression of immune function genes characterized the high-risk papillomas and SCC. Immunohistochemistry confirmed reduced T-cell number in high-risk papillomas, suggesting that reduced adaptive immunity defines papillomas that progress to SCC. These results demonstrate that murine premalignant lesions can be segregated into subgroups by gene expression patterns that correlate with risk for malignant conversion, and suggest a paradigm for generating diagnostic biomarkers for human premalignant lesions with unknown individual risk for malignant conversion.     

胃良,恶性上皮核仁组成区嗜银蛋白定量研究

使用胶银染色技术对88例胃良、恶性病变及正常胃粘膜进行核仁组成区定量研究。结果表明:慢性胃溃疡之修复性增生上皮及癌旁粘膜平均每核含AgNOR量及AgNOR异形率均高于正常胃粘膜上皮(P<0.05),慢性胃溃疡增生上皮及癌旁粘膜上皮两者之间无明显差异。高分化腺癌与正常胃粘膜上皮、慢性胃溃疡修复上皮及癌旁粘膜上皮相比,平均每核含AgNOR量、AgNOR异形率及AgNOR颗粒在核内的大小及位置分布等均差异非常显著(P<0.01),提示AgNOR定量研究对区分胃良、恶性上皮有重要参考价值。     

DNA与AgNOR在甲状腺良、恶性病变中的定量研究

本研究采用Ｆｅｕｌｇｅｎ染色和ＡｇＮＯＲ银染色，检测ＤＮＡ及ＡｇＮＯＲ物质，发现：ＤＮＡ含量Ⅲ级为甲状腺癌组＞非典型结节组＞结甲肿组＞对照组，而Ⅰ级则反之；ＡｇＮＯＲ均数为甲状腺癌组（９．２７±２：７２）＞非典型结节组（４．７８±１．６６）＞结甲肿组（３．１６±１．３８）＞对照组（３．０８±０．４１）；甲状腺癌组与各组间差异有显著意义（Ｐ＜０．０１），非典型结节组与结甲肿组、对照组间差异也有显著意义（Ｐ＜０．０１），而结甲肿组与对照组间则无差异（Ｐ＞０．０５）。作者认为Ⅲ级ＤＡＮ含量的细胞出现频率越高或／和ＡｇＮＯＲ均数越高，恶性的可能性就越大或分化程度越低；本研究提示ＡｇＮＯＲ的均数与ＤＮＡ含量具有显著的相关性，并与细胞增生的程度呈正比，故对鉴别良、恶性肿瘤及交界性病变有肯定价值。     

胃癌和胃粘膜异型增生核仁组织区嗜银蛋白定量和形态研究

目的探讨核仁组织区嗜银蛋白（ＡｇＮＯＲ）染色技术在胃癌、癌前病变、慢性胃炎中鉴别诊断价值和意义。方法应用ＡｇＮＯＲ染色技术，观察１３２例胃良恶性病变和癌前病变细胞核中ＡｇＮＯＲ颗粒含量和形态。结果胃癌、胃粘膜异型增生、慢性胃炎三组间细胞核内ＡｇＮＯＲ颗粒均数差异非常显著（Ｐ＜０．０１），正常胃壁粘膜与慢性胃炎ＡｇＮＯＲ颗粒均数无明显差异（Ｐ＞０．０５）。胃癌和胃粘膜异型增生ＡｇＮＯＲ颗粒形态以弥散型为主，而慢性胃炎和正常胃粘膜以核仁型为主。结论细胞核内ＡｇＮＯＲ颗粒含量和分型对于区别胃良恶性及癌前病变有重要参考价值。     

Sequential study of gamma-glutamyltransferase during complete and two-stage mouse skin carcinogenesis

The histochemical pattern of gamma-glutamyltransferase (GGT) was studied in benign and malignant tumors produced by two different experimental protocols, two-stage carcinogenesis and complete carcinogenesis. Six percent of all papillomas produced by two-stage carcinogenesis were GGT positive, whereas 14% of benign tumors produced by complete carcinogenesis exhibited GGT-positive areas. The incidence of GGT-positive papillomas in the two-step carcinogenesis protocol increased up to wk 28 of treatment. After 32 wk, the incidence decreased abruptly, coinciding with an abrupt increase in the incidence of squamous cell carcinomas. On the other hand, the incidence of GGT-positive benign tumors produced during the course of complete carcinogenesis increased gradually up to wk 32 of treatment, coinciding with the increased incidence of squamous cell carcinomas. The incidence of GGT-positive keratoacanthomas and GGT-positive papillomas produced with the complete carcinogenesis protocol exhibited different patterns, suggesting different histogenesis and biological behavior of these two types of tumors. In addition, the labeling index of GGT-positive areas was lower (17 +/- 3%) than that of the GGT-negative areas (41 +/- 0.18%) of the same papillomas, indicating that the presence of GGT may be related to abnormal keratocyte differentiation rather than to proliferative changes.     

Investigations on the carcinogenic burden by air pollution in man. Intratracheal instillation studies with benzo[a]pyrene in a mixture of Tris buffer and saline in Syrian golden hamsters.

Syrian golden hamster received intratracheal instillations of 0.125, 0.25, 0.50 or 1.0 mg benzo[a]pyrene (B[A]P) in a mixture of Tris buffer and physiological saline once weekly for life. Papillary polyps, squamous cell papillomas and carcinomas developed in both the larynx and trachea. In addition, bronchiogenic adenomas, adenocarcinomas and squamous cell carcinomas were induced in the lung. The highest incidence of respiratory tract tumours (83%) was seen in hamsters receiving 0.25 mg B[a]P. The results of these investigations are statistically evaluated and discussed.     

P16INK4A immunostaining is a strong indicator for high‐risk‐HPV‐associated oropharyngeal carcinomas and dysplasias,but is unreliable to predict low‐risk‐HPV‐infection in head and neck papillomas and laryngeal dysplasias

Human papillomavirus (HPV) is a risk factor for the development of benign and malignant mucosal head and neck lesions. P16^INK4A is often used as a surrogate marker for HPV‐infection, although there is still controversy with respect its reliability. Our aim was to determine if p16^INK4A overexpression can accurately predict both high‐risk and low‐risk‐HPV‐presence in (pre)malignant and benign head and neck lesions. P16^INK4A immunohistochemistry was performed on paraffin‐embedded tissue sections of 162 oropharyngeal squamous cell carcinomas (OPSCC), 14 tonsillar and 23 laryngeal dysplasias, and 20 tonsillar and 27 laryngeal papillomas. PCR, enzyme‐immunoassay and FISH analysis were used to assess HPV‐presence and type. Of the 162 OPSCC and 14 tonsillar dysplasias, 51 (31%) and 10 (71%) were HPV16‐positive, respectively. All tonsillar papillomas were HPV‐negative and four laryngeal dysplasias and 26 laryngeal papillomas were positive for HPV6 or ?11. P16^INK4A immunohistochemistry revealed a strong nuclear and cytoplasmic staining in 50 out of 51 HPV16‐positive and 5 out of 111 HPV‐negative OPSCC (p < 0.0001) and in all HPV16‐positive tonsillar dysplasias, whereas highly variable staining patterns were detected in the papillomas and laryngeal dysplasias, irrespective of the HPV‐status. In addition, the latter lesions generally showed a higher nuclear than cytoplasmic p16^INK4A immunostaining intensity. In conclusion, our data show that strong nuclear and cytoplasmic p16^INK4A overexpression is a reliable surrogate indicator for HPV16 in OPSCC and (adjacent) dysplasias. For HPV6 or ?11‐positive and HPV‐negative benign and premalignant lesions of the tonsil and larynx, however, p16^INK4A immunostaining is highly variable and cannot be recommended to predict HPV‐presence.     

Bax expression has prognostic significance that is enhanced when combined with AgNOR counts in glottic carcinomas.

Using nucleolar organizer regions (NORs) as a proliferative marker and Bax expression as a marker for apoptosis, we have studied the individual and combined prognostic significance of these markers. Successive sections of diagnostic, formalin-fixed and paraffin-embedded specimens from 69 patients with T1-4 tumours were stained with a rabbit anti-human Bax polyclonal antibody and silver nitrate for visualization of NORs (AgNORs). After classification for staining intensity and the percentage of Bax expression, a final score resulting in four classes of increasing Bax expression was obtained. AgNOR counts were expressed as mean counts (mAgNOR) and the percentage of tumour nuclei with more than one AgNOR (pAgNOR>1). Both AgNOR parameters were grouped in three classes with increasing values. Low Bax scores correlated significantly with poor prognosis (P = 0.0106). For mAgNOR and pAgNOR>1, high values correlated with poor prognosis (P = 0.0185 and P = 0.0003 respectively). A combined parameter, for which the Bax score was subtracted from the AgNOR scores, appeared to be statistically stronger than the individual parameters (P < 0.0001). Both Bax expression and AgNOR scores, and in particular the combination of these parameters, appear to be strong prognostic markers in glottic squamous cell carcinomas.     

滋养细胞肿瘤核仁蛋白组成区研究

 运用核仁蛋白组成区嗜银蛋白染色术检测了5例正常胎盘,1例合体细胞子宫内膜炎,22例葡萄胎,10例恶性葡萄胎,2例胎盘部位中间型滋养细胞肿瘤和19例绒癌,发现恶性滋养细胞肿瘤的AgNOR/核均数明显高于良性滋养细胞肿瘤(P<0.001),细胞滋养细胞中的AgNOR/核均数亦明显高于合体滋养细胞(P<0.001).同时发现良恶性滋养细胞肿瘤的AgNOR颗粒分布类型也不同,良性病变以弥漫型为主,而恶性病变以颗粒型为主,表明主要用于细胞遗传学的AgNOR技术在肿瘤病理学上有广泛的应用价值.     

The high-risk benign tumor: evidence from the two-stage skin cancer model and relevance for human cancer

Benign tumors that form following chemical initiation and promotion in the mouse skin can be grouped into two classes. The majority of papillomas do not progress to squamous cell carcinoma (SCC), and these are designated as low-risk or terminally benign papillomas. In contrast, a much smaller group forms the true precursor to the SCC, and these have a significantly higher frequency and rate of malignant conversion than the bulk of low-risk papillomas. In standard two-stage carcinogenesis studies both tumor types are present, but grossly indistinguishable. Here we describe properties and potential origins of high-risk papillomas and discuss the relevance of this model for certain human cancers with defined premalignant states.