胃癌及癌前病变细胞核DNA含量和核仁组成区嗜银蛋白计数研究

本文通过胃癌及癌前病变细胞核DNA含量测定,AgNOR(核仁组成区嗜银蛋白染色)计数的研究,旨在探讨其对胃癌及癌前病变分级诊断中的意义。     

大肠癌和大肠腺瘤DNA含量与细胞增殖活性检测的临床意义

分子生物学及细胞生物学的研究表明,恶性肿瘤最重要的生物学特征为肿瘤细胞异常增殖和分化不良、浸润性生长和远处转移以及细胞恶性特征的遗传性.以上3点癌细胞共同生物学特征的出现,均以细胞核DNA含量和(或)染色体倍体与功能等微细异常为物质基础.因此,测量和分析癌前病变及癌组织中细胞核DNA含量与倍体对恶性肿瘤的病理诊断、恶性程度判定、疗效评估、预测预后均具有重要价值.本研究应用流式细胞术(FCM)检测了经手术或肠镜取得的对照组(正常大肠黏膜)、大肠腺瘤(绒毛状腺瘤、管状腺瘤)和大肠癌组织,检测了它们的DNA倍体、DNA含量和细胞的增殖活性,并进行了临床分析.     

细胞核DNA含量测定和AgNOR颗粒计数对预测卵巢囊腺癌患者预后的意义

背景与目的:以往认为,细胞核DNA含量和核仁组成区嗜银蛋白(silver-stainingnucleolarorganizerregion,AgNOR)计数在某些肿瘤中有重要的预后意义,但对卵巢囊腺癌患者的预后意义尚不清楚。为此,本研究探讨该两项细胞增殖参数对预测卵巢囊腺癌患者预后的价值。方法:对42例卵巢囊腺癌切除术的组织标本采用图像分析仪测定核DNA含量和AgNOR颗粒计数,将结果与预后情况进行分析。结果:单因素生存率分析显示,差异有统计学意义的临床变量为FIGO分级、分化程度、残余肿瘤大小、肿瘤转移灶、淋巴结转移情况和年龄、异倍体、DNA指数、AgNOR颗粒计数(P<0.05)。多因素生存率分析显示,DNA指数>1.4、FIGO分级、分化程度和残余肿瘤大小是卵巢囊腺癌的具有统计学意义的预后预测因子。结论:对于卵巢囊腺癌患者来说,细胞核DNA指数可能是有用的预后独立预测因子。     

甲状腺滤泡性肿瘤DNA含量与AgNOR计数研究

目的：探讨DNA含量与AgNOR对甲状腺良恶性滤泡性肿瘤的鉴别诊断价值。方法：应用流式细胞术和胶体银染技术对9例正常甲状腺组织和36例滤泡性肿瘤（22例腺瘤和14例腺癌）进行DNA含量分析和AgNOR计数。结果：14例滤泡性癌中，11例为DNA异倍体，而22例滤泡性腺癌仅1例为DNA异倍体，且伴有不典型增生。AgNOR由正常甲状腺组织、滤泡性腺癌至滤泡性癌逐渐增加，相互之间有显著性差异（P＜0．05）；但各组间AgNOR计数有重叠，且DNA含量与AgNOR无明显相关性。结论：尽管AgNOR染色方法简便。经济，但在区别良恶性滤泡性肿瘤方面，DNA含量异常更具意义。     

辐射损伤骨髓造血细胞的DNA含量及AgNOR定量分析

LACA小鼠经60 Coγ射线全身一次照射 ,对骨髓组织切片和骨髓细胞悬液涂片 ,分别采用改良的Feulgen染色及核仁组成区相关蛋白 (AgNOR)染色 ,利用图像分析技术对在不同照射剂量、不同时间点的造血细胞核内DNA及AgNOR含量进行了定量分析。结果显示各照射组在2 5 - 7 0Gy剂量、 6h～ 4周范围内 ,骨髓均出现明显损伤及损伤后重建现象 ,且剂量越大 ,损伤越重 ,恢复亦较慢 ;其中以 5 5Gy组骨髓细胞核内DNA和AgNOR含量变化最为典型 ,在损伤早期进行性减少 ,而在恢复期出现持续性增加 ,直至恢复正常。结果提示 :骨髓造血细胞核内AgNOR及DNA含量的变化可以作为反映骨髓辐射损伤与修复程度的定量指标。     

VEGF在甲状腺癌及转移淋巴结中的表达及与DNA倍体、AgNOR的关系

目的研究血管内皮生长因子（VEGF）在甲状腺癌及其转移淋巴结中的表达及与癌细胞DNA倍体、核仁形成区（AgNOR）的关系。方法应用免疫组织化学SABC法检测136例甲状腺癌及淋巴结中VEGF的表达，同时进行细胞DNA含量及AgNOR自动图像分析检测，10例正常甲状腺组织作对照。结果10例正常甲状腺组织VEGF（一），136例甲状腺癌组织中VEGF阳性表达72．79％（99／136），淋巴结中VEGF阳性表达97％（132／136）。DNA倍体平均值及AgNOR计数5年以上生存组病例明显低于5年以内死亡组（P＜O．01，P＜O．05）。结论检测VEGF、DNA倍体及Ag－NOR有助于评估甲状腺癌的生物学行为及预后。     

喉乳头状瘤及癌的细胞核形态计量分析和DNA含量相关性研究

本文采用图像分析系统对喉乳头状瘤伴上皮不典型增生及喉鳞状细胞癌进行了细胞核形态定量学研究 :同时检测了细胞核的DNA含量。结果显示 :随鳞状上皮不典型增生程度加重及喉癌的出现 ,细胞核形态学参数 (核面积、核长径、核周长及核圆度 )逐渐增加 ,重度不典型增生和癌变之间存在明显界限 ;并且核面积与DNA含量之间具有明显的正相关关系。提示 :本方法对早期发现喉的癌前病变及癌提供了一种客观有效的形态学诊断依据 ,具有一定的临床实用价值。     

脑膜瘤MR征象与DNA含量的相关性研究

目的:研究脑膜瘤磁共振(magnetic resonance,MR)征象与肿瘤细胞核DNA含量的相关性,从MRI影像学角度评价脑膜瘤的生物学行为及细胞增殖潜能.方法:45例脑膜癌患者,应用细胞图像分析仪,采用吸光度法对脑膜瘤细胞核DNA含量和倍性水平进行检测,所得数据资料与MR征象进行相关性分析.结果:脑膜瘤瘤周水肿、肿瘤外形不规则、瘤脑边界模糊毛糙3种MR征象的DNA含量及倍性水平与无瘤周水肿、肿瘤外形光滑和瘤周边界清晰的脑膜瘤比较,差异均有统计学意义(P均<0.01),这3种征象的MRI得分与DNA含量相关(r=0.696,P=0.000).结论:脑膜瘤MR图像上,有瘤周水肿、不规则肿瘤外形、瘤脑边界模糊毛糙者,其DNA含量及异倍体率较高,可作为预测脑膜瘤增殖潜能和恶性生物学行为的影像学指标.     

乳腺癌DNA倍体与ER表达的研究

目的 探讨乳腺癌DNA倍体与ER的相关性和生物学行为。方法 本文应用流式细胞术(FCM)检测了30例乳腺癌的细胞核DNA含量,同时用免疫组化PAP法测定了肿瘤细胞中雌激素受体(ER)的表达,以探讨其二者的相关性和生物学行为。结果 乳腺癌DNA含量63.3％(19/30)为异倍体,37.7％(11/30)为二倍体或近二倍体。DNA含量与ER呈负相关(P<0.01)。异倍体在ER阳性者达78.57％(11/14)。结论 DNA含量与临床分期相关性显著,即与肿瘤大小,分化程度及预后相关密切。也提示ER表达DNA含量有一定相关性。DNA含量是反应肿瘤生物学行为较正确、客观的生物学指标。     

AgNOR对结肠粘膜病变良恶性鉴别及意义

目的应用核仁形成区相关嗜银蛋白(AgNOR)对不同结肠粘膜病变的细胞核内AgNOR的数量、形态、大小和分布规律进行观察和对比分析.方法切取结肠粘膜组织,结肠乳头状腺瘤,结肠癌标本用10%中性福尔马林固定,酒精脱水等处理.然后按AgNOR药盒使用说明进行染色.结果结肠粘膜细胞核内AgNOR均数在1.84±0.69,结肠乳头状腺瘤3.58±1.舵,结肠癌6.53±1.94,三者之间显著差异.结论AgNOR的观察与研究反映了结肠粘膜各病变时,细胞核增生程度对结肠良恶性肿瘤的鉴别,尤其是边界性肿瘤的鉴别是具有研究价值的.     

CORRELATIONSHIP BETWEEN CELLULAR DNA AND AgNOR PROTEIN CONTENT IN THE DEVELOPING COURSE OF COLORECTAL ADENOCARCINOMA

谢尧，屈汉廷ＣＯＲＲＥＬＡＴＩＯＮＳＨＩＰＢＥＴＷＥＥＮＣＥＬＬＵＬＡＲＤＮＡＡＮＤＡｇＮＯＲＰＲＯＴＥＩＮＣＯＮＴＥＮＴＩＮＴＨＥＤＥＶＥＬＯＰＩＮＧＣＯＵＲＳＥＯＦＣＯＬＯＲＥＣＴＡＬＡＤＥＮＯＣＡＲＣＩＮＯＭＡ￥ＸｉｅＹａｏ；ＱｕＨａｎｔｉｎｇ...     

胃癌前病变AgNOR面积及形态与其DNA含量的对照研究

本文应用计算机图象分析系统对１０例肠化、３０例不同程度异型增生的胃粘膜，１０例正常胃粘膜和１０例高分化腺癌细胞核内ＡｇＮＯＲ蛋白的面积及其颗粒的形状进行了定量，并测定了其ＤＮＡ含量，结果显示，随胃粘膜病变程度的加重，其核内ＡｇＮＯＲ蛋白与ＤＮＡ含量依次增加，而ＡｇＮＯＲ颗粒的形状因子呈递减趋势。ＡｇＮＯＲ蛋白面积／核和其颗粒的形状因子与ＤＮＡ含量均有良好的线性相关关系，前者为正相关（ｒ＝０．９９Ｐ＜０．００１），后者为负相关（ｒ＝－０．９５２Ｐ＜０．００１）。我们认为，ＡｇＮＯＲ面积及其颗粒的形状可以反映出胃粘膜的病变程度。可作为胃癌前病变诊断及病情监测的有用指标。     

Cytophotometric DNA content and argyrophilic nucleolar organiser regions of oesophageal carcinoma.

The cytophotometric DNA content and the argyrophilic nucleolar organiser regions (AgNORs) of biopsy specimens taken before undergoing any treatment were examined in 91 surgically treated oesophageal carcinoma cases. There was a significant linear dependence between the mean DNA content and the number of AgNOR per nucleus (AgNOR number) (r = 0.615, P < 0.001). The DNA distribution pattern and the range of the AgNOR number also showed a significant correlation (P < 0.01). Twenty three of 28 cases with a low AgNOR number (< 4) were then determined to have a diploid pattern (type II), while 17 out of 22 cases with a high AgNOR number (> or = 6) had high ploidy values (type IV). The patients with a type II DNA distribution pattern and a low AgNOR number thus showed a good post-operative course with a 5 year survival rate of 55.2%, whereas no patients survived over 4 years among the 17 cases with both a type IV DNA pattern and a high AgNOR number (P < 0.001). These data thus demonstrate the close relationship between cytophotometric DNA content and AgNOR number and suggest that the combined detection of these two parameters, using biopsy specimens, should be of benefit in making an accurate preoperative evaluation of prognosis for patients with oesophageal carcinoma.     

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。     

用形态测量方法研究卵巢上皮性肿瘤的诊断及预后

背景与目的:形态测量学在肿瘤的病理诊断及预后评估中日渐重要。本文探讨图像分析法在卵巢上皮性肿瘤的诊断及预后中的意义。方法:联合应用图像分析、AgNOR计量分析及DNA含量测定3种形态测量学方法,对110例原发性卵巢上皮性肿瘤的诊断及预后进行研究,选用的参数为核面积(nucleararea,NA)、核周长(nuclearperimeter,NP)和核形状因子(nuclearformfactor,NFF)、DNA含量(DNAcontent,DC)、DNA指数(DNAindex,DI)、G0/G1期细胞百分率(percentageofG0/G1phases,P2c)、高倍体细胞百分率(percentageofDNAmultiploid,P>4c),并于光镜下进行AgNOR计数。结果:(1)良性、交界性及恶性3组卵巢上皮性肿瘤相比较,NA、NP、NFF、DC、DI及AgNOR计数在各组间差异有极显著性意义(P<0.01或P<0.001)。(2)形态测量学结果与预后关系研究表明,卵巢上皮性癌患者生存≥5年及<5年两组病例中,NA、NP、NFF参数两组间差异有显著性或极显著性意义(P<0.05或P<0.01),不同的图像分析结果提示其预后不同。AgNOR计数与上皮性卵巢癌患者的生存时间呈负相关(r=-0.73,P<0.001)。DNA高倍体含量与上皮性卵巢癌患者生存时间亦呈负相关(r=-0.75,P<0.001)。结论:形态测量学可为不同性质的卵巢上皮性肿瘤的诊断和预后提供可靠的客观依据。     

Gains and amplifications of c-myc, EGFR, and 20.q13 loci in the no dysplasia-dysplasia-adenocarcinoma sequence of Barrett's esophagus

The progression of Barrett's esophagus to esophageal adenocarcinoma is often characterized by the accumulation of genetic abnormalities. The goal was to evaluate the copy number alterations of several oncogene loci, including 7p12 [epidermal growth factor receptor (EGFR)], 8q24 (c-myc), and 20q13 in the sequence of no dysplasia-dysplasia-adenocarcinoma of Barrett's esophagus. Fluorescence in situ hybridization with DNA probes for the centromeric region of chromosome 7 and the locus-specific regions of 7p12 (EGFR), 8q24 (c-myc), and 20q13 was applied on 99 brush cytology specimens of patients with Barrett's esophagus with different stages of dysplasia or esophageal adenocarcinoma. Gains (3-4 copies) of chromosome 17, 8q24 (c-myc), and 20q.13 loci were found in the low frequencies in nondysplastic Barrett's esophagus. Their frequencies increased with the stage of dysplasia and reached a high incidence in esophageal adenocarcinoma. Amplification (>4 copies) of at least 1 of the loci was observed in 14% of high-grade dysplasia and increased to 50% in esophageal adenocarcinoma (P = 0.015). The most frequently amplified locus was c-myc (18%), followed by 20q13 (13%) and EGFR (11%) in the high-grade dysplasia/esophageal adenocarcinoma cases. High amplification levels (>10 copies) of the loci were more frequent in esophageal adenocarcinoma (72%) compared with high-grade dysplasia (20%; P = 0.049). Amplifications of the c-myc, EGFR, and 20q12 loci may serve as diagnostic markers to identify patients with Barrett's esophagus with high-grade dysplasia or esophageal adenocarcinoma. Gains of the loci might be of value as prognostic markers because they are already present in nondysplasia cases and may precede the later event of the amplification as observed in high-grade dysplasia and esophageal adenocarcinoma.     

Oxidative stress has a role in malignant transformation in Barrett's oesophagus

Mechanisms underlying the development of oesophageal adenocarcinoma are poorly understood. To discover the role of oxidative stress and radical scavenger capacity in the malignant transformation of Barrett's oesophagus, we measured myeloperoxidase activity, superoxide dismutase activity, glutathione content and total aromatic DNA adducts. Mucosal specimens came from 52 patients in 6 groups: symptomatic gastro-oesophageal reflux disease (GORD) without and with endoscopic oesophagitis, Barrett's epithelium without and with dysplasia, adenocarcinoma in the oesophagus and controls. In the GORD-oesophagitis-metaplasia-dysplasia-adenocarcinoma sequence, glutathione content was progressively lower and myeloperoxidase activity higher than in controls, plateauing at Barrett's epithelium without dysplasia. Only in Barrett's epithelium with dysplasia was SOD activity significantly increased. In all patient groups, DNA adduct levels were significantly higher than the control level. Though these levels between patient groups did not differ significantly, the level was highest in Barrett's epithelium without dysplasia and progressively lower in Barrett's with dysplasia and adenocarcinoma. Pooled data showed a negative correlation between glutathione content and DNA adducts (-0.28, p = 0.05). Simultaneous formation of DNA adducts, increased myeloperoxidase-related oxidative stress, decreased antioxidant capacity (glutathione content) and the negative correlation between glutathione content and DNA adducts in the GORD-oesophagitis-metaplasia-dysplasia-adenocarcinoma sequence of Barrett's oesophagus indicate a role in the pathogenesis and malignant transformation related to oxidative stress.     

Nuclear DNA content and nucleolar organizer regions in colorectal cancer

The prognostic value of nuclear DNA content and argyrophilic nucleolar organizer regions (AgNOR) is still controversial in colorectal cancer. Sixty patients with colorectal cancer were studied by flow cytometric DNA analysis and AgNOR measurement, and their prognostic significance was tested. DNA index was closely linked to depth of invasion and lymph node metastasis, while AgNOR count did not correlate with such parameters. The survival curve was stongly influenced by depth of invasion, lymph node metastasis, and Dukes' stage but was not affected by DNA ploidy and AgNOR count. These results indicate that neither DNA ploidy nor AgNOR count correlates with survival of patients, although DNA ploidy is linked to progression of colorectal cancer.     

9p21 Deletion in the diagnosis of malignant mesothelioma in serous effusions additional to immunocytochemistry,DNA‐ICM,and AgNOR analysis

BACKGROUND

The diagnosis of malignant mesothelioma (MM) in serous effusions is difficult but may be achieved by the application of adjuvant methods.

METHODS

The authors cytologically diagnosed 33 effusions as suspicious or positive for MM cells by using DNA‐image cytometry (DNA‐ICM), immunocytochemistry and AgNOR analysis. The authors further detected 9p21 deletions by chromosomal fluorescence in situ hybridization (FISH). In addition, 31 cases of metastatic carcinomas and 39 of tumor cell‐negative effusions were investigated. All diagnoses were confirmed by histologic and/or clinical follow‐up.

RESULTS

DNA aneuploidy was found in 71% of MMs, 100% of metastatic carcinomas, and in none of the negative effusions. Calretinin was positive in 100% of MMs, in none of the metastatic carcinomas, and in 94.9% of negative effusions. BerEP4 showed positivity in 15.6% of MMs, 87.1% of metastatic carcinomas, and in none of the negative effusions. With AgNOR analysis, 89.3% of MMs and 96.7% of metastatic carcinomas showed ≥2.5 AgNOR dots as satellites and ≥4.5 as total AgNOR counts. 9p21 deletions were demonstrated in 90.9% of MM cases, 45.2% of metastatic carcinomas, and in none of the negative effusions. By cytology alone, 81.8% of MMs were identified unequivocally. Addition of DNA‐ICM improved the prevalence of tumor cell detection to 87.9% and of AgNOR analysis to 97%. The introduction of 9p21 deletions by FISH improved this prevalence to 100%.

CONCLUSIONS

Because of these results, the authors propose the sequential application of immunocytochemistry, DNA‐ICM, and AgNOR analysis to establish a cytological diagnosis of malignant mesothelioma in serous effusions. In persistent doubtful diagnoses, the authors recommend fluorescence in situ hybridization to analyze the 9p21 deletion. Cancer (Cancer Cytopathol) 2008. © 2008 American Cancer Society.