67例结肠癌DNA倍体和Ag—NOR计数分析对判断患者预后价值的初步探讨

对６７例结肠癌石蜡包埋组织材料做ＤＮＡ倍体水平分析和Ａｇ—ＮＯＲ计数。结果见：ＤＮＡ二倍体肿瘤５年、７年生存率分别为７８．０％（３２／４１）、３９．０％（１６／４１）；非二倍体肿瘤则为２６．９％（７／２６）、１１．５％（３／２６）（ｐ＜０．０１、ｐ＜０．０５）；Ａｇ─ＮＯＲ计数小于均值（６．９±１．０）组５年、７年生存率为８０．０％（２８／３５）、４０．０（１４／３５），大于均值组则为３４．４％（１１／３２）、１５．６％（５／３２）（ｐ＜０．０１、ｐ＜０．０５）。Ａｇ－ＮＯＲ计数小于均值组ＤＮＡ二倍体肿瘤占９７．１％（３４／３５）；大于均值组二倍体肿瘤占２１．９％（７／３２）（ｐ＜０．０１）。结果表明：ＤＮＡ倍体水平和Ａｇ－ＮＯＲ计数具有一致性，可以做为判断结肠癌预后指标。     

血卟啉衍生物对鼠肿瘤细胞DNA含量,AgNORs的影响

应用图像分析技术测定大鼠移植Ｗ２５６肿瘤单纯放射治疗组（对照组）和血卟啉衍生物并放射治疗组（实验组）肿瘤细胞的ＤＮＡ含量及ＡｇＮＯＲｓ计数，探讨血卟啉衍生物对恶性肿瘤放射治疗的增敏作用。结果显示实验组大鼠肿瘤细胞ＤＮＡ含量为（５．８４±１．２２）Ｃ，ＡｇＮＯＲｓ计数为２．４２±０．７３；对照组大鼠肿瘤细胞ＤＮＡ含量为（７．８３±０．６８）Ｃ，ＡｇＮＯＲｓ计数为５．３５±０．８４；前者ＤＮＡ含量及ＡｇＮＯＲｓ计数显著低于后者（ｔ＝６．３７，ｔ＝１１．７７，Ｐ＜０．０１）。表明血卟啉衍生物对恶性肿瘤放射治疗有明显的增敏作用     

宫颈良、恶性病变核仁组成区蛋白定量与宫颈癌预后的研究

应用核仁组成区蛋白嗜银（ＡｇＮＯＲ）技术，对１０９例宫颈良、恶性病变进行了ＡｇＮＯＲ定量研究。结果表明：１．慢性宜颈炎、宫颈非典型增生、宫颈鳞癌三种病变的ＡｓＮＯＲ均值有显著差异（Ｐ＜０．０１）。２．宫颈鳞癌组织学１级ＡｇＮＯＲ计数低于组织学Ⅱ级（Ｐ＜０．００１）。３．随访１０年以上的７３例宫颈鳞癌患者，核仁组成区蛋白高均值组（≥５）的患者生存时间显著短于低均值组（＜５）（Ｐ＜０．０５），１０年生存率分别为１８％与４６％。作者认为，ＡｇＮＯＲ计数可作为一种新的肿瘤定量指标，在宫颈良、恶性病变的鉴别诊断、选择最佳治疗方案、推测宫颈癌预后等方面具有重要的临床价值。     

紫外光血液辐照疗法对放射治疗鼠肿瘤细胞DNA含量、AgNORs计数的影响

应用多功能图像分析仪，检测了鼠Ｗ２５６移植性肿瘤细胞ＤＮＡ含量，并同时测定细胞ＡｇＮＯＲｓ计数，探讨紫外光血液辐照疗法（ＵＢＩ）对放射治疗的增敏作用。结果表明；光量＋放疗组肿瘤细胞ＤＮＡ含量为５．３４±１．０９Ｃ，ＡｇＮＯＲｓ计数为２．２４±０．４３；单纯放疗组肿瘤细胞ＤＮＡ含量为７．８３±０．８６Ｃ，ＡｇＮＯＲｓ计数为５．３５±０．８４；前者ＤＮＡ含量及ＡｇＮＯＲｓ计数显著小于后者（Ｐ＜０．０５）。表明ＵＢＩ对放疗恶性肿瘤有明显的增敏作用。     

甲状腺癌中VEGF表达与DNA含量、AgNOR关系的研究

目的研究血管内皮生长因子（ＶＥＧＦ）在甲状腺癌及其区域淋巴结中的表达及与癌细胞ＤＮＡ含量、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的ＶＥＧＦ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例正常甲状腺组织中未发现ＶＥＧＦ的表达,１３６例甲状腺癌组织中ＶＥＧＦ阳性表达７２．９７％（９９／１３６）,区域淋巴结中ＶＥＧＦ阳性表达９１％（１２４／１３６）。ＤＮＡ倍体平均值及ＡｇＮＯＲ５年以上生存组病例明显低于５年以内死亡组（Ｐ＜０．０１,Ｐ＜０．０５）。结论检测ＶＥＧＦ、ＤＮＡ含量及Ａｇ┐ＮＯＲ有助于评估甲状腺癌的生物学行为及预后。     

大肠癌细胞DNA、AgNORs定量研究──两者相关性及预后意义的探讨

应用ＡｇＮＯＲｓ染色技术，对６３例大肠癌的ＡｇＮＯＲｓ颗粒进行了计数研究。其中３８例同时进行肿瘤细胞核ＤＮＡ含量静态测定。结果表明，大肠癌细胞ＡｇＮＯＲｓ数量与其ＤＮＡ含量之间有明显相关性（Ｐ＜０．０ｌ）。ＤＮＡ含量和ＡｇＮＯＲｓ计数愈低，肿瘤分化愈好，患者预后愈好；反之，ＤＮＡ含量和Ａｇ－ＮＯＲｓ计数愈高、肝瘤分化愈差，患者预后愈差。表明ＡｇＮＯＲｓ计数，ＤＮＡ含量检测均可作为大肠癌分级、判断预后的参考指标。     

前列腺癌AgNOR计数与DNA含量测定的意义

作者应用银染色技术对３０例前列腺癌、２０例前列腺增生症组织中的核仁组成区相关嗜银蛋白（ＡｇＮＯＲ）进行检测，并采用图像分析技术测定其ＤＮＡ含量。结果显示，在前列腺癌组织中，ＡｇＮＯＲ计数与ＤＮＡ含量均明显高于前列腺增生症组织（Ｐ＜０．００１）；肿瘤分化越差，ＡｇＮＯＲ计数和ＤＮＡ含量越高，并且与患者预后相关（Ｐ＜０．００１）；ＡｇＮＯＲ计数与ＤＮＡ含量呈明显的正相关（Ｐ＜０．０１）。结果提示ＡｇＮＯＲ计数与ＤＮＡ含量可作为反映前列腺癌生物学行为的指标。     

PCNA和AgNOR在胃肠道平滑肌肿瘤中的表达研究

目的　探讨ＰＣＮＡ 和Ａｇ ＮＯＲ 在胃肠道平滑肌肿瘤组织中的表达及其与肿瘤良恶性的关系。方法　采用免疫组化ＳＰ 法和Ｈｏｗ ｅｌｌ 银染一步法检测６０ 例病理形态典型的胃肠道平滑肌肿瘤的ＰＣＮＡ 和Ａｇ ＮＯＲ 。结果　平滑肌肉瘤ＰＣＮＡ 指数明显高于平滑肌瘤,两者间有非常显著性差异（ Ｐ ＜ ０ ．０１） 。平滑肌肉瘤Ａｇ ＮＯＲ 平均含量（ 每个细胞内的平均颗粒数） 和颗粒直径（μｍ ） 分别为（８ ．２５ ±０ ．９２） μｍ 和（２ ．９０ ±０ ．２３） μｍ ,平滑肌瘤则分别为（２ ．８６ ±０ ．２９） μｍ 和（２ ．１２ ±０ ．１４） μｍ ,两者有非常显著性差异（ Ｐ＜ ０ ．０１） 。结论　ＰＣＮＡ 指数和Ａｇ ＮＯＲ 颗粒数及颗粒直径可作为鉴别胃肠道平滑肌肿瘤良恶性的客观指标之一。     

增殖细胞核抗原与AgNOR检测在前列腺癌临床中的意义

目的研究前列腺癌中增殖细胞核抗原（ＰＣＮＡ）表达及ＡｇＮＯＲ计数的意义。方法应用免疫组织化学方法和银染色技术检测前列腺癌和良性前列腺增生组织中ＰＣＮＡ表达和ＡｇＮＯＲ计数。结果ＰＣＮＡ增殖指数与ＡｇＮＯＲ计数在癌组织中均明显高于良性前列腺增生组织（Ｐ均＜０．００１）,且两者均与肿瘤组织学分级和预后有密切关系（Ｐ均＜０．０１和Ｐ均＜０．００１）;前列腺癌ＰＣＮＡ增殖指数与ＡｇＮＯＲ计数间存在非常显著的正相关（Ｐ＜０．０１）。结论ＰＣＮＡ增殖指数和ＡｇＮＯＲ计数结合分析,在鉴别前列腺良恶性病变和判断前列腺癌的恶性程度及预测患者预后方面具有十分重要的意义。     

喉癌及癌前病变核仁组成区银染和DNA含量测定及其相关研究

本文对５５例喉癌及癌前病变行Ａｇ－ＮＯＲｓ染色，并对其中部分病例用自动图象分析技术ＤＮＡ定量测定，发现Ａｇ－ＮＯＲＳ值喉癌＞不典型增生＞乳头状瘤，各组间差异有显著性（Ｐ＜０．０５）。ＤＮＡ含量测定ＳＣ细胞所占比例喉癌＞不典型增生＞乳头状瘤，前二者差异有显著性（ＰＭ０．０５），后二者差异无显著性（Ｐ＞０．０５）。Ａｇ－ＮＯＲｓ值与≥５Ｃ细胞百分数间有相关性ｒ＝０．５１１（Ｐ＜０．０５）。提示Ａｇ－ＮＯＲｓ值对喉良恶性病变有一定的鉴别价值，并在反映细胞增殖活性方面较ＤＮＡ倍体测定更加灵敏。     

局部晚期宫颈鳞癌流式细胞术分析与放射治疗预后的关系

 目的 研究局部晚期宫颈鳞癌流式细胞术分析结果（DNA倍体、SPF和PI）与放疗预后的关系，探讨它们在预测宫颈癌放疗效果中的价值。方法 68例ⅢB期宫颈鳞癌病人在放疗前钳取宫颈癌组织，制备成单细胞悬液，采用流式细胞术检测癌细胞DNA倍体、S期比例（SPF）及增殖指数（PI），分析他们及其他临床参数与患者根治性放疗后疾病复发和生存的关系。结果 在68例标本中，异倍体检出率为47．1％（32／68），SPF和PI值为分别为（7．49±2．91）和（12．89±3．75）；在随访期间，全组宫颈癌患者复发率为44．1％（30／68）；二倍体组和异倍体组的5年无复发生存率分别为52．7％和35．2％（P〈0．（15），高SPF组和低SPF组5年无复发生存率分别为25．3％和56．6％（P〈0．05），高PI组和低PI组5年无复发生存率分别为37．5％和49．3％（P〈0．05）；经多因素分析显示，肿瘤大小和SPF是影响宫颈癌患者无复发生存的独立预后因素。结论 通过流式细胞术检测宫颈癌组织SPF可以预测局部晚期宫颈鳞癌放射治疗后的预后。     

Prognostic value of nucleolar organizer regions and DNA ploidy in gastric-cancer

The prognostic value of nucleolar organizer region (Ag-NOR) and ploidy status in 233 gastric cancer was determined. In addition, correlation of bromodeoxyuridine labeling indices (BrdU LIs) and Ag-NOR scores was studied. The proportion of Ag-NOR scores varied from 1.85 to 12.0 (mean, 3.6; S.D., 1.9). A significant correlation was found between Ag-NOR scores and BrdU LIs, tumor size, serosal invasion, and lymph node metastasis. By univariate analysis, Ag-NOR score was proved to be a significant prognostic indicator. However, Cox regression analysis revealed that Ag-NOR scores and DNA ploidy were not independent prognostic parameters. However, when the combination analysis of Ag-NOR scores and DNA ploidy was entered into the Cox model, this parameter emerged as an independent prognostic parameter. These studies indicate that Ag-NOR scores reflect proliferative activity of gastric cancer, and that the Ag-NOR scores may possibly be useful prognostic parameters, when combined with DNA ploidy.     

鼻咽癌新鲜肿瘤组织DNA倍体性与预后的关系

背景与目的:因肿瘤有生物学异质性,部分肿瘤的预后和TNM分期并不符合;寻找有效的生物学预后指标作为临床分期的补充,可为今后鼻咽癌的个体化治疗提供一个新的依据.本研究探讨初治鼻咽癌患者新鲜肿瘤组织细胞的DNA倍体性与疗效、预后的关系.方法:1999年1月至2000年2月,53例初治鼻咽癌患者进入本研究,其中单纯放疗32例,另21例患者于放疗第4周接受了一个疗程的PF方案化疗.患者治疗前均活检取新鲜肿瘤组织,用流式细胞仪进行DNA倍体检测.结果:53例患者中,二倍体32例(60.4%),异倍体21例(39.6%).不同倍体组患者的年龄、性别、临床分期、N分期、化疗与否的差异无统计学意义(P=0.695、0.657、0.088、0.972和0.335).全组患者中位随访时间73个月(12～84个月).全组5年总生存率65.61%,其中二倍体组为80.92%,异倍体组为42.86%(P=0.002);5年无远处转移生存率二倍体组为84.26%,异倍体组为44.53%(P=0.003);5年无复发生存率二倍体组为92.59%,异倍体组为72.65%(P=0.118).单因素分析结果显示,临床分期是无复发生存率的影响因素,DNA倍体性、临床分期和T分期是总生存率和无远处转移生存率的影响因素.多因素分析结果显示,与总生存率相关的独立预后因素为DNA倍体性(P=0.020)和临床分期(P=0.007),与无转移生存率相关的预后因素亦为DNA倍体性(P=0.017)和临床分期(P=0.011).结论:采用流式细胞术检测新鲜组织细胞的DNA倍体性和临床分期一样可以预测鼻咽癌患者的预后;DNA异倍体患者比二倍体患者更容易出现远处转移而导致治疗失败.     

Measurement of bromodeoxyuridine labeling index, Ki-67 score and Ag-NOR count in breast carcinomas. Comparison with DNA ploidy.

Nuclear DNA content and three cell proliferation markers, the bromodeoxyuridine (BrdUrd) labeling index, Ki-67 score and Ag-nucleolar organizer region (Ag-NOR) counts, were measured in 21 cases of breast carcinoma, and then relationships among them were investigated. The BrdUrd labeling indices and Ki-67 scores were strongly correlated with one another (r = 0.90, p less than 0.001). They were also slightly correlated with the Ag-NOR counts (p less than 0.05). Although growth activity varied greatly from case to case, even within the same histologic grade in breast carcinoma, high histologic grade carcinomas were frequently associated with high values of both the BrdUrd labeling index and Ki-67 score, as compared with low histologic grade ones. The DNA ploidy level (DNA index) was independent of these cell proliferation markers. In the present series, 17 of 21 cases (81%) were aneuploid. All cases without DNA aneuploidy fell into a group of low histologic grade, while 7 of 11 low grade tumors were aneuploid. All three cell proliferation markers were low in 2 of the 4 cases without DNA aneuploidy. It may be possible to identify cases with favorable prognosis in a group of low grade carcinomas on the grounds of diploid DNA content and low values of these markers.     

Prognostic value of DNA ploidy in 653 Japanese women with node-negative breast cancer

Background. The prognostic value of DNA ploidy has been extensively studied in breast cancer; however, the results remain controversial. Flow cytometry (FCM) studies of DNA ploidy on frozen sections have not yet been performed in a large series of Japanese women with node-negative breast cancer. Methods. An FCM analysis of DNA ploidy was performed on frozen sections of node-negative breast cancer from 653 Japanese women, with a mean follow-up duration of 46 months. Results. Three hundred and twenty-four (49.6%) patients showed a diploid tumor, while 329 (50.4%) showed an aneuploid tumor. There was a significant correlation between DNA ploidy and estrogen receptor (ER) status. Patients with an aneuploid tumor had a significantly worse outcome in terms of both disease-free survival (DFS; P = 0.0116) and overall survival (OS; P = 0.0492) than those with a diploid tumor, while the same effect, in terms of DFS, was also seen in ER-positive breast cancer. Multivariate analyses indicated DNA ploidy to be an independent prognostic factor for DFS, while DNA ploidy and tumor size were found to be independent prognostic factors for OS. DNA ploidy was also shown to be an independent prognostic factor for DFS in ER-positive breast cancer. Conclusion. Our findings demonstrated DNA ploidy, based on FCM, to have an important prognostic value in Japanese women with node-negative breast cancer. Received: March 23, 2001 / Accepted: June 6, 2001     

Prognostic significance of the DNA content of renal carcinoma

DNA analysis was performed on fresh frozen samples of the primary tumor in 32 patients with renal carcinoma (13 with apparently localized disease and 19 with metastases at presentation). A comparison of ploidy with staging and standard histologic variables was performed. None of the patients who presented without metastases died of disease during the follow-up period. Eleven of 13 patients of this group had a diploid/near diploid pattern, and metastases developed in only one patient. Patients with metastatic disease and a diploid/near diploid DNA content had a significantly better survival rate than those with aneuploid primary tumors. Statistical analysis showed that grade and ploidy contributed significant but independent prognostic information. We concluded that DNA content is a useful prognostic factor in renal carcinoma.     

The clinical significance of nuclear DNA ploidy pattern in 184 patients with pheochromocytoma.

Flow cytometric nuclear DNA analysis was performed on paraffin-embedded tissue samples taken from 184 patients with pheochromocytoma and paraganglioma treated between 1960 and 1987. The Hedley technique was used for measurement of nuclear DNA content. Thirty-five percent of the tumors were DNA diploid, 33% showed a DNA tetraploid pattern, and 32% had DNA aneuploid pattern. Familial pheochromocytoma and associated endocrine or neoplastic disorders were more common among patients with DNA nondiploid tumors. Eighty-four percent of the tumors that invaded blood vessels and all patients with regional or distant metastases had tumors classified as DNA tetraploid or DNA aneuploid. Of 22 patients who had disease progression, 21 (95%) had tumors with abnormal DNA ploidy pattern (P less than 0.001). All 12 patients who died of cancer-related disease had abnormal DNA ploidy; none of the patients with DNA diploid tumor (n = 64) have died of pheochromocytoma (P less than 0.01). These results suggest that nuclear DNA ploidy pattern is an important and independent prognostic variable for patients with pheochromocytoma and paraganglioma.     

Carcinoma of the anal canal and flow cytometric DNA analysis

Using flow cytometric DNA analysis of paraffin embedded tissue, DNA histograms were successfully obtained from the anal cancers of 117 patients. DNA diploid patterns were given by 82 cancers (70%) and DNA non-diploid patterns by 35 cancers (30%): 15 DNA aneuploid, 20 DNA tetraploid. Well differentiated squamous cell cancers were mainly DNA diploid, while a larger proportion of poorly differentiated and small cell cancers were DNA non-diploid. The large majority of stage A cancers were DNA diploid. A greater proportion of tumours that had invaded through the anal sphincter or had lymph node metastases or distant spread were DNA non-diploid. Prognosis was slightly poorer for patients with DNA non-diploid cancers when compared to patients with DNA diploid tumours (P = 0.08) and significantly poorer for individuals with DNA aneuploid anal cancers (P = 0.037). However, in a multivariate analysis model, the DNA ploidy pattern of an anal cancer was not of independent prognostic significance alongside tumour histology and tumour stage.     

Prognostic value of S-phase fraction and DNA ploidy studied with in vivo administration of bromodeoxyuridine on human gastric cancers

The authors studied the prognostic values of DNA ploidy pattern and proliferative activity with in vivo administration of bromodeoxyuridine in human gastric cancers. Fresh specimens surgically removed from 117 patients with gastric cancer were investigated by flow cytometric study using a monoclonal antibody to bromodeoxyuridine. DNA ploidy patterns were classified into four types according to the bivariate BrdUrd/DNA distribution: D1, tumors with single diploid population; D2, tumors which showed mosaic of diploid and aneuploid population; A1, tumors with single aneuploid population; and A2, several aneuploid populations without diploid population. The numbers of cases of each ploidy pattern were as follows: D1, 36 cases (30.8%); D2, 38 cases (32.5%); A1, 15 cases (12.8%); and A2, 27 cases (23.1%). DNA ploidy pattern and S-phase fraction (SPF) showed no relation with clinicopathologic findings, except for type A2. In type A2, lymph node metastasis and lymphatic vessel invasion were observed more often than type D1. The SPF calculated from the bivariate BrdUrd/DNA distribution was higher in aneuploidy (D2, A1, and A2) than in diploidy (D1) (P less than 0.01). Also, A2 exhibited a higher SPF than A1 (P less than 0.01). Furthermore, SPF correlated with DNA index significantly (P less than 0.01). Patients who showed aneuploid tumors, DNA ploidy type A2, or SPF of more than 10% survived 3 years less than those with diploid tumors, DNA ploidy type D1, or SPF of less than 10%, respectively (P less than 0.05). By analyzing with the Cox's proportional hazard's model, it is revealed that DNA ploidy and SPF are one of the independent factors of prognostic significance. The results indicated that the patients with aneuploid tumors or highly proliferative tumors had a poor prognosis and that DNA ploidy pattern and SPF were useful prognostic factors for gastric cancers.     

Argyrophilic nucleolar organizer regions in breast-carcinoma - correlation with clinicopathological and biological variables and prognostic value

We investigated the relationship of argyrophilic nucleolar organizer regions (Ag-NOR) with clinico-pathologic and biologic variables, and its prognostic value in 127 patients with invasive breast carcinoma, Ag-NOR count was found to be significantly correlated with these variables, including DNA ploidy and c-erbB-2 expression. Although Ag-NOR count was a significant prognostic factor in overall and disease-free survival on univariate analysis, it did not appear to be an independent prognostic factor on multivariate analysis, in which status was introduced into the model. In patients in whom regional lymph node dissection has not been performed, however, Ag-NOR count may provide important prognostic information, especially when combined with c-erbB-2 expression.