Quantitative cytochemical detection of malignant and potentially malignant cells in the colon

It was found to be possible to distinguish malignant cells from normal cells by using an oxygen-sensitive tetrazolium salt (neotetrazolium) for the histochemical demonstration of glucose-6-phosphate dehydrogenase activity in cryostat sections of human colon. We have studied 12 cases of established adenocarcinoma of the colon in addition to 4 of ulcerative colitis and 4 of adenomatous polyposis (polyposis coli). In a nitrogen atmosphere the activities of malignant and normal cells were similar. However, after incubation in an atmosphere of pure oxygen, only malignant cells gave a positive reaction after 5 min. Three of the four cases of adenomatous polyposis gave a positive reaction for glucose-6-phosphate dehydrogenase activity in oxygen in a manner similar to that of specimens with severe dysplasia. In general, positive foci were histologically indistinguishable from the neighboring tubuli. However, foci of severely dysplastic epithelium usually showed a positive reaction. All three patients eventually developed clear-cut severe dysplasia. The other patient, who showed a negative reaction in oxygen, was diagnosed after 3 years as not suffering from dysplasia. All cases of ulcerative colitis gave a reaction in oxygen comparable with that of normal cells. Therefore, the areas with a positive reaction are considered to be either in the process of malignant transformation or malignant. An explanation for the oxygen insensitivity of cancer cells appeared to be a decrease in the activity of superoxide dismutase (EC 1.15.1.1), as addition of exogenous superoxide dismutase to malignant cells caused a normal reaction. We wish to suggest that this test in combination with the routine histology may be exploited for the diagnosis of polyps in premalignant conditions.     

Diagnostic p53 immunostaining of endobiliary brush cytology: preoperative cytology compared with the surgical specimen.

BACKGROUND: Endobiliary brush cytology is important in the distinction of malignant and benign causes of extrahepatic bile duct obstruction. The additional diagnostic value of p53 immunostaining on these cytology specimens was assessed. METHODS: All patients with extrahepatic bile duct obstruction who underwent endoscopic retrograde cholangiopancreatography (ERCP) with endobiliary brush cytology and subsequent surgery at the Academic Medical Center in Amsterdam during a 3-year period were studied. p53 Immunocytology was compared with the corresponding conventional light microscopic cytology and p53 immunostaining of the subsequent surgical specimen. RESULTS: Fifty-three patients with the following diagnoses were included: pancreatic carcinoma (23), bile duct carcinoma (15), ampullary carcinoma (5), lymph node metastases (2), carcinoma of unknown origin (4), chronic pancreatitis (3), and primary sclerosing cholangitis (1). Fifty-one percent of the carcinomas showed positive p53 immunostaining; all four surgical specimens without carcinoma were negative. The sensitivities of conventional light microscopic cytology, p53 immunocytology, and both tests combined were 29%, 24%, and 43%, respectively. These sensitivities were higher in cases of bile duct carcinoma (46%, 40%, and 66%) compared with cases of pancreatic carcinoma (13%, 9%, and 22%). Specificities of both tests were 100%. CONCLUSIONS: p53 Immunostaining on endobiliary brush cytology may be helpful in the diagnosis of malignant extrahepatic bile duct stenosis, especially in patients with bile duct carcinoma. Cancer (Cancer Cytopathol) Copyright 1999 American Cancer Society.     

Problems in the diagnosis of small cell carcinoma of the lungs by fiberoptic bronchoscopy

Fiberoptic bronchoscopy (FOB) with the aid of endoscopic biopsies and brush cytology is recognized as a valuable approach in the diagnosis of lung cancer. However, histologic classification of lung cancer based on tiny specimens obtained from FOB can be difficult. Correct identification of small cell carcinoma of the lung is especially important because its recognition usually precludes surgery. In a review of 770 patients who underwent FOB biopsies at The Mount Sinai Hospital, New York, in individuals with proven lung cancer 150 instances of small cell carcinoma were encountered. In four of these instances subsequent surgery, such as scalene node biopsy, mediastinoscopy, or thoracotomy, was performed because clinically and radiologically the tumors did not behave as small cell carcinomas. Pathologic examination of larger tissue samples from these neoplasms provided the following final diagnoses: bronchial carcinoid, adenocarcinoma, squamous cell carcinoma, and small cell carcinomas-combined type. Analysis of the FOB biopsies and brush cytology usually permit diagnosis of small cell carcinoma of the lung. However, in instances where the biologic behavior of a tumor casts doubt on the diagnosis of small cell carcinoma, further studies should be performed, including radionuclide scans, and bone marrow and other biopsies before denying the patient a chance of surgical cure.     

Diagnosis of Oral Cancers by Targeting VPAC Receptors: Preliminary Report

Introduction: Oral cancer is a major health problem. The study of exfoliative cytology material helps in the differentiation of premalignant and malignant alterations of oral lesions. The objective of this study was to assess the feasibility of detecting oral cancer by targeting genomic VPAC (combined vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase-activating peptide) receptors expressed on malignant oral cancer cells. Patients & Methods: All patients with suspected oral cavity cancers/lesions formed the study group. The samples from the oral cavity lesion or suspicious area were collected with a cytology brush. The harvested material was examined for malignant cells by 1. the standard PAP stain and 2. targeting the VPAC receptors on the cell surface using a fluorescent microscope. Similarly, malignant cells were identified from cells shed in oral gargles. Results: A total of 60 patients with oral lesions were included in the study. The histopathological diagnosis was squamous cell carcinoma in 30 of these. The VPAC receptor positivity both on the brush cytology staining as well oral gargle staining was more sensitive than the brush cytology PAP staining. The accuracy of the various techniques was as follows, brush cytology PAP staining at 86.67%, brush cytology VPAC staining at 91.67% and oral gargle VPAC staining at 95%. Conclusions: This preliminary study validates our belief that malignant cells in the saliva can be identified by targeting the VPAC receptors. The test is simple, easy, non-invasive and reliable in the detection of oral cancers.     

Clinical value of K-ras codon 12 analysis and endobiliary brush cytology for the diagnosis of malignant extrahepatic bile duct stenosis.

Extrahepatic biliary stenosis can be caused by benign and malignant disorders. In most cases, a tissue diagnosis is needed for optimal management of patients, but the sensitivity of biliary cytology for the diagnosis of a malignancy is relatively low. The additional diagnostic value of K-ras mutational analysis of endobiliary brush cytology was assessed. Endobiliary brush cytology specimens obtained during endoscopic retrograde cholangiopancreaticography were prospectively collected from 312 consecutive patients with extrahepatic biliary stenosis. The results of conventional light microscopic cytology and K-ras codon 12 mutational analysis were compared and evaluated in view of the final diagnosis made by histological examination of the stenotic lesion and/or patient follow-up. The sensitivities of cytology and mutational analysis to detect malignancy were 36 and 42%, respectively. When both tests were combined, the sensitivity increased to 62%. The specificity of cytology was 98%, and the specificity of the mutational analysis and of both tests combined was 89%. Positive predictive values for cytology, mutational analysis, and both tests combined were 98, 92, and 94%, whereas the corresponding negative predictive values were 34, 34, and 44%, respectively. The sensitivity of K-ras mutational analysis was 63% for pancreatic carcinomas compared to 27% for bile duct, gallbladder, and ampullary carcinomas. K-ras mutational analysis can be considered supplementary to conventional light microscopy of endobiliary brush cytology to diagnose patients with malignant extrahepatic biliary stenosis, particularly in the case of pancreatic cancer. The presence of a K-ras codon 12 mutation in endobiliary brush cytology per se supports a clinical suspicion of malignancy, even when the conventional cytology is negative or equivocal.     

Overexpression of p53 protein during pancreatitis.

Overexpression of p53 correlates with neoplasia in many cytological specimens. To test the specificity of overexpressed p53 as a tumour marker for the detection of pancreatic cancer, we analysed cytological specimens of pancreatic juice samples from patients with pancreatitis or pancreatic carcinoma (n = 42) for p53 protein overexpression. p53 protein overexpression was found in 59% of patients with pancreatitis and 67% of patients with pancreatic carcinoma. Thus, the assessment of p53 protein overexpression is not useful in the diagnosis of pancreatic cancer. Overexpressed p53 during pancreatitis appears to be wild-type p53. Overexpression of p53 may result from DNA damage occurring during chronic inflammation. It is well established that p53 can induce apoptosis upon DNA damage. Consequently, we found apoptotic cell death in five out of five tested cytological preparations from patients with pancreatitis as well as in one out of one pancreatic carcinoma specimen.     

Brush cytology in the diagnosis of colonic neoplasms

During a three-year period (1986-1988), 234 colonic brush specimens were received in the authors' laboratory. Nine samples (4%) were deemed unsatisfactory for evaluation because of inadequate cellularity and/or poor fixation. In 11 cases concomitant or follow-up histologic specimens were not available. The remaining 214 specimens included 82 malignant neoplasms, 88 neoplastic polyps (adenomas), and 44 nonneoplastic lesions. Sixty-seven (82%) of malignant neoplasms were correctly diagnosed by brush cytology. Three cases of adenoma with severe dysplasia or in situ carcinoma were diagnosed as adenocarcinoma by cytology. No false-positive diagnoses were made of nonneoplastic lesions. Brush cytology was found to be a more sensitive technique in the diagnosis of colon cancer than endoscopic biopsy (82% and 74% sensitivity, respectively). The combination of the two techniques increased the sensitivity to 90% and improved the overall accuracy of the test. Seventy-one (82%) of the colonic adenomas were correctly diagnosed by cytology. Brush cytology is a convenient, safe, and accurate technique which should be used concurrently with endoscopic biopsy or polypectomy.     

Case of pancreatic metastasis from colon cancer in which cell block using the Trefle® endoscopic scraper enables differential diagnosis from pancreatic cancer

Endoscopic transpapillary brush cytology and forceps biopsy during endoscopic retrograde cholangiopancreatology are generally used to obtain pathological evidence of biliary strictures. Recently, the new endoscopic scraper Trefle^® has been reported and demonstrated high cancer detectability in malignant biliary strictures. This device is used to scrape the stricture over the guidewire, and, in the original method, the tissue and/or cell samples obtained are subjected to histological and/or cytological analysis separately. However, discrimination of chunks of tissue is hampered by the opacity of the surrounding fluid. We have developed a cell block technique for the Trefle^® device without dividing obtained specimens into tissue and cellular components, which is the simplest method and enables immunohistochemical analysis. We present a case of obstructive jaundice diagnosed immunohistochemically as pancreatic metastasis from colon cancer using cell block sections obtained with the Trefle^® device, which procedure is as easy as conventional brush cytology.     

Immunohistochemical analysis of p53 and MIB-1 in tissue specimens obtained from endoscopic ultrasonography-guided fine needle aspiration biopsy for the diagnosis of solid pancreatic masses

Endoscopic ultrasonography-guided fine-needle aspiration biopsy (EUS-FNAB) has been shown to be a highly accurate technique for distinguishing benign from malignant pancreatic masses. In this study, we examined p53 immunohistochemical analysis in FNAB specimens obtained from solid pancreatic diseases, and prospectively evaluated clinical applications for the diagnosis of malignancy in combination routine histological examination. Tissue specimens obtained from 62 pancreatic masses (51 pancreatic cancers and 11 chronic pancreatitis) by EUS-FNAB were evaluated by routine histological examination and p53 immunostaining. The conventional EUS-FNA diagnostic test statistics for the pancreatic masses were as follows: 76% sensitivity, 91% specificity and 79% accuracy. p53 protein overexpression was observed in 67% patients with pancreatic cancer, but not in patients with chronic pancreatitis. If the diagnosis of malignancy was made using the combination of p53 protein overexpression and conventional histological examination, the diagnostic test statistics changed as follows: 90% sensitivity, 91% specificity and 92% accuracy. p53 immunostaining in combination with routine histological examination of EUS-FNAB may improve the diagnostic accuracy for pancreatic cancer.     

痰和纤维支气管镜刷片细胞学检查在肺癌诊断中的意义

目的探讨痰和纤支镜刷片细胞学检查在肺癌诊断中的意义。方法收集532例同时进行痰和纤支镜检查的肺癌病例,分析痰和纤支镜刷片细胞学检查的敏感性和分型的准确性,与纤支镜咬检组织学对比,评价细胞学在肺癌诊断中的意义。结果痰细胞学的敏感性为33．8％,纤支镜刷片细胞学的敏感性为58．7％,纤支镜咬检组织学的敏感性为39．1％。痰细胞学分型诊断与组织学的符合率鳞癌为95．7％,腺癌为87．2％,小细胞癌为100．0％,痰细胞学区分小细胞癌和非小细胞癌的准确性为100．0％。纤支镜刷片细胞学分型诊断与组织学的符合率鳞癌为93．3％,腺癌为85．4％,小细胞癌为95．5％,纤支镜刷片细胞学区分小细胞和非小细胞癌的准确性为98．0％。结论纤支镜刷片细胞学具有较高的敏感性,在肺癌的诊断中有重要的应用价值。痰细胞学敏感性较低,但可作为纤支镜检查阴性和不能耐受纤支镜检查的肺癌患者的补充检查手段。两者的细胞学分型具有较高的可信度。     

Loss of the Y chromosome is a frequent chromosomal imbalance in pancreatic cancer and allows differentiation to chronic pancreatitis

In a study for the identification of genomic alterations in pancreatic cancer, representational difference analysis was used and led to the isolation of 2 distinct fragments, deleted on the Y chromosome in the xenografted tumor DNA of a male patient with an adenocarcinoma of the pancreas. Loss of Y chromosomal material was further studied in 11 pancreatic cancer cell lines of male origin, using PCR amplification of 5 sequence tagged sites (STSs) distributed along the Y chromosome; 8/11 cell lines exhibited a complete loss of the Y chromosome and 3 had deletions. To examine the status of the Y chromosome in situ, interphase FISH analysis was performed on paraffin sections from pancreatic carcinoma (n=7) and chronic pancreatitis (n=7) tissues, and the loss of Y-chromosomal STS-markers was studied in 6 xenograft tumors obtained from male pancreatic cancer patients. This analysis revealed that a loss of the Y chromosome occurs in vivo in primary pancreatic tumor cells, whereas the Y chromosome was intact in chronic pancreatitis. Our data suggest that loss of Y is a frequent event occurring in male pancreatic tumors. Although there is no evidence for a functional implication of Y chromosome loss, it effectively differentiates between a malignant and a benign condition as e.g. chronic pancreatitis. Thus, this genetic alteration may be of diagnostic use.     

应用组织芯片技术研究人类胰腺癌中PSCA的表达

目的检测PSCA在胰腺癌中的表达情况，探讨PSCA在胰腺癌发病中所起的作用。方法用组织芯片技术构建包含78例导管腺癌，12例慢性胰腺炎病人，10例正常胰腺组织的100点阵的石蜡组织芯片。用免疫组化SP法检测该芯片中PSCA的表达，分析其与胰腺癌临床病理因素的关系。结果78例胰腺癌病人中，PSCA阳性表达率为79．5％（62／78），与正常组比较PSCA表达与胰腺癌显著相关（χ^2=15．81，P〈0．005），与慢性胰腺炎比较PSCA亦与胰腺癌显著相关（χ^2=11．33，P〈0．005）；PSCA表达与年龄、性别、组织分化程度及TNM分期无明显相关性。结论PSCA阳性表达与胰腺癌相关，可能与胰腺癌的发生发展有着密切关系，但与胰腺癌的临床病理特型无关。     

ErbB2 oncogene antibodies differentiate between the normal and diseased pancreas, and between chronic pancreatitis and pancreatic cancer

Histological differentiation between chronic pancreatitis and pancreatic cancer, especially in biopsy material, remains challenging and the frequent association of 'secondary' chronic pancreatitis (due to ductal obstruction) with pancreatic cancer causes additional diagnostic problems. Our study, using anti-ErbB2 antibodies from Santa Cruz and Dako in tissues from the normal pancreas, chronic pancreatitis and pancreatic cancer showed that these antibodies discriminate between primary chronic pancreatitis and 'secondary' chronic pancreatitis due to pancreatic cancer. Tissues from 28 pancreatic cancer patients, 15 chronic pancreatitis patients and 12 organ donors or early autopsy cases were subjected to immunohistochemical studies using polyclonal ErbB2 antibodies from Santa Cruz and Dako. The Santa Cruz antibody immunoreacted with islet cells in all tissues from the normal pancreas and pancreatic cancer but not in any chronic pancreatitis specimen. The Dako antibody showed a membrane staining of ductal and ductular cells only in chronic pancreatitis cases but in none of the normal or cancer specimens. Moreover, in chronic pancreatitis cases, ductular cells were stained with the Santa Cruz antibody only in the severe form, but not in the mild or moderate form of the disease. The utilized ErbB2 antibodies discriminate between the normal pancreas, chronic pancreatitis and pancreatic cancer. Hence, these antibodies seem to present an additional useful aid in the surgical pathology of pancreatic diseases.     

The glycolytic enzyme activity of the human cervix uteri.

The activity of the following enzymes was studied in normal, precancerous, and malignant biopsies from the human cervix uteri: hexokinase (HK), phosphofructokinase (PFK), pyruvate-kinase (PK), lactate dehydrogenase (LDH), and glucose-6-phosphate dehydrogenase (G-6-PDH). In precancerous conditions, i.e., dysplasia and carcinoma in situ without any signs of invasive carcinoma, only PK showed moderate but significant activity increases. A rise in enzyme activity in biopsies histologically classified as carcinoma in situ was found to signal the presence of invasive carcinoma in other parts of the cervix. In invasive carcinomas of the cervix, all the enzymes studied showed a two- to four-fold increase (p less than 0.01) as compared to the normal cervix. The present study failed to reveal significant differences between enzyme activities in biopsies from patients in Stage I, II, and III; no correlation could be established between enzyme activity and prognosis.     

顺序特异性引物法对胰腺癌K—ras基因点突变的研究

为研究简捷、特异、敏感的检测胰腺癌Ｋｒａｓ基因点突变的方法及其在胰腺疾病中定性诊断的价值,采用针对该基因点突变方式（ＣＧＴ、ＧＡＴ、ＧＴＴ）设计的顺序特异性引物（ＳＳＰ）,先后对胰腺癌石蜡包埋组织、冰冻新鲜组织、细针穿刺组织及胰液进行多聚酶链反应（ＰＣＲ）,扩增产物借助常规电泳和染色检测有无Ｋｒａｓ基因突变及突变方式。结果显示：胰腺癌石蜡包埋组织、冰冻新鲜组织、细针穿刺组织及胰液中Ｋｒａｓ基因点突变率分别为７４．２％、９５．１％、９１．４％及９４．１％,而所有被检测的慢性胰腺炎、胰岛素瘤、壶腹癌、胆管癌、十二指肠乳头癌及外伤胰腺的组织标本和胰液标本均无Ｋｒａｓ基因突变,无假阳性发生。研究表明：该检测法快速、简便、特异、敏感,具有临床实用性,可以作为鉴别胰腺肿块良、恶性和诊断胰腺癌的一种方法