Heterogeneity of DNA ploidy pattern in carcinoma of the gallbladder: primary and metastatic sites

There are few detailed reports on the heterogeneity of the nuclear DNA ploidy pattern in carcinoma of the gallbladder. We studied twelve autopsied cases who died of extended gallbladder carcinoma. Multiple samples were taken from the primary site (Pri), from direct invasion of the liver (Hinf), from hematogenous metastasis to the liver (H), from lymphatic metastasis (LN) and from peritoneal dissemination (P). The DNA ploidy pattern was investigated by image cytometry. Heterogeneity of the DNA ploidy pattern in Pri, Hinf, H, LN and P was found in 7/11, 2/10, 5/10, 2/6 and 3/6 cases, respectively. Aneuploidy was more frequently found in Hinf than at the Pri. The DNA index of Hinf was significantly higher than that of Pri. Several stemlines, with different quantities of DNA, were found in Pri. Most of these stemlines were also observed in other sites. These facts may suggest that polyclonal cancer cells rather than one cancer cell or monoclonal cancer cells of a Pri metastasize or infiltrate, and that various polyclonal cancer cells proliferate to different degrees under different circumstances.     

Fine needle aspiration of basal cell adenocarcinoma of the parotid gland. Report of a case with assessment of DNA ploidy in aspirates and tissue sections by image analysis

BACKGROUND: Basal cell adenocarcinoma of the parotid gland is a low grade malignant neoplasm. It has cytologic features of basal cell adenoma and a histologically infiltrative growth pattern of malignant tumors with perineural and vascular invasion. CASE: Fine needle aspiration biopsy findings of basal cell adenocarcinoma of the parotid gland in a 77-year-old male were supplemented by DNA ploidy analysis. CONCLUSION: No single cytologic feature was found to unequivocally distinguish this lesion from basal cell adenoma and/or solid variant of adenoid cystic carcinoma. Therefore, for diagnostic purposes, we grouped all three lesions under the term basal cell tumor. Evaluation of DNA content of tumor cells revealed diploid histograms in both cytologic material and paraffin-embedded tissue. Infiltrative tumor nests, the histologic basis for differentiating basal cell adenocarcinoma from adenoma, showed the same diploid pattern. Though DNA quantitation may not discriminate basal cell adenoma from basal cell adenocarcinoma, it may prove useful in separating them from adenoid cystic carcinoma, which is considered to be a tumor with high malignant potential.     

Adrenal pheochromocytoma mimicking small cell carcinoma on fine needle aspiration biopsy. A case report

BACKGROUND: Fine needle aspiration biopsy (FNAB) is a sensitive and specific technique in the diagnosis of adrenal tumors. However, in rare cases the cytomorphologic features may mimic small cell neoplasms. CASE: A 54-year-old male presented with a history of hypertension and left flank pain of recent onset. Abdominal computed tomography (CT) showed a 4-cm mass involving the left adrenal gland. Fine needle aspiration biopsy (FNAB) showed groups of small round cells with hyperchromatic nuclei. The findings were highly suspicious for metastatic small cell carcinoma. Subsequently it was learned that the mass had been noted three years earlier on CT studies but had grown from 2.5 to 4 cm. A chest radiograph was unremarkable. The clinical findings were more in keeping with a primary adrenal tumor. Immunohistochemical staining done retrospectively on the cell block showed positive reactivity for chromogranin and neuron-specific enolase. These findings, correlated with the clinical features, were in keeping with a diagnosis of pheochromocytoma. Left adrenalectomy revealed a pheochromocytoma. CONCLUSION: Adrenal pheochromocytoma should be included in the differential diagnosis of small round cell neoplasms seen on FNAB of the adrenal gland. Immunohistochemistry and clinical findings are helpful in reaching the correct diagnosis.     

Metastasis and invasion of hepatocellular carcinoma mimicking a right adrenal tumor

A 60-year-old man presented with a large right adrenal mass. Adrenal primary carcinoma invading the liver and retrohepatic inferior vena cava was suspected after preoperative imagings, which included ultrasonography, computed tomography, selective hepatic and adrenal angiography, and magnetic resonance imaging. An en bloc resection of the right kidney, right adrenal gland, posterior hepatic segment, and laterodorsal of the vena cava was performed using an active veno-venous bypass. The defect of the inferior vena cava was closed using a 6 x 10 cm patch of horse pericardium. The cut surface of the resected specimens revealed a smaller necrotic intrahepatic tumor as well as a large extrahepatic tumor which involved the right adrenal gland and extended continuously to the liver, mimicking an adrenal tumor. As the histological features of the two tumors disclosed the same moderately differentiated hepatocellular carcinoma with a trabecular or pseudoglandular pattern, a huge mass of the right adrenal gland with invasion into the right lobe of the liver, which mimicked a primary adrenal tumor, was diagnosed as metastatic hepatocellular carcinoma from a primary hepatic tumor.     

Evidence of clonal divergence in colorectal carcinoma

BACKGROUND: The aim of this study was to investigate the generation of DNA ploidy diversity in different stages of colorectal carcinoma development. METHODS: DNA flow cytometry was performed on tissue samples from 20 colorectal adenomas, 38 colorectal carcinomas, 30 lymph node metastases, and 70 hematogenous metastases. RESULTS: DNA aneuploidy was detected in 30% of the adenomas, 82% of the primary colorectal tumors, 57% of the lymph node metastases, 92% of the liver metastases, and 100% of the other distant hematogenous metastases. Multiple DNA tumor stemlines were found in 10%, 39%, 29%, 24%, and 40%, respectively. Sixty-two percent of the DNA tumor stemlines detected in the lymph node or liver metastases were also present in the primary tumors. In primary carcinomas and lymph node metastases, the DNA index distribution had a bimodal shape with a minimum at the 1.2-1.4 region. In the hematogenous metastases, a higher percentage of hypertetraploid stemlines was found. CONCLUSIONS: The emergence of DNA aneuploidy as well as clonal divergence seems to take place during the transition from adenoma to carcinoma. The DNA aneuploid stemlines formed during this phase remain relatively stable over time, although ongoing clonal evolution at distant metastatic tumor sites cannot be completely ruled out.     

Comparison of fluorescence in situ hybridization with flow cytometry and static image analysis in ploidy analysis of paraffin-embedded prostate adenocarcinoma

Nuclear DNA ploidy has been shown to have an important prognostic association for patients with adenocarcinoma of the prostate. Flow cytometry and static image analysis are ploidy methods that have been used in prostate carcinoma. Fluorescence in situ hybridization (FISH) using chromosome-specific probes can be used to evaluate the ploidy of interphase nuclei. In this study FISH was compared with flow cytometry and static image analysis in determining ploidy in paraffin-embedded tissue from 34 prostatic adenocarcinomas. Ploidy status using FISH was determined by enumerating centromeres of two chromosomes (8 and 12) by use of directly-labeled alpha-satellite DNA probes in isolated whole nuclei obtained by the Hedley technique. All three methods identified 11 of 34 cases as diploid and 17 of 34 cases as nondiploid (82% concordance). Six cases were discordant; two cases had discrepant results by each method. Ploidy classification as determined by FISH had an 88% concordance with ploidy classification by either flow cytometry or static image analysis. In conclusion, FISH was found to be a sensitive method of ploidy analysis in isolated paraffin-embedded nuclei from prostate adenocarcinomas. When the chromosomes commonly involved in aneuploidy have been identified in prostate adenocarcinoma, FISH has the potential to provide greater sensitivity for aneuploidy detection compared with currently available methods.     

Serum human hepatocyte growth factor (hHGF) is elevated in patients with metastatic gastric carcinoma

BACKGROUND/AIMS: Hepatocyte growth factor (HGF) is a stromally derived protein growth factor that modulates epithelial cell proliferation and motility. HGF may therefore be involved in tumor progression. METHODOLOGY: We measured the immunoreactive (ir)-HGF concentration in the sera of 56 patients with human gastric carcinoma, using an enzyme-linked immunosorbent assay and evaluated its association with clinical and histopathologic factors. Clinical stages were classified in accordance with The General Rules for TNM Classification (International Union Against Cancer). RESULTS: The serum hHGF concentrations from patients with gastric carcinoma significantly increased with increasing pathologic tumor grades. Also, there were significantly higher concentrations in patients with nodal metastasis compared to patients without metastasis. The serum hHGF concentrations in patients with liver metastasis were significantly higher than in patients without liver metastasis. CONCLUSIONS: These data show that the serum hHGF concentration is elevated in gastric carcinoma patients with liver and nodal metastasis. In multivariate analysis, the serum hHGF concentration was found to be the most important independent factor in predicting overall survival. Thus, the current results suggest that HGF may be important in the progression and metastasis of gastric carcinoma.     

Remote quantitation server for quality assurance in DNA ploidy analysis

OBJECTIVE: To remotely test a stepwise framework of quality control measures, developed according to the 1997 European Society of Analytical Cellular Pathology consensus on diagnostic DNA image cytometry, in two series of measurements by means of the quantitation server EUROQUANT. STUDY DESIGN: In each of these series, 104 fine needle aspiration biopsies, imprints from breast cancer specimens and 28 rat liver imprints were measured twice at two different cytometry workstations. Further measurements on special rat liver specimens for evaluation of the stability of the machinery and of the preparation process were done at both workstations. Afterwards the measurement data from both machines were transferred to the quantitation server and analyzed automatically. Beside the classical DNA histograms, a set of further evaluations was performed to detect optical errors as well as inhomogeneities in the measurements. Running values for mean integrated optical density values, mean corrective factors and mean coefficients of variations of corrective factors were computed to control the stability of the entire methodology over time. RESULTS: The study demonstrated the preconditions and outcome of server-based quality control in DNA ploidy analysis. The results show that such a remote analysis is feasible and comparable to local DNA ploidy analysis. It is also demonstrated how quality control tests reflect the process performance of ploidy analysis at its different levels. CONCLUSION: Quality control must be an inherent part of the daily routine to allow a reliable diagnostic interpretation and ensure steadily high product quality.     

Ultrasonographic study of adrenal gland tumors (report of 5 cases)

The authors study the ultrasound signs of the adrenal gland based on 5 different types of operated adrenal tumours (a lipoma, an adrenal cortical adenoma, a cyst, a corticoadrenaloma, an adrenal metastasis from a renal cell carcinoma). The ultrasonographic morphological criteria of the normal adrenal gland and each type of tumour are recalled and are illustrated by clinical cases. The value of ultrasonography in the investigation of this organ was evaluated in comparison with other imaging techniques. It constitutes a good first-line diagnostic guiding examination, but needs to be completed by other investigations (computed tomography, magnetic resonance imaging, etc.). Ultrasonography can also reveal asymptomatic adrenal tumours ("adrenal incidentalomas") in which the therapeutic decision (surveillance or surgery) must be based on criteria of size and appearance after a complete laboratory and morphological assessment.     

Cytomorphologic and DNA cytometric features of hepatocellular carcinoma in fine needle aspirates

OBJECTIVE: To classify hepatocellular carcinoma according to DNA ploidy patterns and to evaluate distinct cytomorphologic features of hepatocellular carcinoma that correlate with DNA ploidy patterns. STUDY DESIGN: Fine needle aspiration smears of 36 histologically proven hepatocellular carcinomas were performed for DNA measurement by image analysis after Feulgen restaining of the specimens. Nuclear features-prominent nucleoli, nuclear cleavage, nuclear area and nuclear/cytoplasmic ratio-were correlated with the DNA ploidy patterns. RESULTS: Of the 36 cases, 14 were either diploid (n = 7) or polyploid (n = 7), 19 tumors had a single aneuploid stemline, 2 cases had multiple stemlines, and 1 case had no discernible stemline. A preponderance of prominent nucleoli was seen in 7/7 diploid tumors (2c), 6/7 polyploid tumors (4c, 8c) and 6/8 aneuploid tumors (> 4c). Conspicuous nuclear cleavage in a high number of tumor cells was present substantially in tumors with large nuclear areas (4c, > 4c). CONCLUSION: Most hepatocellular carcinoma studied had a distinct stemline so that the tumors could be designated DNA diploid, polyploid or aneuploid. The prevalence of prominent nucleoli and nuclear cleavage was a distinguishing cytologic feature that could predict DNA ploidy patterns. No special association of the nuclear/cytoplasmic ratio with any of the ploidy groups was noted.     

Effort and achievement in national family planning programmes

Cell proliferation of 174 specimens obtained from the primary gastric cancers using endoscopic biopsy was investigated by immunohistochemical analysis with the monoclonal antibody PC10, which recognizes a proliferating cell nuclear antigen (PCNA) in formalin-fixed and paraffin-embedded material. All the examined samples showed nuclear staining for PCNA in cancer cells. The investigation was to test the correlation between PCNA labeling and lymph node metastasis. DNA aneuploidy was often encountered in tumors with nodal involvement and lymphatic invasion. The logistic regression analysis identified PCNA labeling rates (LRs), tumor size, and macroscopic type as independent significant factors for lymph node metastasis. When the PCNA LRs and clinicopathologic parameters were entered into the Cox regression analysis, PCNA LRs and DNA ploidy emerged as independent significant prognostic factors. In addition, combination assay of PCNA LRs and DNA ploidy yielded a powerful prognostic indication for patients with gastric cancer.     

Severe occlusive carotid artery disease: hemodynamic assessment by MR perfusion imaging in symptomatic patients

BACKGROUND: The purpose of this study was to assess the capability of magnetic resonance imaging (MRI) and cytology and flow cytometric (FCM) deoxyribonucleic acid (DNA) analysis in fine-needle aspiration biopsy (FNAB)-derived materials for diagnosing malignancy of the parotid lesions and the efficacy of FCM analysis in FNAB. METHODS: Magnetic resonance imaging findings and FCM results (ploidy and S + G2 + M phases [S + G2M] fraction) and cytology in FNAB-derived materials in 26 patients with 26 parotid lesions (12 benign lesions, 14 malignancies) were assessed for predicting malignancy. Flow cytometric results in aspirates were compared with those in surgically resected tissues. RESULTS: When a single predictor was used, cytology (92% accuracy) was most accurate for malignancy, followed by ill-defined margin (88% accuracy) and aneuploidy (88% accuracy). The combination of FCM and cytology raised the rate of sufficient materials from 92% to 100% and accuracy from 92% to 96% compared with cytology alone. The same highest accuracy (96%) was obtained with the combination of the ill-defined margin or other findings such as cytology, aneuploidy, or a high (S + G2M) fraction (6% <). Deoxyribonucleic acid ploidy in the FNAB showed full agreement with that in the surgical specimens. Receiver operating characteristic curves showed that the diagnosis of malignancy with (S + G2M) fraction in FNAB was superior to that in surgical specimens, but no significant difference was noted. CONCLUSIONS: A combination of MRI findings, cytology, and FCM results is optimal for diagnosing malignancies of the parotid lesions, and FNAB may replace the surgical specimens in FCM analysis.     

Fine needle aspiration biopsy of the liver. Principal diagnostic challenges

Fine needle aspiration biopsy (FNAB) is the diagnostic procedure of choice for evaluation of liver lesions. Although primarily applied to malignant disease, it is also used in the evaluation of benign conditions. Improvements in imaging techniques and advances in cytologic interpretations, as well as production of new biopsy needles, have all contributed to the rapid increase in radiologically guided liver FNAB.     

Importance of hepatic first-pass removal in metastasis of colon carcinoma cells

BACKGROUND/AIMS: It might be thought that colon carcinoma tends to metastasize to the liver because tumor cells leaving the primary colon tumor pass initially through the liver. Therefore we elucidated the kinetics of tumor cells in the body in order to understand the effect of the location of the liver on hepatic metastasis, that is to examine the hepatic first-pass effect of tumor cells. METHODS: Based on a physiological kinetic model, we examined quantitatively the hepatic metastasis and hepatic distribution of KM12-H1X cells administered by different routes. RESULTS: Both the number and incidence of colonies of hepatic metastasis were much greater after intrasplenic injection than after intravenous injection. The distribution of radioactivity to the liver after intrasplenic injection of [3H] thymidine-labeled cells was also much higher than that after intravenous injection. The number of colonies of hepatic metastasis correlated well with the area under the curve of the distributed amount of the tumor cells in the liver, regardless of the injection route; the correlation line was identical for each injection route. CONCLUSIONS: These results suggest that the hepatic first-pass effect is an important factor for the hepatic metastasis and that the cumulative number of tumor cells distributed in the liver is a determining factor for the degree of metastasis. Mathematical analysis based on a physiological model also suggests that hepatic metastasis depends on hepatic first-pass trapping of tumor cells.     

Operative management of adrenal metastases from lung carcinoma

Most surgeons consider patients with solitary adrenal metastasis from a primary lung carcinoma incurable and avoid excision of both the adrenal and primary lung tumors. However, several cases of successful surgical management of these patients recently have been reported. We reviewed 12 surgically treated patients with isolated adrenal and lung disease and identified 2 survivors of greater than fifteen years (17%) and 4 additional patients who are still alive following combined resection (34%). This survival rate, albeit in a selected population, represents an improvement over the natural history of nine months' survival. We suggest that if after six to twelve months of following patients with lung cancer and isolated adrenal metastasis no other evidence of spread of disease is evident, the tumor biology may be favorable and resection of both adrenal and lung lesions is reasonable.     

Predictors of survival for prostate carcinoma patients treated with salvage radical prostatectomy after radiation therapy

BACKGROUND: Salvage radical prostatectomy is a treatment option for patients with recurrent cancer following radiation therapy. This study was conducted to identify predictors of survival for patients treated with salvage radical prostatectomy. METHODS: The authors studied 86 prostate carcinoma patients who underwent salvage radical prostatectomy for locally persistent or recurrent prostate carcinoma at Mayo Clinic between 1967 and 1996. The mean interval from radiation therapy to biopsy-proven recurrence was 3.7 years (range, 6 months to 17 years). Patient age at surgery ranged from 51 to 78 years (median, 66 years). The mean follow-up after surgery was 5.8 years (range, 1.0-15.2 years). Cox proportional hazards models were used to identify clinical and pathologic factors associated with distant metastasis free survival and cancer specific survival. RESULTS: Actuarial distant metastasis free survival, cancer specific survival, and overall survival were 83%, 91%, and 85% at 5 years and 69%, 64%, and 54% at 10 years, respectively. In multivariate analysis, radical prostatectomy Gleason score and DNA ploidy were independent predictors of distant metastasis free survival and cancer specific survival. CONCLUSIONS: Postirradiation Gleason score and DNA ploidy were highly predictive of the clinical outcomes of patients treated by salvage radical prostatectomy after radiation therapy.     

The role of parathyroid hormone in the pathogenesis, prevention and treatment of postmenopausal osteoporosis

BACKGROUND: Nuclear deoxyribonucleic acid (DNA) content is a prognostic factor in several tumors, and decisions regarding treatment have been made using this parameter. Nevertheless, there is no agreement in head and neck cancer. The purpose of the present study was to ascertain whether tumor DNA content correlated with prognosis in cases of primary squamous cell carcinoma (SCC) of the oral cavity and tongue base. METHODS: A retrospective study of formalin-fixed, paraffin-embedded tissue from patients with histologically confirmed SCC of the oral cavity and tongue base was performed using flow cytometry. Tumor DNA content was studied in 109 sets of specimens from previously untreated patients. All of them underwent surgical resection at the University "Hospital de La Princesa" between 1982 and 1992. Clinical parameters (age, sex, site of primary tumor, clinical stage, adjuvant therapy received, and disease-free and overall survival) and histologic parameters (histopathologic stage, tumor differentiation, type of inflammatory infiltration, presence of perineural invasion) were recorded in all cases. An exhaustive statistical analysis was applied. RESULTS: Only the histograms of 93 patients were adequate for consideration. In flow cytometric analysis, DNA aneuploidy was observed in 51 tumors (55%). The proportion of aneuploid tumors was significantly higher in advanced-stage carcinomas (p < .05), tumors with perineural invasion (p < .05) and in men (p < .05). In the 24 patients with lymph node metastasis, the incidence of aneuploidy was 82% (19 of 24) (p < .05). The rate of metastasis and aneuploidy increased as the degree of differentiation decreased (p < .05 for both). Patients with aneuploid carcinomas in both early and advanced stages had shorter relapse-free and overall survival periods than did the patients with diploid tumors (p < .001 for both). A Cox regression analysis demonstrated that ploidy was the single most important prognostic factor in determining relapse and death (p < .001 for both). CONCLUSIONS: The results indicate that tumor DNA analysis by flow cytometry appears to be useful as a supplement to clinical and histologic evaluation in predicting the tendency of SCC of the oral cavity and tongue base to metastasize to regional lymph nodes and to predict the outcome of the disease.     

The invasion-MTT assay as a predictor of liver metastasis using human gastrointestinal carcinomas transplanted in nude mice

The invasion-3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay, which evaluates invasive potential into the reconstituted basement membrane, Matrigel, was performed on 49 human gastrointestinal carcinomas transplanted in nude mice. There were 19 colorectal carcinomas, 10 pancreatic carcinomas, 10 gastric carcinomas, 8 esophageal carcinomas, and 2 bile duct carcinomas. The percent invasion (PI) value of each tumor by the invasion-MTT assay expresses the invasive rate of tumor cells into the Matrigel as a percentage. There were no significant differences in correlations between the PI values and primary tumor site, clinicopathological findings, tumor doubling time, or DNA index; however, the PI values of primary tumors and lymph nodes with liver metastases were significantly higher than those of primary tumors without liver metastasis (P < 0.05). Furthermore, the primary tumors with synchronous (P < 0.05) or asynchronous (P < 0.01) liver metastases showed significantly higher PI values compared with the primary tumors without liver metastases. These results suggest that PI is not only an independent factor to predict liver metastasis, but it also correlates closely with liver metastasis. Thus, the invasion-MTT assay for primary tumors might be clinically useful to predict liver metastasis in patients following surgery for gastrointestinal carcinomas.     

Functional role of CD95 ligand in concanavalin A-induced intestinal intraepithelial lymphocyte cytotoxicity

OBJECTIVE: To update the analysis of technical and biologic factors related to hepatic resection for colorectal metastasis in a large single-institution series to identify important prognostic indicators and patterns of failure. SUMMARY BACKGROUND DATA: Surgical therapy for colorectal carcinoma metastatic to the liver is the only potentially curable treatment. Careful patient selection of those with resectable liver-only metastatic disease is crucial to the success of surgical therapy. METHODS: Two hundred forty-four consecutive patients undergoing curative hepatic resection for metastatic colorectal carcinoma were analyzed retrospectively. Variables examined included sex, stage of primary lesion, size of liver lesion(s), number of lesions, disease-free interval, ploidy, differentiation, preoperative carcinoembryonic antigen level, and operative factors such as resection margin, use of cryotherapy, intraoperative ultrasound, and blood loss. RESULTS: Surgical margin, number of lesions, and carcinoembryonic antigen (CEA) levels significantly control prognosis. Patients with only one or two liver lesions, a 1-cm surgical margin, and low CEA levels have a 5-year disease-free survival rate of more than 30%. Disease-free interval, original stage, bilobar involvement, size of metastasis, differentiation, and ploidy were not significant predictors of recurrence. The pattern of failure correlates with surgical margin. Routine use of intraoperative ultrasound resulted in an increased incidence of negative surgical margin during the period examined. CONCLUSIONS: Surgical resection or cryotherapy of hepatic metastasis from colorectal cancer is safe and curable in appropriately selected patients. Biologic factors, such as number of lesions and carcinoembryonic antigen levels, determine potential curability, and surgical margin governs the patterns of failure and outcome in potentially curable patients. Optimization of selection criteria and surgical resection margins will improve outcome.