Giant cell tumor of bone. A clinicopathologic and DNA flow cytometric analysis

Flow cytometric DNA analysis was performed on 60 cases of giant cell tumor of bone and the results were correlated with the clinicopathologic features. Tumors studied were from 31 men and 29 women whose ages ranged from 18 to 62 years (median, 29 years). The most common sites were the distal end of the femur and proximal end of the tibia, accounting for 75% of the lesions. Treatment consisted of resection in 29 patients (48%), curettage with bone chip packing in 15 patients (25%), or curettage with cement packing in 16 patients (27%). Ten patients (17%) had local relapse within 1 to 3 years, and two had lung metastases. Forty-two patients (70%) exhibited tumors with a diploid DNA content, 16 aneuploid (27%), and two tetraploid (3%). Six (37.5%) of the aneuploid patients had relapses: one of those had been treated by resection of the tumor and five by curettage. Of the remaining ten (62.5%) unrelapsed aneuploid patients, nine had been treated by resection of the tumor and one by curettage. Four of the 42 diploid patients (9.5%) had relapses; all had been treated by curettage of the tumor. The two tetraploid tumors were treated by resection and none relapsed. Histologic parameters did not correlate with relapse rate or DNA pattern. Although relapse was more common among aneuploid tumors, our study shows that this appears to be influenced by the treatment modality rather than the ploidy status. Based on this study the DNA analysis of giant cell tumor of bone has a limited utility for predicting the tumor's biologic behavior.     

Primary sarcomas of the kidney. A clinicopathologic and DNA flow cytometric study of 17 cases

Primary sarcomas of the kidney in adults are rare. In the handful of published reports of all soft tissue sarcomas, DNA ploidy has correlated with histologic grade and outcome. This report presents the clinicopathologic and flow cytometric features of 17 cases of primary renal sarcoma (seven men and ten women, ages 28-69 years). Presenting symptoms included abdominal and back pain and hematuria. Stages at diagnosis were I, in three patients; II, five patients; III, two patients; and IV, two patients. Eight tumors were leiomyosarcoma, two malignant fibrous histiocytoma, one hemangiopericytoma, one fibrosarcoma, and five unclassified. Tumors measured 5.5 to 23 cm, seven contained marked nuclear pleomorphism, seven were extensively necrotic, and mitotic rate was 1 to 33 per 10 high-power fields. Seven tumors showed aneuploidy and five were diploid. Thirteen patients were dead of disease after a mean of 23 months and two were alive with known metastases at 29 and 33 months, respectively. Ploidy pattern and outcome or time to death were not correlated, but aneuploidy correlated with histologic grade, marked nuclear pleomorphism (P less than 0.05), extensive necrosis (p less than 0.01), and high mitotic rate (0.05 less than P less than 0.10). The authors conclude that although DNA ploidy does correlate with histologic grade, for primary renal sarcomas, whose prognosis in this series was extremely poor, it does not correlate with outcome.     

Simultaneous carcinoma involving the endometrium and the ovary. A clinicopathologic, immunohistochemical, and DNA flow cytometric study of 18 cases

Eighteen carcinomas involving both the endometrium and the ovary were studied. Stage, size, bilaterality and pattern of ovarian involvement, histologic types and grades, presence of endometrial hyperplasia or ovarian endometriosis, myometrial, tubal, lymphatic and blood vessel invasion, and follow-up of the patients were all evaluated. Accordingly, the cases were classified as follows: Group A (nine cases), two separate primary tumors; and Group B (nine cases), uterine primaries with ovarian metastasis or ovarian primaries with uterine metastasis. Immunohistochemical stains (CAM 5.2, wide-spectrum keratin, vimentin, carcinoembryonic antigen (CEA), CA 12.5, CA 19.9) were performed in all cases. Flow cytometric determination of nuclear DNA was done in 13 (seven Group A and six Group B tumors). Of the nine cases with independent primary tumors, seven showed different immunohistochemical profiles in the ovarian and uterine tumors, whereas only four of the nine metastatic ones had similar staining characteristics. Five cases with independent primary tumors, but only one of the metastatic group, exhibited different aneuploid stemlines in the endometrial and ovarian tumors. The other seven (two independent and five metastatic) cases had similar DNA indexes in both tumors. Immunohistochemical and DNA flow cytometric study may be of some value for the distinction between metastatic and independent tumors, but differential diagnosis must presently rely largely upon conventional clinicopathologic criteria.     

Renal oncocytoma: long-term follow-up and flow cytometric DNA analysis

We report a retrospective study on the clinicopathologic features and flow cytometric DNA analysis of ten renal oncocytomas compared with a control group of ten randomly selected renal cell carcinomas. Among the oncocytoma patients, no recurrences or metastases were noted over an average follow-up of 6.7 years (range = 6 months to 16 years). Reproducible, high-quality DNA histograms were obtained on the paraffin-embedded specimens by using our modified flow cytometric procedure. One aneuploid (10%) and two hyperdiploid tumors (20%) were found in the oncocytoma group. There was no correlation between these abnormal DNA histographic patterns and survival or tumor stages. On the contrary, a good correlation was found between tumor grades and DNA ploidy in the controls. We conclude that renal oncocytoma is a clinically benign tumor, yet it may exhibit varying degrees of flow cytometric DNA abnormalities, which have no predictive value on survival and probably reflect the characteristics of oncocytes rather than its malignant potential.     

A flow cytometric analysis of DNA content in primary and metastatic lesions of esophageal squamous cell carcinoma.

BACKGROUND. DNA content of malignant tumors has been considered a significant prognostic factor, but intratumor variations in DNA content and differences in DNA content between primary and metastatic lesions have been found in various tumors. METHODS. DNA indexes of multiple samples obtained from different sites in each tumor were determined by flow cytometry (FCM) in 27 esophageal squamous cell carcinomas. RESULTS. Intratumor variation in DNA indexes in primary lesions was found in 10 (37%) of the 27 specimens. Of the rest, all DNA indexes were diploid in 5 and all identically aneuploid in 12 samples. A DNA index differing from that of the metastatic lesion was found in four (50%) of eight primary lesions; however, a component of the DNA index identical to that of metastatic lesion was found in seven (88%) of eight primary lesions. CONCLUSIONS. Recurrence after a "curative" operation was frequent in patients with a tumor that had an aneuploid DNA index, with or without a variation in DNA indexes. There was no recurrence in the five patients with tumors that had only a diploid DNA index. These results suggest that differences in DNA content between primary and metastatic lesions reflect intratumor variation in DNA content in the primary lesions. The presence of a tumor cell population with an aneuploid DNA content, even when the DNA content varied in the primary lesion, may indicate an aggressive clinical course in patients with esophageal squamous cell carcinoma.     

Cytogenetic and corresponding flow cytometric DNA analysis of renal cell neoplasms.

Results of flow cytometric DNA content and cytogenetic analyses of six neoplasms representing the spectrum of the morphologic subtypes of renal cell neoplasms are compared. Flow cytometric determinations of DNA diploidy and near diploidy in two renal cortical neoplasms correlated with modal chromosome numbers; however, in three of four neoplasms with a highly aneuploid DNA content, the modal chromosome numbers indicated diploidy or near-diploidy. Our data suggest that the short-term culture conditions used in cytogenetic analysis may favor growth of cells with diploid or near-diploid DNA content. This may explain the discrepancy frequently observed between results obtained using flow cytometry and karyotyping after short-term culture. We also observed (1) abnormalities in the short arm of chromosome 3 in all three nonpapillary neoplasms, (both clear and granular cell types); (2) aberrations in chromosomes 3 and 17 in the two papillary tumors; and (3) normal chromosome 3 in the presence of various random karyotypic anomalies including telomeric associations and a pseudo-diploid modal chromosome number in the only oncocytic neoplasm studied.     

DNA flow cytometric analysis and prognosis of axillary lymph node-negative breast carcinoma.

METHODS. The prognostic significance of flow cytometric analysis in patients with node-negative invasive breast carcinoma was evaluated in a retrospective series of 158 patients with a minimum follow-up study of 9 years. RESULTS. The ploidy status could be assessed in 147 specimens (93%), and the proliferative phase or S-phase fraction (SPF) could be assessed in 136 tumors (86%); 70 tumors (48%) were diploid, 49 tumors (33%) were aneuploid, and 28 tumors (19%) were tetraploid. Ploidy status and SPF were correlated significantly with tumor size, histologic grade, nuclear grade, and mitotic rate. By itself, ploidy was not a statistically significant prognostic factor, although all of the patients with multiploid and hypertetraploid tumors had recurrence of disease. The SPF was related significantly to recurrence of disease (P = 0.04). However, when multivariate analysis of various histopathologic variables was performed, SPF ceased to be a significant prognostic determinant, whereas peritumoral lymphovascular invasion was the most important variable. The combination of tumor size and flow cytometric parameters permitted stratification into three groups with different prognoses at the 9-year follow-up review (P less than 0.001). In the low-risk group (diploid tumors less than or equal to 2 cm in diameter with a low SPF or small tetraploid tumors), the recurrence rate was 12%. In the intermediate-risk group (diploid tumors greater than 2 cm in diameter with a low SPF or aneuploid tumors with a low SPF), the recurrence rate was 21%. In the high-risk group (diploid or aneuploid tumors with a high SPF or large tetraploid tumors), the recurrence rate was 49%. The high-risk group status remained a significant variable in the Cox proportional hazards multivariate analysis model. CONCLUSIONS. These results indicate that flow cytometry in breast carcinoma contributes useful but limited prognostic information and stress the importance of using multiple prognostic factors to improve prognostication and optimize patient management.     

Small cell carcinoma of the urinary bladder. A clinicopathologic analysis of 22 cases.

Small cell carcinoma of the urinary bladder is an uncommon tumor. The authors report the clinicopathologic findings in a series of 22 cases. Fifteen men and 7 women were studied; their ages ranged from 51 to 87 years (mean, 62.4 years). The most frequent presentation was hematuria (94.4%). At diagnosis, three patients had Stage B disease, six had Stage C, and ten had Stage D (unknown stage in three). Histologically, 6 were oat cell type tumors, 11 were of intermediate cell type, and 5 were of combined cell type. Immunohistochemical studies demonstrated positivity for neuron-specific enolase in ten of ten cases, cytokeratin in seven of ten cases, chromogranin in eight of nine cases, serotonin in seven of nine cases, and S-100 protein in four of ten cases. Neuroendocrine differentiation was seen in five of seven cases examined by electron microscopy. Treatment and follow-up data were available for 19 patients: 10 (52.6%) were dead of disease, 5 (26.3%) were alive and well, 3 (15.8%) were alive with disease, and 1 (5.3%) died of an unrelated cause. The 2-year survival rate was 50% for patients with Stage B, 25% for patients with Stage C, and 33% for patients with Stage D disease. Although overall survival was poor, some cases responded well to therapy. Based on the authors' experience, radical cystectomy with adjuvant chemotherapy appears to be the treatment of choice.     

DNA flow cytometric analysis of human nasopharyngeal carcinoma in nude mice

The tissue from a patient with nasopharyngeal carcinoma has been transferred to nude mice (BALB/c), and has successfully been growing through twenty passages. The tumors in the nude mice and primary human tumor have been examined for the cellular DNA content by FCM and also conventional pathological examination, chromosome analysis, and EBV test. The tumor take rate varied markedly in different passages with a mean value of approximately 70%, and showing a tendency to increase. The tumor doubling time within 6-12 weeks after transplantation of six tumors in 18 and 20 passages were 14.8 and 9.3 days respectively. However, the tumor volume at 12 weeks varied significantly, ranging from 438 to 1998 mm3. By FCM, it has been found that the values of DNA index were about the same in both primary tumor and the tumors in nude mice. The distribution of various phase cells in cell cycle was also about the same in both. In conclusion, the application of FCM to examine the cellular DNA content of the tumor in nude mice is a rapid and sensitive method, useful in the investigation on the stability of biological characteristics of human NPC in nude mice and in further studies on the effects by various therapeutic methods.     

Flow cytometric DNA analysis of hepatocellular carcinoma.

The prognostic value of nuclear DNA content was studied retrospectively using flow cytometry in 203 cases of resected hepatocellular carcinoma. The occurrence of DNA aneuploidy, which was detected in 50% of patients, correlated significantly with tumor size and the presence of vascular invasion or intrahepatic metastasis. Overall, patients with DNA aneuploid tumors had a significantly worse prognosis than those with DNA diploid tumors (P less than 0.001) and, also in subdivided groups by tumor size (P less than 0.01). Among DNA aneuploid patients, the survival times were significantly shorter for patients with a low DNA index (less than 1.5) than for those with a high DNA index (greater than or equal to 1.5) (P less than 0.05). In a Cox multivariate analysis, nuclear DNA content provided significant prognostic value (P = 0.008), as did vascular invasion (P = 0.001) and intrahepatic metastasis (P = 0.005). These results indicated that nuclear DNA content has an important prognostic value in hepatocellular carcinoma.     

Acinic cell adenocarcinoma. A clinicopathologic analysis of 294 cases

Two hundred and ninety-four cases of acinic cell adenocarcinoma were reviewed for the purpose of defining the clinical parameters and determining the distribution of the four histomorphologic tissue patterns and five cell types for correlation to biologic behavior. The vast majority occurred in the parotid gland. There was a male predominance and a peak incidence in the third decade of life. The tumors were usually less than 3 cm in diameter and were slow growing. Pain was a common symptom, but was not indicative of prognosis. Nearly one half of the neoplasms exhibited multiple tissue growth patterns, and three fourths of the tumors displayed more than one cell type. The microcystic pattern was seen most frequently, regardless of the biological behavior of the tumors. The well-differentiated acinic cell was the most prevalent cell type except in cases with metastases, where the intercalated-duct cell type was slightly more frequent. Follow-up of 244 cases revealed a recurrence rate of 12%, a metastatic rate of 7.8%, and death rate of 6.1%. Since all histomorphologic patterns and cell types were manifest in tumors which recurred, metastasized, or caused the death of the patients, it seems appropriate to consider these neoplasms as low-grade adenocarcinomas rather than essentially benign with occasional unpredictable malignant behavior.     

Basaloid squamous carcinoma of the upper aerodigestive tract. Clinicopathologic and DNA flow cytometric analysis

This report adds nine basaloid squamous carcinomas (BSC) of the upper aerodigestive tract to the 11 already recorded in the literature. It includes the first flow cytometric analysis of their DNA content and compares the clinical behavior of BSC with conventional squamous cell carcinoma (SCC). An uncommon variant of squamous carcinoma, BSC manifests a predilection for the hypopharynx and base of tongue of men in the sixth decade of life. Histologically, the carcinoma is characterized by a basaloid pattern often in an intimate association with focal squamous differentiation, comedonecrosis, and stromal hyalinization. It is an aggressive neoplasm: seven of the nine patients had metastases to cervical lymph nodes at time of initial surgery and three of the five deaths occurred within 24 months after primary surgery followed by radiotherapy. Its aggressiveness notwithstanding, the biologic course of BSC is similar to that of conventional SCC when clinical stage, site, and treatment are matched. Patients with aneuploid BSC had a better mean survival time (39.5 months) than those with diploid carcinomas (16.3 months). Surgery followed by radiotherapy appears to be the treatment of choice. Because of a high incidence of distant metastases, adjuvant chemotherapy may be warranted.     

Renal cell carcinoma in children: a clinicopathologic study.

PURPOSE: To identify the prognostic factors, treatment, and outcome of children affected by renal cell carcinoma (RCC). PATIENTS AND METHODS: The series included 41 patients (18 males and 23 females) with a median age of 124 months observed at the 11 Italian Association for Pediatric Hematology and Oncology centers from January 1973 to January 2001. Clinical data, surgical notes, pathologic findings, and summaries of therapy were taken from the charts. RESULTS: Seven (17%) of the 41 patients had a papillary histology, and 34 (82.4%) had nonpapillary histology. Eighteen patients (43.9%) had stage I, one patient (2.4%) had stage II, two patients (4.8%) had stage IIIA, 10 patients (24.3%) had stage IIIB, and nine patients (21.9%) had stage IV disease. One patient had a bilateral involvement at diagnosis. Seven patients experienced disease recurrence. Lung and liver were the most common distant lesions and usually were fatal. In this study, the major factor influencing the prognosis was the stage. Event-free survival at 20 years was 53.5% for all patients. Overall survival at 20 years was 54.9% for all patients. CONCLUSION: RCC is a rare disease in children and adolescents. This neoplasm has a different clinical presentation in children compared with adults but the same outcome. In our experience, patients with localized disease could be cured by nephrectomy alone. Prospective studies in a larger number of patients are needed to confirm radiation therapy and biologic response modifiers as effective adjunct therapy in RCC stage III. The alternative therapy seems warranted in patients with advanced disease.     

Angiomatoid malignant fibrous histiocytoma: flow cytometric DNA analysis of six cases

Using flow cytometry, we examined the DNA content of six angiomatoid "malignant" fibrous histiocytomas (AMFH), seven nonangiomatoid MFH, and seven myxoid variants of MFH. All six AMFHs were diploid. All nonangiomatoid MFHs and six of seven of the myxoid variant were aneuploid. The clinical course, histological appearance, and ploidy pattern of AMFH are distinctly different from other MFH variants. These differences strongly suggest that AMFH is at the benign end of the clinicobiologic spectrum manifested by "malignant" fibrous histiocytomas.     

Deoxyribonucleic acid (DNA) ploidy and proliferative characteristics of metastatic squamous cell carcinoma determined by flow cytometric analysis.

BACKGROUND: The deoxyribonucleic acid (DNA) index and the proliferative index (the fraction of cells in the S phase) can be independent prognostic indicators of the biologic aggressiveness of certain malignant neoplasms. OBJECTIVE: To determine whether the DNA index or proliferative index could predict metastases in cutaneous squamous cell carcinoma. METHODS: Nineteen different metastases from 15 patients with primary cutaneous squamous cell carcinoma (SCC) were reviewed, graded, and had DNA proliferative indexing performed by fluorescent activated cytometry. A control group of 13 patients with primary cutaneous SCC without metastases were studied in a similar manner. RESULTS: No significant difference between the metastatic and the nonmetastatic SCC groups for either DNA index or proliferative index (non-paired t-test) was observed. No correlative association with histologic grading with DNA index or proliferative index was observed for either group. CONCLUSION: We conclude that aneuploidy and S-phase fraction by fluorescent activated cytometry are not significant predictors of potential metastases in cutaneous squamous cell carcinoma.     

Sarcomatoid renal cell carcinoma: clinicopathologic. A study of 42 cases

Forty-two cases of sarcomatoid renal cell carcinoma were reviewed clinicopathologically. Twenty-four patients were men, and 18 women; average age was 56.2 years (range, 30-81 years). Eight, 9, 13, and 12 cases were Stages I, II, III, and IV, respectively. Three morphologic patterns of sarcomatoid components were identified: malignant fibrous histiocytomatous (26 cases), fibrosarcomatous (6 cases), and unclassified sarcomatoid (10 cases). Mitotic count, degree of pleomorphism, cellularity, and amount of tumor matrix in the sarcomatoid areas, and similar morphologic parameters in the carcinomatous component all failed to correlate with prognosis, as did tumor size and renal vein involvement by tumor. Clinicopathologic stage was a most significant prognostic factor, with a survival of 49.7 months for Stage I and 6.8 months for combined Stages II, III, and IV. Tumor necrosis in the sarcomatoid area and proportion of sarcomatoid components were also poor prognostic factors. When these factors were compared to the stage, necrosis was an independent variable, however, proportion of sarcomatoid components was a poor prognostic indicator only for Stages I and II.     

Flow cytometric DNA analysis of squamous cell carcinoma of the esophagus

Although an abnormal amount of cellular DNA content is known to occur in squamous cell carcinoma of the esophagus, the significance and clinical relevance of this finding have yet to be elucidated. Paraffin-embedded blocks from 77 patients with squamous cell carcinoma of the esophagus were dewaxed and made into a suspension by mechanical mincing and enzymatic digestion. The nuclei were stained with propidium iodide, and the DNA content evaluated with flow cytometric study. The DNA index (DI) and the coefficient of variation were determined and the results were compared with other pathologic prognostic features. DNA aneuploidy was identified in 70.1% of the specimens and found to correlate significantly with both histologic grading (P less than 0.025) and the incidence of postoperative recurrence (P less than 0.05). Although the 5-year survival was slightly better in the euploid group than in the aneuploid group (28.8% and 19.1%, respectively), this difference was not statistically significant (P greater than 0.1). There was no correlation of the DI with the extent of wall penetration by the tumor, the incidence of lymph node metastases, or the surgical staging, features previously shown to correlate with prognosis in patients with esophageal carcinoma.     

Adenosquamous carcinoma of the stomach. A clinicopathologic analysis of 28 cases

Twenty-eight cases of primary adenosquamous carcinoma (ASC) of the stomach were studied clinicopathologically. These cases were classified into two types, 16 with differentiated type adenocarcinomatous component (DAC) and 12 with undifferentiated type adenocarcinomatous component (UAC), according to the degree of glandular formation of the adenocarcinomatous elements. A large number with adenocarcinomas, including 131 with differentiated type and 133 with undifferentiated type, were studied as controls. As a consequence, with respect to biologic behavior, ASC with DAC was similar to the differentiated type adenocarcinoma, and ASC with UAC to the undifferentiated type adenocarcinoma. Accordingly, the behavioristic feature of ASC seemed to be governed by the adenocarcinomatous component. Histologically, a close relationship between neoplastic adenomatous and squamous components was evident in the intermingling areas, thereby suggesting a transition of both elements. In addition, a mucoepidermoid pattern was occasionally detected in the squamous component. Judging from the biologic behavior and histologic findings, the majority of ASC probably derives from the squamous metaplasia in an adenocarcinoma. The prognosis of ASC was less favorable than that of adenocarcinoma because of the more extensive tumor depth and higher frequencies of lymphatic and vascular permeations of the carcinoma cells.     

Flow cytometric DNA analysis of interleukin-2 responsive renal cell carcinoma

Adoptive immunotherapy using interleukin-2 (IL-2) based therapy can result in marked tumor regression in some patients with metastatic renal cell carcinoma. DNA flow cytometry has not been previously studied as a predictor of outcome of this therapy. Archival paraffin embedded tumors were studied in 23 IL-2 treated patients with metastatic renal cell carcinoma. Eleven patients were complete responders (CR) and 12 were nonresponders (NR). In the CR group, 4/11 (40%) were diploid and 7/11 (60%) were aneuploid. In the NR group, 9/12 (75%) were diploid and 3/12 (25%) were aneuploid. Although there was a trend that patients with an aneuploid DNA pattern were more likely to undergo a complete response, ploidy pattern alone was not significantly predictive of response (p² = 0.10, Fischer's exact test). When combining ploidy pattern with other variables that were predictive for complete response, such as good performance status and a higher pretreatment weight, prediction of complete response was not improved by including ploidy. This preliminary report suggests that DNA ploidy does not appear to provide any additional information concerning responsiveness to IL-2 based immunotherapy beyond that obtained by performance status and pretreatment weight in this patient population. © 1993 Wiley-Liss, Inc.     

Synovial sarcoma. A DNA flow cytometric study

The relationship between DNA content, clinicopathologic findings, and patient survival in synovial sarcoma was investigated. Patient age at diagnosis (P less than 0.001), tumor size (P less than 0.001), and ploidy status (P less than 0.003) correlated significantly with patient survival. A marginally significant correlation between mitotic count and patient survival was also observed (P = 0.04). Histologic subtypes (monophasic versus biphasic), mitotic count, and S-phase by flow cytometry had no significant influence on the clinical outcome of patients with synovial sarcoma in this study. The authors conclude that DNA ploidy analysis is a significant objective probe in the prognostication of patients with synovial sarcoma.