DNA Aneuploid Pattern Research Articles

Surgery for brain metastases from nonsmall cell lung carcinomas and tissue cultures from the resected specimens

Between 1978 and 1989, 44 patients underwent 44 thoracotomies and 55 craniotomies for nonsmall cell lung carcinoma (NSCLC) and its brain metastases. Patients ages ranged from 20 to 75 years. There were no intraoperative mortalities. The 2-, 3-, and 5-year survival rates following the initial craniotomy were 23%, 10%, and 10%, respectively. Patient survival did not differ with respect to solitary or multiple metastases or the sequence of surgery for primary lesion and brain metastases. Moreover, there was no significant difference in survival between patients treated by surgery alone and those receiving surgery followed by whole brain radiotherapy. After 1985, in vitro tissue culture was attempted using freshly resected specimens of brain metastases obtained from 30 consecutive cases. Of those specimens, nine (30%) were successfully established as permanent cell lines. Eight of those cell lines revealed DNA-aneuploid pattern on flow cytometric analysis. The remaining cell line was not analyzed. Karyotype analysis was also performed in eight of nine established cell lines. Two adenocarcinoma cell lines showed the presence of +3p- chromosome, and three showed +7q- chromosome as recurrent chromosomal abnormalities. These findings provide new evidence concerning the presence of 3p- and/or 7q- marker chromosomes in certain adenocarcinoma cell lines established from brain metastases. The prognosis was poorer in the group with in vitro tumor growth than that in the group showing no in vitro tumor growth. These cell lines established from brain metastases may be useful materials not only for studying the biological characteristics and chemo-sensitivity testing, but also for estimating prognoses after resection of brain metastases.

Flow Cytometric DNA Analysis of Submucosal Carcinoma of the Esophagus

Carcinoma of the esophagus confined to the submucosa presents clinicopathologic features different from those of either carcinoma limited to the mucosa or more advanced lesions. The present study was designed to find out whether or not analysis of DNA content would yield prognostic information allowing the clinical outcome of carcinoma of the esophagus confined to the submucosa to be predicted. Cell nuclear DNA content was determined by flow cytometry in paraffin-embedded blocks from 55 patients with carcinoma of the esophagus confined to the submucosa. This allowed classification of DNA content into diploid and aneuploid patterns based on combined DNA histogram and cytogram data. The aneuploid pattern included typical, multiple clone and small clone aneuploid subtypes, as classified by our new criteria. A DNA aneuploid pattern was identified in 26 cases (47.3%) and found to correlate significantly with lymph node metastasis (P < 0.01). The incidence of distant metastasis was significantly higher in the aneuploid (4/26) than in the diploid (0/29) group (P < 0.05). The mortality from cancer within two years of surgery was significantly higher in the aneuploid (4/26) than in the diploid (0/29) group (P < 0.05). In conclusion, flow-cytometric DNA analysis is a valuable method for predicting malignant potential in carcinoma of the esophagus confined to the submucosa.

Prognostic importance of DNA flow cytometric variables in rhabdomyosarcomas.

To determine whether DNA ploidy patterns and S phase fraction offer prognostic information in patients with rhabdomyosarcoma (RMS). DNA flow cytometry was performed on formalin fixed, paraffin wax embedded samples from primary tumours, and metastatic deposits or recurrences in 70 patients. DNA histogram analysis was done using a semi-automated cell cycle analysis program. Of the 70 primary tumours, 23 were DNA diploid, 32 DNA aneuploid, eight DNA multiploid, and seven DNA tetraploid. The prognosis for DNA aneuploid patterns was favourable, intermediate within the group of DNA tetraploid tumours and poor among patients with DNA diploid and DNA multiploid tumours (p = 0.009). In multivariate analysis (Cox regression model) DNA ploidy was an important independent prognostic factor, along with TNM stage, localisation, and histopathological classification. Ten out of 32 patients with a high S phase fraction (> 15%) with primary RMS achieved long term survival in contrast to 20 out of 29 patients with a low S phase fraction (< or = 15%) (p = 0.008). In 24 cases the DNA ploidy of cases of relapse was analysed. Of the 15 cases, in which stem line changes had occurred, 13 died of disease. No stem line changes were noted in nine cases and in this group four patients died of disease (p = 0.02). Assessment of DNA ploidy and S phase fraction in primary RMS and evaluation of stem line changes in cases of relapse are important variables in predicting prognosis.

Flowcytometric DNA ploidy pattern for predicting metastasis of clinical stage I nonseminomatous germ cell testicular tumors

The best treatment of patients with clinical Stage I nonseminomatous germ cell testicular tumors (NSGCTT) remains controversial. Archival formalin fixed paraffin-embedded testicular tumor specimens from 23 patients diagnosed between 1977 and 1988 were available for analysis. All patients had clinical Stage I NSGCTT, and the specimens were prepared using Hedley's technique and propidium iodide staining prior to flow cytometric ploidy analysis. Tumors from 3 patients (13 %) were classified as DNA diploid; the remaining 20 tumors had DNA aneuploid patterns. Eleven patients had retroperitoneal lymph node dissection (RPLND); 2 (18 %) were upstaged to pathologic NSGCTT Stage IIA, and 1 had a subsequent relapse in the contralateral testicle. Among 12 patients in the surveillance protocols, 3 (25 %) had tumor recurrence. All patients who were either upstaged to pathologic NSGCTT Stage IIA or had tumor recurrence while under surveillance had DNA aneuploid patterns. The 3 patients with DNA diploid tumor patterns had been in the surveillance protocols. To date, with fifteen months minimum follow-up, none has had tumor recurrence. Such data suggest that flow cytometric DNA ploidy analysis, when used as an adjuvant to more traditional prognostic parameters, may help to identify a group of patients with clinical Stage I NSGCTT at very low risk for recurrence and perhaps well-suited for surveillance management.

Oncocytic tumors of salivary gland type: a study with emphasis on nuclear DNA ploidy.

We studied nine cases of oncocytic tumors of salivary gland type in order to evaluate their clinico-pathologic profiles and nuclear DNA patterns as criteria for the differential diagnosis between benign and malignant forms. The tumors were located at the parotid gland, palate, and orbit. The age of the patients ranged between 35 and 85 years and the sex ratio (F:M) was 1:0.8. Seven tumors were capsulated and had typical cytology: they were composed of polyhedrical cells with large, eosinophilic, and granular cytoplasm and dark nucleus. The remaining two tumors exhibited malignancy criteria for the oncocytic tumors: atypia, pleomorphism, and mitosis. The evaluation of the nuclear DNA content was also distinct: benign tumors had a DNA diploid pattern and the malignant neoplasms displayed a DNA aneuploid pattern. These observations point to DNA nuclear assessment as an additional criterion to discriminate neoplasms with divergent clinical behavior and prognosis.

Flow cytometric DNA analysis of advanced prostatic adenocarcinoma.

Thirty-two patients with extracapsular invasive prostatic adenocarcinoma were analysed to investigate the prognostic significance of DNA flow cytometry by using the nuclear material extracted from paraffin embedded prostatectomy specimens. The DNA ploid pattern was diploid in 61% of the tumors and aneuploid in 39%. A significant correlation between the DNA diploid pattern with pathologic low grade tumors and the DNA aneuploid pattern with high stage tumors was not proved. The clinical stage correlated well with patients survival but only DNA ploidy pattern and pathologic grade had trends of correlation. However, if only the stage D tumors were analysed, a significant prognostic predictive value of DNA flow cytometry was identified. Although the results maybe regarded as preliminary, (in view of the small number of patients), these suggest that DNA content from the initial surgical samples provide only a synergistic effect to the clinical stage and pathologic grade in the prognostic evaluation of the advanced prostate cancers especially for those who underwent hormonal therapy postoperatively.

喉頭の前癌病変の解釈

Three parameters of epithelial dysplasia of the larynx, namely, the expression of oncogene product (ras p21, EGF-R), nuclear DNA contents measured by flowcytometry and the proliferating activity (Ki-67, PCNA), were examined to determine the appropriate clinical treatment and the prognosis. Dysplasia was classified into low grade and high grade depending upon the degrees of cellular atypia and polarity-disorder in the depths of the epithelium. Fourteen of 20 cases with high grade dysplasia and 7 of 21 cases with low grade dysplasia were positive to ras p21. Two of 6 cases with high grade dysplasia and 1 of 5 cases with low grade dysplasia were positive to EGF-R. Four of 8 cases with high grade dysplasia and 2 of 7 cases with low grade dysplasia had a DNA aneuploid pattern as determined by flow cytometric DNA analysis. High grade dysplasia had higher Ki-67 and PCNA labeling indeces than low grade dysplasia. These three parameters may be useful in deciding the therapeutic modalities and the prognosis of dysplasia.

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.

DNA ploidy as a predictive index of therapeutic response in lung cancer.

Background: Although many authors have reported that the median survival time of surgically resected non-small cell lung cancer (NSCLC) was shorter in aneuploid than in diploid determined by flow cytometry, there are few reports about DNA ploidy using bronchial brushing material in all types of lung cancer. Method: The DNA ploidy test results of 109 consecutive patients with lung cancer were analyzed to find the relationship of DNA ploidy and anatomic or physiologic stage. And the differences of the response to various therapeutic modalities according to DNA ploidy were evaluated at least 8 weeks after the begining of the therapy. Results: Numbers of patients with DNA aneuploid pattern or high proliferative activity (S+G2M>22%) were not different among the various cell types of lung cancer. The relationship of DNA ploidy and anatomic or physiologic stage was not significant. However, NSCLC patients with high proliferative activity showed more advanced anatomic stage than those without that (p

Study of cellular DNA content by flow cytometry in primary bladder carcinomas. Significance of monoclonal and multiclonal varieties of DNA aneuploidy.

A prospective study of cellular DNA content by flow cytometry was performed on a nonconsecutive series of 67 patients undergoing diagnostic and/or therapeutic transurethral resection for primary urothelial bladder carcinoma. DNA-aneuploidy was present in 82% of the cases (55/67), while multiclonality was found in 45% of the DNA-aneuploid cases (25/55). DNA-ploidy was much more strictly correlated with histological grading (p less than 0.005) than with papillary or non-papillary growth pattern (p less than 0.05) or T staging (p less than 0.05). Of 26 patients with a minimum follow-up of 24 months, 100% (6/6) of cases with DNA-diploid neoplasias showed no signs of disease relapse, versus 10% (2/20) of those with DNA-aneuploid neoplasias (p less than 0.001). Furthermore, tumoral progression occurred in 10 of 20 cases (50%) with aneuploid DNA content. In this latter group of 10 cases a multiclonal DNA-aneuploid pattern was found, with a significant difference (p less than 0.001).

Flow cytometric analysis in colorectal carcinoma: prognostic significance of cellular DNA content

The prognostic value of DNA ploidy status was evaluated prospectively in 70 patients with colorectal carcinoma. Cellular DNA content was measured by flow cytometry from fresh specimens with multiple site sampling. Seventy-five percent of cases exhibited a DNA aneuploid pattern. In a univariate analysis, DNA ploidy status showed a statistically significant correlation with survival (p less than 0.05), weaker than Dukes' stage (p less than 0.001). No correlation was observed between survival and presence of multiple DNA stemlines. In a multivariate analysis, Dukes' stage was the strongest prognostic indicator (p = 0.01) while DNA ploidy status did not show an independent prognostic value. It is concluded that DNA ploidy status is associated with pathological features of aggressive malignancy, but it does not have a determinant role in predicting survival.

Prognostic and therapeutic significance of the flow cytometric nuclear DNA content in non-small cell lung cancer

To evaluate prognostic and therapeutic significance, tumor DNA content was determined by flow cytometry in 310 paraffin-embedded tissue samples obtained surgically from 130 patients with non-small cell lung cancer. Ninety-six (76.8%) patients had DNA aneuploid patterns that were statistically higher in adenocarcinoma than in squamous cell carcinoma. A better 5-year survival rate was observed in Group A (DNA diploidy, 69.6%) than in Group B (DNA aneuploidy and DNA peridiploidy, 33.2%; P less than 0.001). The survival curves of the patients in Group B continued to decrease during the next 2.5 years. Cox's model analysis showed that both the pathologic stage and the DNA content were the significant prognostic factors for survival. However, the DNA content was an independent prognostic factor in squamous cell carcinoma, but not in adenocarcinoma. These results indicate that DNA content analysis is useful for the evaluation of clinical behavior and prognosis, and that the clinical value of the DNA content must be differentiated between squamous cell carcinoma and adenocarcinoma.

Flow Cytometry による腎細胞癌の Heterogeneity の検討

DNA content in renal cell carcinoma was investigated to examine the tumor heterogeneity and was correlated with their morphologic grades. A total number of 147 paraffin-embedded samples (2-6 samples with a mean of 5) from 30 tumors were analyzed by flow cytometry. DNA aneuploid patterns were demonstrated in 5 of 9 grade 1 tumors (56%), 15 of 18 grade 2 tumors (83%) and 3 of all grade 3 tumors (100%), while aneuploid DNA histograms were exhibited in 20 of 57 grade 1 samples (35%), 45 of 81 grade 2 samples (56%) and 8 of 9 grade 3 samples (89%). DNA aneuploid patterns were demonstrated more significantly in grade 2 and 3 samples than in grade 1 samples (p less than 0.02 and p less than 0.01, respectively). Consequently, 73 samples (50%) showed DNA aneuploid patterns and 74 samples exhibited DNA diploid patterns. Eleven tumors (37%) showed homogeneous DNA ploidy patterns (7 tumors were diploid and 4 tumors were aneuploid only), while 19 tumors (63%) showed DNA heterogeneity, 17 of these 19 tumors demonstrating diploid as well as aneuploid samples. DNA heterogeneity was not found when the tumor samples examined were fewer than 3 samples. However, 19 of 25 tumors (76%) in which more than 4 samples were examined showed DNA heterogeneity, the incidence of which tended to be increased with upgrading of the tumor--3 of 9 (33%) in grade 1 tumors, 13 of 18 (72%) in grade 2 tumors and 3 of all (100%) in grade 3 tumors.(ABSTRACT TRUNCATED AT 250 WORDS)

Stage C Prostatic Adenocarcinoma: Flow Cytometric Nuclear DNA Ploidy Analysis

Flow cytometric nuclear DNA ploidy analysis was used to study pathologic stage C prostatic adenocarcinoma (pT3, N0, M0) in 146 patients who underwent radical retropubic prostatectomy and bilateral pelvic lymphadenectomy between 1967 and 1981. Of these tumors, 46% had a DNA diploid pattern, 47% had a DNA tetraploid pattern, and 7% had a DNA aneuploid pattern. Abnormal ploidy patterns were associated more frequently with histologic high-grade tumors than with low-grade tumors. Considered alone, DNA ploidy pattern showed a strong association with subsequent prognosis. The median interval to progression for tumors with DNA tetraploid and DNA aneuploid patterns was 7.8 and 3.5 years, respectively. For the DNA diploid tumors, only 23% progressed within 18 years, the longest follow-up. At 10 years, only 10% of patients with DNA diploid tumors had died of prostatic cancer, in comparison with 28% of the DNA tetraploid and 36% of the DNA aneuploid groups (P less than 0.01). By analysis of a combination of histologic tumor grade and nuclear DNA ploidy pattern, an even stronger association with prognosis was demonstrated. For the 38 patients with histologic low-grade and DNA diploid tumors, progression-free survival was 92% at 10 years, in comparison with 57% for 23 patients with low-grade DNA nondiploid tumors. Patients with high-grade tumor had a poorer prognosis whether the DNA ploidy pattern was diploid or nondiploid. Nuclear DNA ploidy pattern is an important and independent prognostic variable for patients with pathologic stage C prostatic cancer treated by radical prostatectomy.