Cytophotometric analysis of the nuclear DNA content in oral squamous cell carcinoma--relation between nuclear DNA content and clinico-pathologic findings

Cytophotometric analysis has been performed on Feulgen-stained nuclei in smears taken from 48 biopsy specimens of squamous cell carcinoma of the oral cavity. The results were correlated with the clinico-pathologic findings of the tumor. DNA histograms were graded into three patterns: P-I (2C-2C), P-II (2C-6C) and P-III (2C-beyond 6C). DNA patterns of the carcinoma of the clinical stage I, II tended to be P-I, II and those of stage III, IV were mostly P-II, III. DNA patterns of carcinomas which were well differentiated and poorly pleomorphic with a few number of mitoses, were mostly P-I, whereas those which were poorly differentiated and remarkably pleomorphic with a many number of mitoses were mostly P-III.     

Prognostic significance of nuclear DNA content in human neuroepithelial tumors

The relationship between survival and flow cytometric DNA-ploidy and other prognostic factors such as histological subtype, anatomical tumor site, patient sex and age was investigated in 153 patients with intracranial neuroepithelial tumors who underwent surgical treatment. We found a trend toward poorer survival from anaplastic astrocytomas and glioblastomas with respect to low-grade (I and II) astrocytomas (which did not differ significantly); accordingly, patients were grouped into these 3 histologic subgroups. Thirty-seven of the 153 tumors (24.2%) were aneuploid with a median DNA-index (DI) of 1.3 (range: 1.2-2.0). DNA-ploidy correlated with histology, since anaplastic astrocytomas and glioblastomas were significantly (p = 0.041) more frequently aneuploid (around 30%) than low-grade astrocytomas (around 10%). Patients with DNA-aneuploid tumors (i.e., with DI not equal to 1.00) survived for a shorter time (31.4 weeks) than patients with DNA diploid tumors (75.1 weeks) (p less than 0.001). This difference was confirmed by Cox's multivariate analysis. Aneuploid tumors were associated with a poorer survival (p = .0002) when compared with diploid tumors, resulting in a relative risk point estimate (RR) of 2.41, 95% confidence interval (Cl) = 1.55-3.74. Histological subtype was also significantly associated with survival (p less than 0.0001), with RRs of 2.09, 95% Cl = 1.13-3.86 and 3.59, 95% Cl = 1.96-6.59 for anaplastic astrocytomas and glioblastomas, respectively, compared to low-grade astrocytomas. We therefore suggest that the flow cytometric measurement of DNA-ploidy has relevant significance in predicting survival in patients treated for intracranial neuroepithelial tumors.     

Cytophotometric DNA content and argyrophilic nucleolar organiser regions of oesophageal carcinoma.

The cytophotometric DNA content and the argyrophilic nucleolar organiser regions (AgNORs) of biopsy specimens taken before undergoing any treatment were examined in 91 surgically treated oesophageal carcinoma cases. There was a significant linear dependence between the mean DNA content and the number of AgNOR per nucleus (AgNOR number) (r = 0.615, P < 0.001). The DNA distribution pattern and the range of the AgNOR number also showed a significant correlation (P < 0.01). Twenty three of 28 cases with a low AgNOR number (< 4) were then determined to have a diploid pattern (type II), while 17 out of 22 cases with a high AgNOR number (> or = 6) had high ploidy values (type IV). The patients with a type II DNA distribution pattern and a low AgNOR number thus showed a good post-operative course with a 5 year survival rate of 55.2%, whereas no patients survived over 4 years among the 17 cases with both a type IV DNA pattern and a high AgNOR number (P < 0.001). These data thus demonstrate the close relationship between cytophotometric DNA content and AgNOR number and suggest that the combined detection of these two parameters, using biopsy specimens, should be of benefit in making an accurate preoperative evaluation of prognosis for patients with oesophageal carcinoma.     

The prognostic significance of nuclear DNA content in malignant epithelial tumors of the ovary

Recent studies have indicated that the nuclear DNA content of certain malignant neoplasms can be used as an adjunct in predicting their biologic behavior. The DNA content of 99 ovarian carcinomas was determined by flow cytometric analysis of nuclei obtained from paraffin-embedded tissue. Of the 99 tumors, 51 were diploid and 48 showed one or more aneuploid peaks. The 5-year survival for patients with diploid tumors (50%) was significantly higher than for patients with aneuploid tumors (22%) (P less than 0.01). Other factors which significantly affected survival were clinical stage (P less than 0.001), tumor pattern grade (P less than 0.01), DNA index (P less than 0.01), the presence of ascites (P less than 0.001), peritoneal carcinomatosis (P less than 0.0001), and residual tumor at second-look laparotomy (P less than 0.05). Diameter of the primary ovarian tumor, diameter of the largest peritoneal implant before debulking, and the percent S-phase had no significant correlation with survival. Of 16 patients with aneuploid tumors who underwent second-look laparotomy, nine (56%) had residual tumor, compared to six of 22 of patients with diploid tumors (27%). Of seven patients with aneuploid tumors and a negative second-look laparotomy, four (57%) died from recurrent tumor. By comparison, of 16 patients with diploid tumors and a negative second-look laparotomy, only four (25%) died from recurrent tumor. The determination of DNA ploidy in ovarian carcinomas may be used as an adjunct in predicting tumor behavior, response to chemotherapy, and late recurrence of disease.     

Cytologic study of glandular tumors in maxillofacial regions-- diagnostic application of nuclear DNA content

We studied whether the DNA contents may be useful in the differential diagnosis of glandular tumors in the maxillofacial regions. In 25 of these tumors, the DNA contents were measured by microspectrophotometry, using 4 normal salivary glands as controls. We found that: (1) DNA histgram patterns were useful in differential diagnosis only in special cases. (2) The criteria were based on our decision and these tumors were classified into 3 types (a) Benign, (b) Low-grade malignant, (c) High-grade malignant. Our classification seemed to offer an objective means for differentiating between Benign & Malignant tumors of these types.     

Photometric studies on nuclear DNA content and area in different laryngeal epithelia

A photometric study of different laryngeal epithelia was performed. The nuclear DNA content and area for epithelia with keratosis, hyperplasia, and moderate dysplasia were comparable to normal. These lesions cannot therefore be considered to be premalignant photometrically. The photometric values for carcinoma in situ and severe dysplasia were comparable to that for invasive carcinoma. Long-term follow-up has to be performed to evaluate the biological significance of these different lesions.     

Flow cytometric analysis of the nuclear DNA content of hepatoblastoma.

The nuclear DNA content of 15 hepatoblastoma cases was determined in paraffin-embedded tissues by flow cytometry. The DNA index (DI) was calculated, and the ploidy pattern of nuclear DNA was estimated. The correlation between the ploidy pattern and clinicopathologic findings was studied, and the prognostic significance of the ploidy pattern was investigated. An aneuploid pattern was seen in 50% of the lesions with histologic embryonal and anaplastic types. It was not seen in the fetal type. In the tumors with combined epithelial components, the fetal-type component had a diploid pattern in all five cases. The embryonal-type component was associated with aneuploidy in two of five cases. In aneuploid tumors, vascular invasion (tumor emboli in the vessels) was observed more frequently. The prognosis of the patients with an aneuploid tumor was significantly poorer. These results indicate that nuclear DNA ploidy pattern analysis might be useful in investigating the prognosis of hepatoblastoma.     

The prognostic significance of nuclear DNA content in prostatic carcinoma.

The prognostic significance of ploidy level was studied in prostatic carcinoma and compared with the prognostic significance of morphologic grade and clinical stage. A nonselected group of 145 patients was studied in whom prostatic carcinoma was diagnosed by fine-needle aspiration biopsy at the Karolinska Hospital in 1966. All patients had endocrine therapy and were observed for 23 years or until death. Ploidy level was determined from cytophotometric measurements of Feulgen-stained tumor cell nuclei. The original May-Grünwald-Giemsa-stained cytologic slides, on which the cancer diagnoses were based in 1966, were destained and subsequently Feulgen stained for cytophotometric analysis. From the Feulgen-DNA data, the tumors were classified as near-diploid, near-tetraploid, and highly aneuploid, variants D-type, T-type, and A-type, respectively. The A-type tumors progressed rapidly, and 96% of the patients with this type died of the tumor within 5 years and all patients with this type died within 7 years. D-type and T-type tumors progressed much more slowly. None of the patients with these types of tumors died of the tumor disease within the first 5 years after diagnosis. As many as 12% of the patients (crude survival rate; corresponding to a relative survival rate of 43%) were still alive 15 years after diagnosis. According to multivariate Cox regression analysis, ploidy compared with grade and tumor stage was the strongest predictor of survival.     

Benign cartilaginous tumors of the upper airway

Benign cartilaginous neoplasms of the laryngotracheal apparatus are uncommon clinical entities. Two cases of cartilaginous lesions of the upper airway are reported. Resection with maintenance of upper airway structural integrity is the preferred treatment. Temporary tracheostomy is often necessary and can provide access for stenting of the tracheal repair.     

Nuclear DNA content and nuclear atypia. Relation to survival in patients with breast adenocarcinoma and serious ovarian tumors

The grade of nuclear atypia (low, medium, or high) was compared with DNA content in 122 tumors comprising 73 serous ovarian tumors and 49 mammary carcinomas of ductal type. The study was performed on two consecutive histologic sections 4 mu in thickness, one of which was stained with hematoxylin-eosin for estimation of nuclear atypia, and the other with Feulgen for measurements of DNA. In both tumor groups there was a significant correlation between nuclear atypia and nuclear DNA content. The results suggest that DNA analysis is of significant additional prognostic value even when the grade of nuclear atypia is known.     

Flow cytometric analysis of pituitary tumors. Correlation of nuclear antigen p105 and DNA content with clinical behavior

Flow cytometric quantitation of the proliferation-associated nuclear antigen p105 and DNA content was performed on nuclear suspensions from 12 paraffin-embedded pituitary macroadenomas and one pituitary carcinoma and correlated with clinical outcome. Median follow-up was 41 months (range, 33 to 48 months). Three of the 13 tumors (23%) had an identifiable aneuploid peak. Of the four tumors that recurred or metastasized, only one was aneuploid. Nuclear antigen analysis of all diploid tumors showed enhanced p105 expression in G2M phase cells compared to G0G1 cells. The G2M/G0G1 fluorescence ratio for p105 was consistently higher (P less than 0.05) for the three diploid tumors that recurred (median, 1.32 arbitrary fluorescence units; range, 1.27 to 1.80) than for the seven nonrecurrent diploid tumors (median, 1.20 arbitrary fluorescent units; range 1.14 to 1.22). These findings indicate a low incidence of DNA aneuploidy among pituitary tumors and suggest that for diploid adenomas, measurement of p105 may provide information useful in predicting prognosis and directing postoperative adjuvant therapy.     

Cytophotometric DNA measurements in medullary thyroid carcinoma

The prognostic significance of objectively measured karyometric variables (ploidy pattern, nuclear roundness, area, elongation, chromatin texture, and nearest nucleus distance) was investigated in relation to clinical (stage and type of disease) and morphologic (histologic patterns) variables in 27 patients with the diagnosis of medullary thyroid carcinoma (MTC). The DNA and karyometric measurements of Feulgen-stained nuclei were made with a video cytometry system. The five-year and ten-year adjusted survival rates were 74.4 +/- 10.1% and 59.5 +/- 15.6%, respectively. Cox's survival analysis for mortality showed that only stage, age, sex, and 5N exceeding rate had predictive value (overall P = 0.0012) in decreasing order. Patients with the best prognosis were young females with clinical Stage I disease and low 5N exceeding rate tumors. When karyometric and histometric variables were considered by themselves survival correlates with the standard deviation (SD) of the nearest nuclear distance and nuclear elongation; that is, patients with crowded, high cellularity tumors and elongated cells had the worst prognosis. In univariate analyses only clinical stage correlated with adjusted survival rate. Multivariate survival analysis for morbidity showed that patients in Stages greater than or equal to II and high SD of ploidy values were free of symptoms for short intervals. When morphometric data were considered alone, patients with high variance in the chromatin texture and highly variable nuclear areas had shorter asymptomatic intervals.     

Prognostic significance of nuclear DNA content in breast cancer

The relationship between nuclear DNA content determined by cytofluorometry in the primary focus of breast cancer and the survival rate was analyzed to clarify the prognostic significance of nuclear DNA content in breast cancer. The relationships of the ploidy pattern and the frequency of polyploid cells (4c or above) with the survival rate were studied in patients who underwent extended radical mastectomy and were comparable in the clinical stage and other prognostic factors. The survival rate was significantly lower in those of the non-diploid type showing no prominent peak at 2c or those in whom the frequency of polyploid cells was more than 30% than in those of the diploid type with a prominent peak at 2c or those showing few polyploid cells, even when the disease stage (Stage II by TNM classification and stage III by Tnm classification), histological lymph node metastasis (n (+), n1 beta), and histological type (papillotubular carcinoma, scirrhous carcinoma) were identical. From these findings, nuclear DNA content is considered to be a parameter of the malignancy of breast cancer and to have clinical significance as an important prognostic factor.     

Cytophotometric studies on mycosis fungoides and other cutaneous reticuloses.

Feulgen-DNA cytophotometry was carried out in skin imprint preparations of patients with mycosis fungoides, reticulum-cell sarcoma, lymphomatoid papulosis and Sézary's syndrome. These patients proved to have aneuploid and polyploid DNA histograms. In tumour stage of mycosis fungoides a more bimodal distribution of the DNA values was found. Of 79 cases of suspected malignant reticulosis of the skin, 32 showed an abnormal DNA histogram with 5 percent or more tetraploid and hypertetraploid cells. Thirty of these patients developed a malignant skin reticulosis during the 4-year follow-up period. It is concluded that DNA cytophotometry has value as a supplement to routine morphological histopathology in malignant cutaneous reticulosis.