Value Of DNA Flow Cytometry Research Articles

Hypoploidy defines patients with poor prognosis in breast cancer

The clinicopathological features currently used in breast cancer prognosis often fail to characterize the clinical heterogeneity of the disease accurately. Our study is aimed to investigate the predictive value of DNA flow cytometry in breast cancer. Previously untreated breast carcinoma samples (584) were snap frozen for flow-cytometry. Tumors were classified into three DNA index (DI) categories: i) tumors showing a DI =0.96-1.15 (diploid and near-diploid); ii) tumors with a DI >or=1.16 (hyperdiploid, tetraploid, multiploid and/or those with more than one diploid population); and iii) tumors with a DI <or=0.95 (hypoploid). The 5- and 10-year cumulative survival rates +/- SE for Group I (n=191) were 98+/-1% and 98+/-1%. For Group II (n=361) these rates were 77+/-2% at 5 years and 63+/-5% at 10 years. In Group III (n=32) the rate at 5 years was 23+/-8%, with no patients alive at 10 years (p<0.0001). In univariate analysis, tumor size, node status, grade, karyometry, S-phase fraction, MIB-1 index, and estrogen receptors retained prognostic significance; in multivariate analysis, only DI <or=0.95 (hypoploid) was retained as an independent prognostic factor for overall survival. Our data strongly support that DNA hypoploid has a strong, independent prognostic value for predicting the short-term clinical outcome of breast carcinoma patients.

Flow cytometric DNA ploidy in salivary gland tumours

This study on 279 tumours of the salivary glands was conducted to analyse whether the assessment of DNA ploidy by flow cytometry may assist histopathology in discriminating benign from malignant types of tumours. The group of benign tumours included 164 pleomorphic adenomas, 51 Warthin's tumours, 7 basal cell adenomas, 2 lipomas as well as 5 other different tumours. All of the 229 benign tumours were diploid. The malignant tumours consisted of 18 adenoid cystic adenomas, 10 mucoepidermoid carcinomas, 5 acinic cell carcinomas, 5 carcinoma in pleomorphic adenoma as well as of 12 other malignancies belonging to 7 different tumour entities. Twelve of 50 malignant salivary gland tumours were aneuploid. There was no significant relationship between the DNA ploidy status and histopathological grading, lymph node metastasis and local recurrence development, respectively. In three cases which initially were taken for pleomorphic adenomas by routine histological examination, aneuploid cell populations exposed by DNA flow cytometric analysis gave rise to a closer inspection of the suspect lesions. Examination of consecutive slides actually revealed small assemblies of carcinoma cells that required a final diagnosis as non-invasive carcinoma in pleomorphic adenoma. The most obvious value of DNA flow cytometry in salivary gland tumours is thus its contribution to assist histopathology in identifying potentially malignant lesions.

DNA flow cytrometry in solid tumors

This brief overview outlines the fundamental principles of flow cytometry with emphasis on DNA measurements and cell cycle analysis in human solid tumors. Type of material used, sampling processing procedures and methods of analysis of data are discussed. DNA ploidy and proliferative activity (S-phase fraction) are the two biological parameters commonly measured by DNA flow cytometric analysis. The prime purpose of most studies in this area is the investigation of the prognostic value of DNA flow cytometry in addition to the information provided by conventional clinicopathological factors known to affect disease prognosis. Numerous studies concerning the predictive significance of DNA flow cytometry in some types of solid tumors are reviewed in this article. The general statement, for tumors in the same histopathological stage of the disease, is that diploid and/or low proliferative tumors have a more favourable prognosis than aneuploid and/or high proliferative tumors, suggesting an important role of DNA flow cytometry in the assessment of tumor behaviour and in the outcome evaluation of the disease. However, in some studies this association could not be substantiated, and the prognostic relevance of DNA analysis has been questioned. The potential reasons for conflicting results, namely the methodological pitfalls related to the cell preparation techniques and the histogram interpretation are discussed.

A prospective correlation of Laurén's histological classification of stomach cancer with clinicopathological findings including DNA flow cytometry.

Between November 1990 and December 1992, 217 patients with stomach cancer were enrolled in a prospective study evaluating the prognostic value of DNA flow cytometry. Laurén's histological type was evaluated in 216 cases, of which 102 (47%) were of the diffuse type, 74 (34%) were of the intestinal type, and 40 (19%) were mixed type tumors. Laurén's histological type showed a significant correlation with age (p = 0.028), sex (p = 0.004), tumor size (p = 0.002), T stage (p = 0.006), overall TNM stage (p = 0.008), histological grade (p < 0.001), and tumor ploidy (p < 0.001). Intestinal type stomach cancer showed a significantly higher proportion of aneuploidy [diffuse vs. intestinal type; 41/102 (40%) vs. 52/74 (70%)]. After a median follow-up of 66.1 months (range, 29.6-78.1), 110 of 216 patients (51%) survived. Patients with intestinal type stomach cancer had a significantly better survival than did those with diffuse type stomach cancer (64% vs. 42% of patients surviving, p = 0.020). Our study suggests that there are biological differences between the two subtypes of Laurén's classification of stomach cancer in addition to the morphological differences. Laurén's classification should remain valid in future studies investigating the pathogenetic and clinical aspects of stomach cancer.

Role of DNA flow cytometry and image cytometry on effusion fluid

The objective of the study was to assess the value of DNA flow cytometry (FCM) and image cytometry (ICM) as an adjunct to routine diagnostic cytology. In this prospective study, 100 consecutive effusion fluids were studied for routine cytology, DNA FCM, and in selected cases, ICM. One half of the centrifuged fluid sample was used for routine cytology and the remaining portion was used for DNA FCM. Nuclear area, nuclear diameter, nuclear perimeter, nuclear convex perimeter, nuclear roundess, and nuclear convex area were measured on at least 100 cells by ICM in cytologically malignant or DNA aneuploid cases along with control cases. Clinical follow-up was done in all cases. There were 22 cytologically malignant cases and 78 cytologically benign cases. Among the 22 cytologically malignant cases, there were 11 aneuploid and diploid cases each by DNA FCM. Out of 78 cytologically benign cases, six (7.7%) were aneuploid by DNA FCM. Smears of these cases showed predominantly reactive mesothelial cells, but the DNA histograms showed hypodiploid (one), hyperdiploid (three), tetraploid (one), and hypertetraploid (one) aneuploidy. Follow-up of these cases showed clinical or histologic features of malignancy except in one case of tetraploid aneuploidy, which did not show any features of malignancy and responded well to antitubercular therapy. Therefore, out of 27 malignant effusions, DNA FCM picked up 16 cases and routine cytology detected 22 cases. Sensitivity and specificity of DNA FCM were thus 59.25% and 98.63%, respectively. There was a statistically significant difference (Student's unpaired t-test, P < 0.05) between cytologically malignant cases and control benign cases in all the nuclear morphometric parameters except for nuclear roundness. There was, however, no statistically significant difference of nuclear morphometric parameters between cytologically benign vs. DNA aneuploid cases and control benign cases. DNA FCM is a useful adjunct for routine diagnostic cytology. Visual diagnostic cytology and morphometric digital microscopy miss some cases of malignancy which can be detected by DNA flow cytometry. Diagn. Cytopathol. 2000;22:81-85.

The value of DNA flow cytometry in predicting the development of lymph node metastasis and survival in patients with locally recurrent oral squamous cell carcinoma.

DNA flow cytometry studies of squamous cell carcinoma of the head and neck have shown that patients with diploid tumors have favorable prognoses, whereas the outcomes of those with, aneuploid tumors are poor. This study was conducted to examine the importance of DNA ploidy in patients with locally recurrent oral carcinoma. DNA flow cytometry was performed on 93 primary oral carcinomas and their subsequent recurrences. Eight patients with diploidy of both the primary tumor and the recurrence never developed lymph node metastasis. The 5-year overall survival rate of this group was 87%. For 80 aneuploid primary carcinomas, recurrences developed that were also aneuploid. Only 31% of these patients were 5-year survivors (P < 0.001). Lymph node metastasis at presentation was found in 55% of this group, whereas 13% of initially lymph node negative patients presented with regional disease at second surgery. Five of 13 diploid primary tumors recurred with aneuploid cell lines. Three of these five patients died, two with regional metastasis. The 5-year survival rate of patients with aneuploid recurrences who were never afflicted with lymph node involvement (41%) was better (P < 0.05) than the 5-year survival rate of those with metastasis at presentation or at second surgery (26%). The excellent prognosis of patients with diploid primary tumors can be reestablished by treating local recurrences with radical surgery, if the surgery is performed before aneuploid cell lines have emerged. It appears that aneuploid tumor cell lines acquire unique properties that make them capable of invasion and metastasis.

Prognostic Factors in Patients with Bulky Stage IB or IIA Cervical Carcinoma Undergoing Neoadjuvant Chemotherapy and Radical Hysterectomy

All patients with bulky (≥4 cm) Stage Ib or IIa cervical carcinoma treated at Chang Gung Memorial Hospital between August 1988 and December 1991 using a strategy of neoadjuvant chemotherapy with cisplatin, vincristine, and bleomycin and radical hysterectomy were reviewed. Fifty-nine evaluable patients received 1 to 3 courses of chemotherapy, and 51 underwent subsequent hysterectomy. The remaining 8 patients, not completing planned surgery, were treated with definitive radiotherapy. The overall clinical response rate was 81.4% (48/59) with 18.6% complete response. Clinical response to chemotherapy was not different by stage, histologic type, tumor size, level of squamous cell carcinoma antigen, or DNA ploidy. However, tumors with DNA indices (DI) greater than 1.3 were associated with higher clinical response rates than tumors with DI ≤ 1.3 (P= 0.043). Histologically proven pelvic node metastases was noted in 18.5% (10/54) who had laparotomy. Concomitant pregnancy and more than one node metastases had significant adverse influence on recurrence and death. The 5-year survival rate of those patients who received hysterectomy was 80.3%, while only 1 of the 8 patients without hysterectomy survived. Of the 7 patients received hysterectomy despite clinical poor response, only 2 had node metastases and 3 died, whereas all the 4 patients deterred hysterectomy for poor response died. This study demonstrates the value of DNA flow cytometry in predicting chemosensitivity. However, with a DI cutoff at 1.3, only 29.2% patients could be selected. Further studies are necessary to find additional indicators that predict histological response to select better candidates for this approach and to determine optimal adjunctive treatment in case that poor prognostic features are found.

Prognostic evaluation of DNA flow cytometric and histopathologic parameters of colorectal cancer

The clinical value of DNA flow cytometry of colorectal cancer is unclear. The purpose of this retrospective study was to evaluate the relationship between tumor flow cytometry, histopathologic parameters, and survival. Flow cytometry was performed on paraffin embedded specimens from 653 patients who had surgery from 1980 to 1983. Aneuploidy was associated with distal tumor, perineural invasion, desmoplastic reaction, and failure to secrete mucin. TNM Stage I tumors were more frequently diploid than were more advanced tumors (71% vs. 41%). An abnormal DNA content had a marginal impact on survival as evaluated by univariate analysis (69% vs. 61% 10-year survival rate, P = 0.06). Multivariate analysis revealed that significant predictors of outcome were lymph node metastasis (95% confidence interval of relative risks of death from recurrent disease, 1.50-2.92), rectal cancer (1.22-2.19), absence of lymphocytic infiltration (1.20-2.17), invasion through bowel wall (1.17-3.13), lymphatic vessel invasion outside bowel wall (1.05-2.69), perineural invasion (1.15-3.19), and male gender (1.00-1.79). These findings suggest that ploidy is associated with some histopathologic parameters, but flow cytometry does not correlate with long term survival of patients with colorectal carcinoma.

L'analisi del DNA mediante citometria a flusso nei tumori superficiali della vescica: DNA analysis through flow cytometry in superficial bladder tumours

The aim of this work is to try to characterize the prognostic value of DNA flow cytometry in superficial bladder tumours and to try to indicate this investigation in clinical practice. The Authors review results reported in literature and those relative to their experience. Data investigation shows that this test has prognostic significance because it is correlated with the stage and grade of the tumour. The correlation with the clinical development of the tumour is not clear in the Authors’ experience. The Authors think that flow cytometry does not have a precise role in clinical practice.

Flow cytometric measurement of proliferation-associated nuclear antigen P105 and DNA content in immuno-proliferative small intestinal disease (IPSID).

Immunoproliferative small intestinal disease (IPSID), most common in Mediterranean countries, is characterized by lymphomatous infiltration of the small intestine and is usually associated with the synthesis of anomalous immunoglobulin alpha heavy chains. Flow cytometric analysis of DNA content, S phase fraction, and quantitative analysis of the proliferation-associated nuclear antigen, P105, were performed in 23 patients with IPSID to determine if they could be used as prognostic indicators in this disease. Eighteen patients had low-grade, two had intermediate-grade, and three had high-grade lymphoma. Eight patients had clinical stage IE disease, 12 had stage IIE, and three had stage IIIE disease. Eleven patients survived > 5 yr (good prognosis), four survived between 2-5 yr (intermediate prognosis), and eight survived 2 yr or less (poor prognosis). The S phase fraction of patients with poor prognosis was significantly higher than those with intermediate or good prognosis (P < 0.004). Flow cytometric evaluation of S phase fraction may offer important prognostic information in patients with IPSID and could be useful in the clinical management of patients with this highly variable clinical syndrome. Further studies evaluating the value of DNA flow cytometry in larger groups of patients with IPSID are warranted.

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.

Prognostic value of DNA flow cytometry in the locally recurrent, conservatively treated breast cancer patient.

This study attempted to determine the prognostic value of DNA flow cytometry in the treatment of patients with locally recurrent, conservatively treated breast cancer. Of 433 patients with clinical stage I and II breast cancer treated with conservative surgery and radiotherapy at Yale-New Haven Hospital before January 1985, 50 patients experienced an ipsilateral breast relapse as a first site of treatment failure. Using standard flow-cytometric techniques, DNA ploidy, DNA index, and S-phase fraction (SPF) were measured for 38 of the 50 (76%) paraffin-embedded specimens available for analysis. At a median postrecurrence follow-up of 5.8 years, the 5-year and disease-free survival rates following ipsilateral breast treatment failure were 48% and 54%, respectively. Sixty-three percent of the recurrent tumors were DNA diploid and 37% were aneuploid. Both DNA ploidy and SPF were statistically significant prognostic indicators for 5-year survival and disease-free survival after local recurrence. The 5-year survival rate of the DNA diploid population was 64%, compared with 15% in the aneuploid population (P < .02). Patients with low SPF (< 12%) experienced an 83% 5-year survival rate, compared with a 24% 5-year survival rate in patients with high SPF (> or = 12%) (P < .03). Ploidy and SPF were combined to define the categories of favorable (diploid, low SPF) and unfavorable (diploid, high SPF or any aneuploid subgroups). Patients in the favorable category experienced an 89% 5-year postrecurrence survival rate and a 100% disease-free survival rate, whereas patients in the unfavorable category had a 24% 5-year survival rate and a 32% disease-free survival rate (P < .01). The flow cytometry as a factor correlated with other clinical parameters previously shown to be of prognostic significance in this patient population. In a multivariate analysis, flow cytometry was a statistically significant and independent prognostic factor for disease-free survival following local recurrence. DNA ploidy and SPF as measured by currently available flow-cytometric techniques show promise as a tool in determining prognosis for the patient with locally recurrent breast cancer. Implications of these findings with respect to issues of adjuvant systemic therapy at the time of local recurrence are discussed.

Improving the prognostic value of DNA flow cytometry in metastatic melanoma by combinig ploidy and S-phase fraction

The prognostic value of cellular DNA content of melanoma metastases was investigated in tumours from 114 consecutive patients referred to the Helsinki University Central Hospital Melanoma Team. Thirty-six percent of the tumours were diploid and 64% aneuploid. For 91 patients the S-phase fraction was calculable. Tumour ploidy and S-phase fraction (SPF) were shown by multivariate Cox model analysis to be independent prognostic variables and major determinants of survival after first recurrence. Patients with either aneuploid or low SPF tumours survived longer than did those with diploid or high SPF tumours. By combining DNA ploidy and SPF, three types of DNA histograms could be defined, associated with favourable, intermediate and poor prognosis. Patients with aneuploid, low SPF metastases showed a median survival of 57 months, whereas the high-risk group with diploid, high SPF metastases survived only 13 months. When ploidy, SPF, age, sex, TNM stage and duration of disease-free survival were analysed as covariates the division of flow cytometry histograms into these three types resulted in the most significant prognostic factor (p < 0.001) in the Cox multivariate analysis.

Prognostic relevance of DNA flow cytometry in oligodendroglioma.

In a retrospective study of 85 cases, the prognostic value of DNA flow cytometry in oligodendrogliomas was evaluated. Paraffin-embedded material was processed for flow cytometry, and the survival rates of the patients with DNA diploid, aneuploid, and tetraploid tumors were compared using analysis of variance. In addition, the mitotic index was correlated with the results of flow cytometry. Finally, the results of flow cytometry, histopathologic grading, and counting mitoses were tested for dependency. Thirty-one percent of the tumors were diploid, 39% were tetraploid, and 31% were aneuploid. The results of the DNA flow cytometry did not correlate with the survival times (P = 0.798) or with tumor degree. In contrast, the number of mitoses (P less than 0.05), and the grades of the grading system of Smith (P less than 0.003) had relevance for the prognosis. No correlation between flow cytometry, histopathologic grading, and mitotic index was found. It is concluded that flow cytometry has no value in predicting the biologic behavior of oligodendrogliomas, whereas the number of mitoses is a valuable prognostic parameter and thus is considered to be incorporated into the grading system for oligodendrogliomas.

La cytométrie en flux: intérêt pronostique dans les cancers mammaires. Étude de 685 cas

The prognostic value of DNA flow cytometry has been evaluated by a prospective study of 685 cases of breast cancers. All patients were initially surgically treated. Multivariate analysis on the Cox model showed that flow cytometry classification appears to have major pronostic importance, before histopronostic classification and Progesterone Receptor status. Moreover, determination of proportion of cells DNA-synthesis (S %) phase allows definition of subgroups with poor prognosis within population generally expected to have a favorable outcome.

Pathology of the uterine body

During the past year, evidence has compiled to suggest that uterine carcinosarcomas are, in fact, metaplastic carcinomas and that mitotic counts are of no value in distinguishing between high- and low-grade endometrial stromal sarcomas in stage I cases. The pathologic and biologic behaviour of uterine müllerian adenosarcomas has been comprehensively reviewed. Prognostic factors have been identified in a large series of surgically staged endometrioid adenocarcinomas of the endometrium, while the sinister nature of the serous papillary and clear cell carcinomas of the endometrium have been confirmed. It has been claimed that a papillary pattern of growth is an independent prognostic factor in endometrial neoplasia and shown that the accuracy of macroscopic estimation of depth of myometrial invasion decreases with increasing tumor grade. The value of peritoneal cytologic examination in early stage endometrial adenocarcinoma has been questioned, as has the value of DNA flow cytometry in endometrial neoplasia. The prognostic significance of steroid receptor status has been confirmed and studies of oncogene amplification are beginning to yield prognostically useful information. Evidence is also beginning to emerge concerning chromosomal abnormalities in many women with endometrial adenocarcinoma.

DNA ploidy of primary breast cancer and local recurrence after breast-conserving therapy

The value of DNA-flow cytometry and clinico-pathological prognostic factors for the prediction of local recurrences after breast-conserving therapy (BCT) were evaluated in a retrospective study. Thirty-one patients with a local recurrence were compared with 31 matched patients without a local recurrence. Morphology and DNA-indices of the local recurrences and their corresponding primary tumours were compared. Ductal carcinoma in situ was present significantly more often in the group with a primary recurring tumour, than in the matched group (P less than 0.001), and the same holds for lobular carcinoma (n = 5). Half of the tumours that recurred had macroscopically positive surgical margins compared to about one-fourth of the matched group. Fifty-six per cent of the DNA-aneuploid stemlines in cases with local recurrence were present in the corresponding primary tumour as well (confidence limits 45%-75%), an indication that the majority of local recurrences are true recurrences and not independently developed tumours. The lack of similarity of DNA stemlines between some primary DNA-aneuploid tumours and their local recurrences indicates that these tumours had developed independently. The percentage of DNA-aneuploid cases in the group with local recurrence (89%) did not differ significantly from that in the matched group (70%). However, the findings suggest a selective recurrence of DNA-diploid stemlines. This might indicate increased resistance of DNA-diploid tumour cells to radiotherapy as compared with the resistance level in DNA-aneuploid cells.

Prognostic value of DNA flow cytometry in paragangliomas of the carotid body

A carotid body tumor is a paraganglioma of the carotid bifurcation. Histologic appearance does not correlate with the malignant potential of the lesion, and thus a reliable prognostic marker for these tumors is lacking. To determine whether flow cytometric analysis of paraffin block specimens by DNA index and synthetic phase fraction (SPF) would be of prognostic value, a retrospective chart review of 14 patients with carotid body tumors was performed. Three of 14 tumors were aneuploid and were the only tumors with SPF greater than 15%. One of 3 patients (33%) with an aneuploid tumor (SPF = 22%) developed a local recurrence; no patient with a diploid tumor developed a recurrence. Two of 3 (67%) patients with aneuploid tumors (SPF = 18%) but only 1 of 11 (9%) with a diploid tumor were symptomatic preoperatively (P = 0.03). DNA index and SPF may help select a subgroup of patients with more aggressive tumors who are at increased risk for recurrence and therefore require closer follow-up.

The prognostic value of dna flow cytometry in the locally recurrent conservatively treated breast cancer patient

The prognostic value of dna flow cytometry in the locally recurrent conservatively treated breast cancer patient

The value of DNA flow cytometry in low grade stromal sarcoma

The value of DNA flow cytometry in low grade stromal sarcoma