Prognostic Value Of DNA Ploidy Research Articles

P1.24-06 Prognostic Value of DNA Ploidy and Nucleotyping in Early-stage Non-small Cell Lung Cancer

P1.24-06 Prognostic Value of DNA Ploidy and Nucleotyping in Early-stage Non-small Cell Lung Cancer

Prognostic value of a novel biomarker combining DNA ploidy and tumor burden score for initially resectable liver metastases from patients with colorectal cancer

BackgroundColorectal cancer liver metastases (CRLM) has not been identified as a unified disease entity due to the differences in the severity of metastatic disease and tumor aggressiveness. A screen for specific prognostic risk subgroups is urgently needed. The current study aimed to investigate the prognostic value of DNA ploidy, stroma fraction and nucleotyping of initially resectable liver metastases from patients with CRLM.MethodsOne hundred thirty-nine consecutive patients with initially resectable CRLM who underwent curative liver resection from 2006 to 2018 at Sun Yat-sen University Cancer Center were selected for analysis. DNA ploidy, stroma fraction and nucleotyping of liver metastases were evaluated using automated digital imaging systems. Recurrence-free survival (RFS) and overall survival (OS) were analyzed using the Kaplan-Meier method and Cox regression models.ResultsDNA ploidy was identified as an independent prognostic factor for RFS (HR, 2.082; 95% CI 1.053–4.115; P = 0.035) in the multivariate analysis, while stroma-tumor fraction and nucleotyping were not significant prognostic factors. A significant difference in 3-year RFS was observed among the low-, moderate- and high-risk groups stratified by a novel parameter combined with the tumor burden score (TBS) and DNA ploidy (72.5% vs. 63.2% vs. 37.3%, P = 0.007). The high-risk group who received adjuvant chemotherapy had a significantly better 3-year RFS rate than those without adjuvant chemotherapy (46.7% vs. 24.8%; P = 0.034).ConclusionsOur study showed that DNA ploidy of liver metastases is an independent prognostic factor for patients with initially resectable CRLM after liver resection. The combination of DNA ploidy and TBS may help to stratify patients into different recurrence risk groups and may guide postoperative treatment among the patients.

Prognostic Value of DNA Ploidy By Flow Cytometry for Pediatric B-Cell Acute Lymphoblastic Leukemia: A Peruvian Cohort Study

Background: B-cell Acute Lymphoblastic Leukemia (B-ALL) represents an aggressive malignancy but highly curable in children. Currently, pediatric collaborative clinical trials have reported survival rates that exceed 90%, and DNA ploidy by flow cytometry (FC) has been pointed for risk stratification and prognosis in these clinical trials. However, its generalized use remains controversial as few studies reported no impact in real-world populations. We aimed to evaluate prognostic value of DNA ploidy measured by FC in a large cohort of Peruvian children with B-ALL. Methods: We evaluated prospectively DNA-ploidy by FC in bone marrow diagnostic samples from newly diagnosed children (<15 years) with B-ALL treated at Instituto Nacional de Enfermedades Neoplasicas (Lima-Peru) between 2017-2019. DNA-ploidy was evaluated using Propidium Iodide and calculated as the mean ratio between fluorescence of pathologic B blasts and normal marrow cells. Ploidy categories were established based on previous reports of DNA-index(DI): Diploid + Low-Hyperdiploidy (DLH; DI: 0.95 - 1.15), hypo-diploid (HD; DI<0.95) and High-hyperdiploidy (HH; DI>1.15), and recalculated using maximally selected rank statistics. Samples were analyzed in a FacsCanto II flow cytometer (BD) and Infinicyt software (Cytognos). All patients received BFM-2009 protocol and had minimal residual disease evaluation at the end of IA and IB induction. Minimal residual disease (MRD) was evaluated with FC using a detection threshold of 0.0025% and considering positive ≥0.01%. Survival curves (event-free and overall survival) were estimated using the Kaplan-Meier method and compared with the Log-rank test. Results: A total of 192 children were included (2 HD, 141 DLH and 49 HH cases according to DNA ploidy by FC). Clinical characteristics and outcomes are shown in Table 1. Median age at diagnosis was 5 years (Range:1-14), 10 years for HD, 6 and 3 years for DLH and HH, respectively (p=0.002). F/M ratio was 1:1.3 for all cases, but 1:2 in HH group. Most karyotypes (62%) had unsatisfactory or poor-quality result, 29% were considered normal, and only 9 hyperdiploidy and 2 hypodiploidy cases were detected by conventional karyotyping. Regarding genetics, 21 TEL/AML, 19 E2A/PBX1 and 9 BCR/ABL cases were detected by multiplex-PCR and most balanced alterations had DLH subtype and only one HD case had TEL/AML. MRD positivity after Induction IA was 35% without difference between groups (50% HD, 36% DLH and 31% HH, p=0.77), however after Induction IB, MRD was positive in 50% of HD, 12% of DLH and 5% of HH (p=0.048). At eight-teen months of follow-up, one relapse was seen in HD cases, and 11% DLH and 10% in HH. Median EFS and OS was not reached, however one-year EFS and OS were 86% for both without significant differences between groups. Multivariate analysis showed that MRD positivity remains as the principal independent prognostic factor and ploidy by DNA did not show any impact in terms of MRD, EFS and OS. Conclusion: High-Hyperdiploidy by DNA ploidy was associated to better MRD negativity rate after induction IB but without impact in short-term EFS and OS. DNA ploidy did not represent a prognostic factor in our study cohort, however long-term follow-up is warranted. Additionally, a better genetic risk stratification is necessary to improve outcomes in Latino high-risk population. Disclosures No relevant conflicts of interest to declare.

Prognostic value of nucleotyping, DNA ploidy and stroma in high-risk stage II colon cancer

BackgroundHeterogeneity with respect to recurrence and survival in high-risk stage II colon cancer patients still exists, and further classification is urgently required. This study aimed to ascertain the prognostic value of DNA ploidy, stroma-tumour fraction and nucleotyping in the prognosis of high-risk stage II colon cancer.MethodsA total of 188 high-risk stage II colon cancer patients received radical surgery in Peking University Cancer Hospital, from 2009 to 2015. Status of mismatch repair proteins in tumours was analysed using immunohistochemistry. DNA ploidy, stroma-tumour fraction and nucleotyping were estimated by automated digital imaging systems.ResultsNucleotyping and DNA ploidy were significant prognostic factors, while stroma-tumour fraction were not significantly prognostic in the univariate analysis. In the multivariable model, the dominant contributory factor of disease-free survival was chromatin heterogeneous vs. chromatin homogeneous [HR 3.309 (95% CI: 1.668–6.564), P = 0.001].ConclusionsOur study indicates that nucleotyping is an independent prognostic factor in high-risk stage II colon cancer. Therefore, it may help subdivide patients into different subgroups and give them different strategies for follow-up and treatment in the future.

Prognostic value of DNA ploidy and automated assessment of stroma fraction in prostate cancer.

The combination of DNA ploidy and automatically estimated stroma fraction has been shown to correlate with recurrence and cancer death in colorectal cancer. We aimed to extend this observation and evaluate the prognostic importance of this combined marker in prostate cancer. DNA ploidy status was determined by image cytometry and the stroma fraction was estimated automatically on hematoxylin and eosin stained sections in three tumor samples from each patient to account for tumor heterogeneity. The optimal threshold for low (≤56%) and high (>56%) stroma fraction was identified in a discovery cohort (n = 253). The combined marker was validated in an independent patient cohort (n = 259) with biochemical recurrence as endpoint. The combined marker predicted biochemical recurrence independently in the validation cohort. Multivariable analysis showed that the highest risk of recurrence was observed for patients with samples that had both non-diploid ploidy status and a high stroma fraction (hazard ratio: 2.51, 95% confidence interval: 1.18-5.34). In conclusion, we suggest the combination of DNA ploidy and automatically estimated stroma fraction as a prognostic marker for the risk stratification of prostate cancer patients. It may also be a potential generic marker as concurrent results have been described in colorectal cancer.

Biomarker-based prediction of inflammatory bowel disease-related colorectal cancer: a case–control study

BackgroundRegular colonoscopic surveillance for detection of dysplasia is recommended in longstanding inflammatory bowel disease (IBD), however, its sensitivity is disputed. Screening accuracy may increase by using a biomarker-based surveillance strategy.MethodsA case-control study was performed to determine the prognostic value of DNA ploidy and p53 in IBD-related neoplasia. Cases with IBD-related colorectal cancer (CRC), detected in our surveillance program between 1985-2008, were selected and matched with two controls, for age, gender, disease characteristics, interval of follow-up, PSC, and previous surgery. Biopsies were assessed for DNA ploidy, p53, grade of inflammation and neoplasia. Progression to neoplasia was analyzed with Cox regression analysis, adjusting for potentially confounding variables.ResultsAdjusting for age, we found statistically significant Hazard ratios (HR) between development of CRC, and low grade dysplasia (HR5.5; 95%CI 2.6-11.5), abnormal DNA ploidy (DNA index (DI) 1.06-1.34, HR4.7; 95%CI 2.9-7.8 and DI>1.34, HR6.6; 95%CI 3.7-11.7) and p53 immunopositivity (HR3.0; 95%CI 1.9-4.7) over time. When adjusting for all confounders, abnormal DNA ploidy (DI 1.06-1.34, HR4.7; 95%CI 2.7-7.9 and DI>1.34, HR5.0; 95%CI 2.5-10.0) and p53 immunopositivity (HR1.7; 95%CI 1.0-3.1) remained statistically significant predictive of neoplasia. ConclusionIn longstanding IBD, abnormal DNA ploidy and p53 immunopositivity are important risk factors of developing CRC. The yield of surveillance may potentially increase by adding these biomarkers to the routine assessment of biopsies.

The prognostic value of DNA ploidy in a total population of uterine sarcomas

Background: The diagnosis of uterine sarcoma is associated with poor outcome for the patient and there is a need for reliable prognostic markers. Most previous studies on the prognostic value of DNA ploidy include few uterine sarcomas and report conflicting results.Materials and methods: We examined the prognostic value of DNA ploidy and its association with clinicopathological parameters and crude survival in a total population of 354 sarcoma.Results: In univariate analyses, we observed significantly better crude survival for endometrial stromal sarcomas (ESS) and adenosarcoma (AS) patients with diploid as compared with nondiploid tumors, but not for patients with leiomyosarcomas (LMS). In Cox multivariate analyses, DNA ploidy was the only significant predictor of survival for patients with AS. In LMS, mitotic index (MI), tumor size, tumor extent and tumor margins, whereas for ESS, MI, tumor extent and tumor necrosis obtained independent significance of survival. DNA ploidy was a significant predictor of survival for LMS patients in Cox regression analyses when excluding MI.Conclusion: DNA ploidy might be useful as a prognostic marker in patients with LMS and AS.

Prognostic Value of DNA Ploidy, bcl-2 and p53 in Localized Prostate Adenocarcinoma Incidentally Discovered at Transurethral Prostatectomy

Discovery of prostatic adenocarcinoma limited to transurethral resection material generates a treatment dilemma. We investigated the usefulness of parameters shown to be associated with prognosis in prostate cancer (p53 and bcl-2 immuno-expression, DNA cell cycle analysis and Gleason score) to stratify these incidentally identified tumors to guide clinical decision making. Paraffin embedded tissues from transurethral prostate resection specimens containing T1a prostate adenocarcinoma from 44 patients who underwent resection between 1980 and 1990 were immunostained for p53 and bcl-2, and subjected to flow cytometry to determine DNA ploidy. Gleason score was determined by 2 pathologists independently. Statistical relationships among these 4 variables, tumor progression and cancer specific survival were analyzed. Six of 44 patients in the study population had cancer progression. Time to clinical progression was 4.5 years (range 7 months to 11 years). Most tumors stained negative for p53 and bcl-2. Only 2 tumors studied were aneuploid and neither of these 2 patients had cancer progression. Only Gleason score was a significant predictor of cancer progression on univariate and multivariate Cox regression analysis (p = 0.045 and 0.046, respectively). No tumor characteristics correlated with time to disease progression, including p53 and bcl-2 immuno-expression, and Gleason score (p = 0.182, 0.563 and 0.346, respectively). Positive immunostaining for p53 and bcl-2 did not occur together in the same tumor in significant fashion (p = 0.334), nor did either significantly occur more with aneuploidy (p = 0.237 and 0.307 respectively). For T1a prostate cancer incidentally detected on transurethral prostate resection p53 and bcl-2 immuno-expression, and DNA ploidy do not predict survival or disease progression.

Prognostic value of DNA ploidy: 5-year follow-up of patients with resectable squamous cell carcinoma (SCC) of the lung

This study was designed to show the relation between DNA-ploidy in patients with resectable lung cancers and their 5-year survival rate. The results are compared with those from our 2-year follow-up study of the same group of patients published in 2000. The group of 80 patients with SCC who underwent lung resection between 1995 and 96 were re-analyzed. For the statistical analysis the hazard Cox model and an exponential multiple regression model were used. The survival curves were drawn using the Kaplan-Meier method. DNA-aneuploidy was found in 45% of cancer tumors. There was no statistically significant correlation between aneuploidy and gender, age, cancer staging or grading. In the 3-year follow-up the survival rate in patients with aneuploid type tumors was significantly lower than in those with the diploid type. However, this difference was not found after 5 years of follow-up. Tumor ploidy was an independent prognostic factor only in patients between 55 and 60 years of age. The mortality rate in patients with aneuploid tumors was mainly the result of distant metastases while, in patients with diploid tumors, local recurrence was the main reason for death. In the first three years after surgical resection patients with aneuploid tumors are at higher risk of distant metastases than patients with the diploid type. Tumor ploidy can be recognized as an independent prognostic factor in younger (55-60) patients. Aneuploidy promoted the occurrence of early distant metastases while the diploid type was associated with late (after 3 years) local tumor recurrence.

Prognostic value of DNA ploidy in primary gastric leiomyosarcoma.

The prognostic value of DNA ploidy was evaluated in 25 patients with primary gastric leiomyosarcoma, using paraffin-embedded archival specimens analysed by DNA flow cytometry. Ploidy patterns were diploid in ten tumours and non-diploid in 15. They did not correlate with clinicopathological features of the lesions such as tumour size and grade or mitotic index. A non-diploid pattern was associated with a significantly reduced patient survival rate (P < 0.05), as was higher tumour grade (P < 0.01). Neither tumour size nor mitotic index correlated with patient survival.

Node negative breast cancer: The prognostic value of DNA ploidy for long-term survival

The DNA content of breast tumours from 170 patients who presented between 1978 and 1980 was measured by flow cytometry. The relationship between tumour ploidy and disease outcome was assessed and its association with other prognostic factors evaluated. Compared with those with diploid tumours, patients with aneuploid tumours had significantly earlier relapse and shorter survival (P less than 0.0001). Tumour ploidy was strongly related to grade (P less than 0.001), but there was no significant association between DNA ploidy and c-erb-B-2 expression, lymph node status or tumour size. In lymph node negative and c-erb-B-2 negative patients, aneuploid tumours were associated with a poorer prognosis (P less than 0.001) than diploid tumours. Multivariate analysis showed that tumour ploidy gave independent information on disease free and overall survival. Tumour ploidy may be used as an independent prognostic variable in patients with breast cancer and it may be helpful in defining patients within the node negative or c-erb-B-2 negative groups likely to have a poor outcome who might benefit from adjuvant treatment.

INTRACAPSULAR CLEAR CELL RENAL CARCINOMA: PLOIDY STATUS IMPROVES THE PROGNOSTIC VALUE OF THE 2002 TNM CLASSIFICATION

The TNM classification has been revised for the 2002 edition of the UICC publication to better stratify patients with intracapsular renal cell carcinoma (RCC) but few studies have been published to date to validate this new classification. Moreover, additional prognostic factors seem to be necessary to improve the prediction of intracapsular tumor aggressiveness and the definition of patient subgroups at high risk for metastases. We report the long-term results of the new TNM scheme. We evaluated the impact of DNA content, S-phase and MIB-1 (Dako, Glostrup, Denmark) score. A total of 136 patients with intracapsular clear cell RCC and a mean followup of 74 months were reclassified. Tumor specific survival (TSS) was compared with nuclear grade (NG), DNA content and proliferative status (S-phase fraction and MIB-1 score). TSS was 92%, 81.1% and 40.1% for pT1a, pT1b and pT2, respectively (p <0.05). TSS according to DNA ploidy status (diploid vs aneuploid) was pT1a-95.2% vs 68.6% (p <0.05), pT1b-90% vs 46.7% (p <0.05) and pT2-49.2% vs 25% (p not significant). DNA ploidy was also significantly associated with survival when adjusted for NG. There was no significant association between TSS and MIB-1 score or tumor S-phase fraction. The 2002 TNM classification is a useful prognostic factor for evaluating organ confined RCC of the clear cell subtype. Evaluation of the DNA content in clear cell RCC appears to significantly improve the predictive value of the TNM staging system, especially in the pT1a and pT1b categories. Fuhrman NG alone or combined should be routinely used in such patients.

Prognostic Biomarkers in Renal Cell Carcinoma: Relevance of DNA Ploidy in Predicting Disease-Related Survival

To investigate the prognostic value of DNA ploidy, Ki-67 index and p53 expression in relation to disease-related survival in a consecutive series of patients with renal cell carcinoma (RCC). The study group consisted of 64 RCC patients treated by radical nephrectomy. Histological type, pathological staging and nuclear anaplasia were assessed according to the WHO classification, TNM system and Fuhrman grading criteria, respectively. Ploidy was determined by DNA flow cytometry using two sampling methods (frozen vs paraffin-embedded tissue). Ki-67 and p53 were evaluated by immunohistochemistry techniques using two cutoff points (10% vs mean value) for staining interpretation. Kaplan-Meier and Cox regression analyses were used for prognostic evaluation. Thirty-one tumors (48.4%) showed DNA diploidy and 33 (51.6%) were DNA aneuploid. Concordance between both ploidy measurement methods was found in 85.5% of cases (p=0.0455). The mean values for Ki-67 and p53 immunostaining were 3.65% (0-23.5%) and 5.90% (0-55.9%), respectively. DNA ploidy significantly correlated with staging, tumor size (pT), nuclear grading, and Ki-67 (mean value cutoff). Ki-67 (10% cutoff) correlated with staging and pT, while p53 (mean value cutoff) was associated with Ki-67 (mean value cutoff). There were significant differences between survival curves for pathological stage, pT, nuclear grade, ploidy, Ki-67 (both cutoffs), and p53 (10% cutoff). By univariate regression analysis, stage III and stage IV, pT3, aneuploidy, high Ki-67 (both cutoffs), and p53 overexpression (10% cutoff) showed significant correlations with worse disease-related survival. In addition, DNA aneuploidy significantly correlated with poor prognosis within stages I/II (p=0.0355) and stages III/IV (p=0.0138) of the disease. The results indicate that DNA ploidy has relevant prognostic value in RCC, adding useful information to the classic histopathological indicators of clinical outcome.

Prognostic Significance of DNA Ploidy and Proliferative Index (MIB-1 Index) in Childhood Rhabdomyosarcoma

The prognostic value of DNA ploidy and proliferative index (PI) are well established in many cancers, but their significance in childhood rhabdomyosarcoma (RMS) is unclear. We studied the DNA content and PI of 45 cases of childhood RMS obtained retrospectively. DNA histograms were hyperdiploid in 30 cases (67%), diploid in 6 (13%), tetraploid in 5 (11%), polyploid in 3 (7%), and nonclassifiable in 1 (2%). The 5-year overall survival rate by ploidy was 60% (3/5) in tetraploid, 57% (17/30) in hyperdiploid, and 0% in diploid and polyploid cases (P = .000002). The 5-year overall survival by a PI less than or greater than 19% was 62% (13/21) and 21% (5/24), respectively (P = .006). In multivariate analysis, DNA ploidy (P = .001) was an important independent prognostic factor. DNA content in childhood RMS is an important variable in predicting prognosis. DNA hyperdiploid and tetraploid rhabdomyosarcomas had a favorable prognosis, while DNA diploid and polyploid tumors had a poor prognosis.

Prognostic significance of DNA ploidy and proliferative index (MIB-1 index) in childhood rhabdomyosarcoma.

The prognostic value of DNA ploidy and proliferative index (PI) are well established in many cancers, but their significance in childhood rhabdomyosarcoma (RMS) is unclear. We studied the DNA content and PI of 45 cases of childhood RMS obtained retrospectively. DNA histograms were hyperdiploid in 30 cases (67%), diploid in 6 (13%), tetraploid in 5 (11%), polyploid in 3 (7%), and nonclassifiable in 1 (2%). The 5-year overall survival rate by ploidy was 60% (3/5) in tetraploid, 57% (17/30) in hyperdiploid, and 0% in diploid and polyploid cases (P = .000002). The 5-year overall survival by a PI less than or greater than 19% was 62% (13/21) and 21% (5/24), respectively (P = .006). In multivariate analysis, DNA ploidy (P = .001) was an important independent prognostic factor. DNA content in childhood RMS is an important variable in predicting prognosis. DNA hyperdiploid and tetraploid rhabdomyosarcomas had a favorable prognosis, while DNA diploid and polyploid tumors had a poor prognosis.

Valor pronóstico de la ploidía del adn y la morfometría nuclear en el cáncer de próstata metastásico

Prognostic value of dna ploidy and nuclear morphometry in metastatic prostate cancer PurposeTo assess the prognostic value of DNA ploidy and nuclear morphometry in metastatic prostate cancer after androgenic deprivation treatment. MethodsFifty four patients with prostate cancer and bone metastases who had undergone androgenic suppression treatment were retrospectively studied. The deoxyribonucleic acid (DNA) content was analysed by flow cytometry. Nuclear morphometry characterized 14 nuclear descriptors. The study also included age, Gleason score, T classification, haematocrite, serum albumin, serum alkaline phosphatase, serum prostatic acid phosphatase and the amount of metastatic foci detected during radioisotope bone scan. Univariate survival analyses were performed and Cox’s proportional hazards model was used to identify significant prognostic factors. To assess how the experimental factors improve the capacity of the classical factors for predicting the patients who reach median survival, logistic regression multivariate analysis was performed for the classical prognostic factors only and after added experimental variables (DNA content and Nuclear Area). ResultsThe univariate survival analyses assigned a prognostic value to T category, level of albumin, alkaline phosphatase, Gleason score, bone scan, DNA ploidy and mean nuclear area. In the case of the Cox regression model only Gleason score, bone scan, mean nuclear area and DNA ploidy provided independent prognostic information. In logistic regression for classic prognostic factors only Gleason score is significant (sensibility 89,3%, specificity 64%). However, when the experimental factors are added, in addition to Gleason score, radioisotope bone scan and DNA ploidy are of prognostic value (sensibility 90% and specificity 72%). ConclusionsThe study of DNA content and nuclear morphometry in the primitive tumor provides independent prognostic information in survival analysis for patients with metastatic prostate cancer. However, there is limited improvement with respect to the classical factors in predicting survival. This questions its utility in the daily clinical usage.

Flow cytometric S‐phase fraction as a complementary biological parameter for the cytological grading of non‐Hodgkin's lymphoma

Fine-needle aspiration cytology (FNAC) is a technique that can overcome tissue-sampling disaggregation problems related to DNA flow cytometry analysis. The aim of this study, with long-term follow-up (median, 72 mo), was to investigate the prognostic value of DNA ploidy and S-phase fraction (SPF) in patients with non-Hodgkin's lymphoma (NHL), and additionally, the relevance of SPF in the grading of NHLs, using FNAC. The series comprised 76 patients with NHL (32 indolent and 44 aggressive tumors, including 14 Burkitt lymphomas) and 30 patients with reactive lymph node enlargement used as a control group. DNA flow cytometry was performed on fresh samples obtained by FNAC. NHL grading was done according to the updated Kiel classification. The 5-yr overall survival of patients with NHL was determined using the Kaplan-Meier method. All samples of the control group and 81.6% of the NHLs showed a DNA diploid pattern. Fourteen cases (18.4%) were DNA aneuploid with bimodal distribution: slight hyperdiploidy and near-tetraploidy. Despite the higher incidence of aneuploidy in aggressive than in indolent tumors (22.7% vs. 12.5%), no correlation between DNA ploidy and NHL grading was observed. In contrast, SPF revealed a strong correlation with grading (P=0.0001). The mean SPF values varied from 6.5% in indolent NHLs, to 20.4% in aggressive not-otherwise-specified (NOS) NHLs, and to 35.3% in Burkitt lymphomas. Nearly all aggressive NHLs had an SPF >15%, while the vast majority of indolent NHLs showed an SPF <10%. The mean SPF value in the reactive node group was 6.6%. NHL grading significantly was correlated to survival (P=0.004) only if the Burkitt lymphomas, which showed the best prognosis, were analyzed as an independent group. There was a trend that did not reach statistical significance (P=0.072) for a worse clinical outcome of patients with aneuploid tumors. When mean SPF values were used as cutoff points to divide both indolent NHLs and aggressive NOS NHLs into two proliferative subgroups, no differences in relation to survival were found (P=0.763 and P=0.994, respectively). Also, no proliferative difference was verified between indolent NHLs and the reactive lymph node group (P=0.223). These results show that flow cytometric SPF is a valuable complementary parameter for grading NHLs on FNAC samples, but it appears to give no additional prognostic information on subset analyses.

Prognostic value of DNA ploidy and nuclear morphometry in prostate cancer treated with androgen deprivation

Objectives. To assess the prognostic value of flow cytometry and nuclear morphometry in prostate cancer after androgen deprivation treatment. Methods. A total of 127 patients with a prostate cancer diagnosis who had undergone androgen suppression were retrospectively studied. The DNA content by flow cytometry and nuclear morphometry was studied from biopsy specimens. In the patients with Stage M0, two multivariate analyses by the Cox proportional regression model were performed to determine whether the experimental variables (DNA content and nuclear area) added independent information to the classic prognostic factors (Gleason score and stage). Using the statistical analysis results, risk groups were created. Results. T and M categories, Gleason score, DNA ploidy, and mean nuclear area proved to have prognostic value in the univariate analysis. For the group of patients free of metastasis (M0), it was possible to create low, intermediate, and high-risk groups using stage and Gleason score with statistically significant differences in survival. Multivariate analysis, combining the classic and experimental variables, selected Gleason score and DNA content as prognostic independent factors. Also, risk groups with statistically significant differences in survival were created. However, the net result of combining both kinds of factors was at least as valuable as the combination of stage and Gleason score in predicting survival. Conclusions. The determination of DNA ploidy and mean nuclear area do not add enough independent information to improve the predictive value to justify their use in this group of patients treated with hormonal therapy.

Prognostic value of DNA ploidy in 653 Japanese women with node-negative breast cancer.

The prognostic value of DNA ploidy has been extensively studied in breast cancer; however, the results remain controversial. Flow cytometry (FCM) studies of DNA ploidy on frozen sections have not yet been performed in a large series of Japanese women with node-negative breast cancer. An FCM analysis of DNA ploidy was performed on frozen sections of node-negative breast cancer from 653 Japanese women, with a mean follow-up duration of 46 months. Three hundred and twenty-four (49.6%) patients showed a diploid tumor, while 329 (50.4%) showed an aneuploid tumor. There was a significant correlation between DNA ploidy and estrogen receptor (ER) status. Patients with an aneuploid tumor had a significantly worse outcome in terms of both disease-free survival (DFS; P = 0.0116) and overall survival (OS: P = 0.0492) than those with a diploid tumor, while the same effect, in terms of DFS, was also seen in ER-positive breast cancer. Multivariate analyses indicated DNA ploidy to be an independent prognostic factor for DFS, while DNA ploidy and tumor size were found to be independent prognostic factors for OS. DNA ploidy was also shown to be an independent prognostic factor for DFS in ER-positive breast cancer. Our findings demonstrated DNA ploidy, based on FCM, to have an important prognostic value in Japanese women with node-negative breast cancer.

DNA aneuploidy and high proliferative activity but not K-ras-2 mutations as independent predictors of clinical outcome in operable gastric carcinoma: results of a 5-year Gruppo Oncologico dell'Italia Meridonale (GDIM) prospective study.

The prognostic value of DNA ploidy, S-phase fraction (SPF) and K-ras-2 mutations in gastric carcinoma (GC) has not yet been clearly defined. The aim of this study was to clarify the association between biomolecular variables, tumor characteristics, and clinical outcome in GC patients. Resected specimens from a consecutive series of 69 patients with GC who underwent potentially curative surgery were studied prospectively. DNA ploidy and SPF were assessed by flow cytometry on multiple frozen tumor samples, whereas K-ras-2 mutations were detected by polymerase chain reaction followed by single-strand conformation polymorphism. All the patients involved in this study were followed up for a mean of 95 months. DNA aneuploidy was present in 72% of the cases (50 of 69), whereas 10% of these (5 out of 50) showed multiclonality. Mutations of K-ras-2 were detected in 8% of the tumors (5 of 63). Both DNA ploidy and SPF were associated with TNM stage (American Joint Committee on Cancer [AJCC] staging system) and node status. Moreover, DNA aneuploidy was significantly related to high SPF. K-ras-2 mutations were not associated with clinicopathologic variables or flow cytometric indicators. At univariate analysis, advanced TNM stage, node involvement, diffuse histotype, depth of invasion, DNA aneuploidy, and high SPF proved to be significantly related to quicker tumor relapse and to shorter overall patient survival. With multivariate analysis, DNA aneuploidy, high SPF, and depth of invasion were related to risk of tumor relapse and patient death, whereas diffuse histotype was independently related to patient risk of tumor relapse. DNA ploidy and SPF, when associated with clinicopathologic staging, might be useful for the identification of GC patients who have different risks for death or relapse of disease.