The effect of vascular endothelial growth factor-1 expression on survival of advanced colorectal cancer patients.

Colorectal cancer is third leading cause of cancer mortality. About 60% of patients had already developed metastasis at the time of diagnosis. Vascular endothelial growth factor (VEGF) is crucial for the development of neovascularization and hence metastasis. This study aimed at investigating the relation between the expression of VEGF in biopsies from surgically dissected colon cancer and the survival of those patients. Biopsies were collected from 86 patients with advanced colon cancer and sections were stained by immunohistochemistry for VEGF. Patients received chemotherapy after the operation and were followed up for disease progression and survival. The clinical data were statistically analyzed with respect to the immunohistochemistry results. The survival of the patients was significantly longer in the patients for whom biopsies showed negative or weak expression of VEGF in comparison to those with moderate to high expression (p-value = 0.04). The expression of VEGF was more frequent in the patients who died as a consequence of the disease in comparison to the 10-year survivors. In conclusion, VEGF could be related to the survival of the patients with colorectal carcinoma and should be considered as a predictor of the prognosis.

Neurofibromin Expression is Associated with Aggressive Disease and Poor Outcome in Colorectal Carcinoma.

AIM: To assess the predictive and prognostic value of neurofibromin (NF) expression in colorectal carcinoma (CRC). MATERIALS AND METHODS: The present series consists of archival samples from 191 patients with stage I, II, III, or IV CRC treated between 1981 and 1990 at the Turku University Hospital (Finland). Tumor biopsies as microarray blocks were analyzed for expression of NF by immunohistochemistry. Different grading systems were tested for NF expression. RESULTS: A significant correlation between NF expression and tumor localization was found, with tumors arising in the colon showing intense NF expression more often than those arising in the rectum (p=0.014). Higher expression of NF was more common in tumors not responding to treatment (p=0.004). Tumors with multiple metastases showed higher expression of NF than those with single metastasis only (p=0.025). Furthermore, NF expression showed a borderline (p=0.068) correlation with gender; tumors of women showed higher NF expression that those of males. On the other hand, NF expression was not significantly associated with tumor recurrence, age, lymph node involvement, tumor grade and tumor stage or disease outcome. CONCLUSION: Positive NF expression in CRC is a sign of aggressive disease and poor outcome.

Proliferative activity in Libyan breast cancer with comparison to European and Central African patients.

BACKGROUND: We evaluated the relation of proliferative indices with clinicopathological features and prognosis in breast cancer (BC) of Libyan female patients. The data were compared with corresponding results in Finland and Nigeria. PATIENTS AND METHODS: Histological samples of breast cancer from 130 patients were retrospectively studied. Mitotic activity index (MAI) and standardized mitotic index (SMI) were estimated. RESULTS: There were statistically significant correlations between the proliferative indices and most clinicopathological features, with the strongest association observed for histological grade (P = 0.01 for SMI and P = 0.003 for MAI). The proliferative differences between Libyan, Nigerian, and Finnish population were prominent. The mean values of SMI and MAI in Libyan BC patients were 32.1 mitotic figures per square millimeter and 27.3 mitotic figures per 10 high-power fields, respectively. This is clearly lower than those in Nigeria but much higher than those in Finland. The differences between countries are seen in whole material and are also present in subgroups. The results indicated that mitotic activities can be reliable prognostic indicators in Libyan BCs, as they were among Finnish and Nigerian females. Univariate and multivariate analyses found at cut-offs of 19 and 44 mitosis/mm(2) of SMI were the most significant prognostic factors. CONCLUSIONS: Proliferative indices with careful estimation of the MAI and SMI could be applied as quantitative criteria for Libyan BC to separate the patients into good, moderate, and bad prognosis groups.

High cyclooxygenase-2 expression is associated with advanced stages in colorectal cancer.

BACKGROUND: Despite compelling evidence from the genetic background and clinical studies indicating that cyclooxygenase-2 (COX2) up-regulation is a key step in carcinogenesis of colorectal carcinoma (CRC), controversy regarding its role as a prognostic factor exists. However, all evidence indicates that increased COX2 activity promotes progression of CRC. This study, aimed to evaluate the expression of COX2 in CRC, and correlate it with different patient clinicopathological data, emphasizing on the role of COX2 as a prognostic factor for CRC. MATERIALS AND METHODS: In the present study, archival samples from 145 patients with stage I, II, III, or IV CRC treated during 1981-1990 at the Turku University Hospital (Finland) were used (as microarray blocks) to analyze COX2 expression by immunohistochemistry (IHC). RESULTS: Higher levels of COX2 expression were associated with higher TNM class (p<0.06), and higher Dukes' stage (p<0.045). In contrast, there was no significant correlation with age, gender, tumor grade or lymph node status. However, univariate survival analysis of metastases showed borderline association with COX2 expression in that patients with metastases with COX2-positive tumors were alive for shorter periods of time compared with patients whose tumors had no COX2 expression (p<0.023, log-rank). CONCLUSION: COX-2 expression has shown a significant correlation with tumor stage and hence is assumed to be a prognostic factor in our cohort of colorectal cancer patients.

Prognostic value of bcl-2 expression among women with breast cancer in Libya.

We studied the association of the immunohistochemical bcl-2 expression in Libyan breast cancer with clinicopathological variables and patient outcome. Histological samples from 170 previously untreated primary Libyan breast carcinoma patients were examined. In immunohistochemistry, the NCL-L-bcl-2-486 monoclonal antibody was used. Positive expression of bcl-2 was found in 106 patients (62.4 %). The bcl-2 expression was significantly associated with estrogen receptor (p<0.0001) and progesterone receptor positive tumors (p=0.002), small tumor size (p<0.0001), low tumor grade (p<0.0001), negative axillary lymph nodes (p<0.0001), early stages (p=0.001), and low risk of metastasis (p<0.0001). Positive expression was also associated with older patients (>50 years; p=0.04). Histological subtypes and family history of breast cancer did not have significant relationship with bcl-2. Patients with positive expression of bcl-2 had lower recurrence rate than bcl-2-negative patients and better survival after median follow-up of 47 months. Patients with high bcl-2 staining were associated with the best survival. The role of bcl-2 as an independent predictor of disease-specific survival was assessed in a multivariate survival (Cox) analysis, including age, hormonal status, recurrence, histological grade, and clinical stage variables. Bcl-2 (p<0.0001) and clinical stage (p=0.016) were independent predicators of disease-specific survival. For analysis of disease-free survival, the same variables were entered to the model and only bcl-2 proved to be an independent predictor (p=0.002). Patients with positive expression of bcl-2 were associated with low grade of malignancy, with lower recurrence rate, with lower rate of death, and with longer survival time. Bcl-2 is an independent predictor of breast cancer outcome, and it provides useful prognostic information in Libyan breast cancer. Thus, it could be used with classical clinicopathological factors to improve patient selection for therapy.

Loss of MUC2 expression predicts disease recurrence and poor outcome in colorectal carcinoma.

Clinical staging and histological grading after surgery have been the "gold standard" for predicting prognosis and planning for adjuvant therapy of colorectal cancer (CRC). With the recent development of molecular markers, it has become possible to characterize tumors at the molecular level. This is important for stage II and III CRCs, in which clinicopathological features do not accurately predict heterogeneity, e.g., in their tumor response to adjuvant therapy. In the present study, archival samples from 141 patients with stage I, II, III, or IV CRC treated during 1981-1990 at Turku University Hospital (Finland) were used (as microarray blocks) to analyze MUC2 expression by immunohistochemistry. Altogether, 49.7 % of all tumors were positive for MUC2. There was no significant correlation between MUC2 expression and age (P < 0.499), tumor invasion (P < 0.127), tumor staging (P < 0.470), histological grade (P < 0.706), lymph node involvement (P < 0.854), or tumor metastasis (P < 0.586). However, loss of MUC2 expression was significantly associated with disease recurrence (P < 0.031), tumor localization (P < 0.048), and with borderline significance with gender (P < 0.085). In univariate (Kaplan-Meier) survival analysis, positive MUC2 significantly predicted longer disease-free survival (DFS) and disease-specific survival (DSS) as well. However, in multivariate (Cox) survival analysis, MUC2 lost its power as an independent predictor of DFS and DSS. Our results implicate the value of MUC2 expression in predicting disease recurrence and long-term survival in CRC.

Prognostic value of proliferation markers: immunohistochemical ki-67 expression and cytometric s-phase fraction of women with breast cancer in libya.

BACKGROUND: We evaluated the association of the immunohistochemical Ki-67 expression, and S-phase fraction with clinicopathological variables and patient outcome. PATIENTS AND METHODS: Histological samples from 100 primary Libyan breast carcinoma patients were retrospectively studied with monoclonal antibody to Ki-67. S-phase fraction was determined by DNA image cytometry. RESULTS: The median Ki-67 percentage for all tumors was 27.5%, ranging from 1 to 80% and the median S-phase fraction (SPF) was 11%, ranging from 0 to 62 %. Tumors with high Ki-67 expression were found in 76% of patients and with high SPF values in 56%. Ki-67 expression was more frequent in tumors with high SPF than low SPF. High Ki-67 and high SPF were associated with advanced stages, poor differentiation of tumors, positive lymph nodes, and distant metastasis. The Ki-67 was associated with hormone receptor negative tumors. The SPF was higher in young patients (<50 years) than in older patients. In the overall population (median follow-up 49 months), patients with high Ki-67 and high SPF had shorter survival time and predicted recurrence than patients with low Ki-67 and low SPF. In a Cox multivariate analysis, high SPF (p= 0.007), hormonal status (p= 0.001) and clinical stage (p=0.005) were independent predictors of disease-specific survival. The Ki-67 (p=0.065) in borderline significance proved to be independent predictor of disease-free survival. The SPF showed more statistically significance with a high grade of malignancy and survival time than Ki-67. CONCLUSIONS: The SPF value is useful cell proliferation marker to assess tumor prognosis. These markers may reflect the aggressive behavior of Libyan breast cancer and predict of the recurrence. It is therefore important to take these markers into consideration to select a high risk subgroup of the patients for intensive treatment.

Diagnosis delay in Libyan female breast cancer.

AIMS: To study the diagnosis delay and its impact on stage of disease among women with breast cancer on Libya. METHODS: 200 women, aged 22 to 75 years with breast cancer diagnosed during 2008-2009 were interviewed about the period from the first symptoms to the final histological diagnosis of breast cancer. This period (diagnosis time) was categorized into 3 periods: <3 months, 3-6 months, and >6 months. If diagnosis time was longer than 3 months, the diagnosis was considered delayed (diagnosis delay). Consultation time was the time taken to visit the general practitioner after the first symptoms. Retrospective preclinical and clinical data were collected on a form (questionnaire) during an interview with each patient and from medical records. RESULTS: The median of diagnosis time was 7.5 months. Only 30.0% of patients were diagnosed within 3 months after symptoms. 14% of patients were diagnosed within 3-6 months and 56% within a period longer than 6 months. A number of factors predicted diagnosis delay: Symptoms were not considered serious in 27% of patients. Alternative therapy (therapy not associated with cancer) was applied in 13.0% of the patients. Fear and shame prevented the visit to the doctor in 10% and 4.5% of patients, respectively. Inappropriate reassurance that the lump was benign was an important reason for prolongation of the diagnosis time. Diagnosis delay was associated with initial breast symptom(s) that did not include a lump (p < 0.0001), with women who did not report monthly self examination (p < 0.0001), with old age (p = 0.004), with illiteracy (p = 0.009), with history of benign fibrocystic disease (p = 0.029) and with women who had used oral contraceptive pills longer than 5 years (p = 0.043). At the time of diagnosis, the clinical stage distribution was as follows: 9.0% stage I, 25.5% stage II, 54.0% stage III and 11.5% stage IV.Diagnosis delay was associated with bigger tumour size (p <0.0001), with positive lymph nodes (N2, N3; p < 0.0001), with high incidence of late clinical stages (p < 0.0001), and with metastatic disease (p < 0.0001). CONCLUSIONS: Diagnosis delay is very serious problem in Libya. Diagnosis delay was associated with complex interactions between several factors and with advanced stages. There is a need for improving breast cancer awareness and training of general practitioners to reduce breast cancer mortality by promoting early detection. The treatment guidelines should pay more attention to the early phases of breast cancer. Especially, guidelines for good practices in managing detectable of tumors are necessary.

Prognostic significance of DNA image cytometry in Libyan breast cancer.

BACKGROUND: We evaluated the relation of nuclear DNA content and clinicopathological features and prognosis in primary breast cancer of female Libyan patients with variable stage and grade and different treatment regimes. PATIENTS AND METHODS: Histological samples from 104 patients of breast carcinoma were retrospectively studied by computerized nuclear DNA cytometry. Isolated nuclei from paraffin sections were stained with Feulgen stain and DNA was measured using a computer-assisted image analysis cytometry system. In each case, 200 nuclei were measured and the DNA histograms, S phase fraction (SPF) and number of cells above 5c and 9c were determined. We applied different approaches in the analysis of DNA to compare the DNA histograms with different clinicopathological features and survival. RESULTS: The mean of DNA ploidy mode for all tumors was 3.43; 82.7% of tumors were aneuploid and 17.3% were diploid. The median SPF was 3.5% for DNA diploid and 13.5% for DNA aneuploid tumors. DNA aneuploid tumors and high SPF were associated with advanced stage, distant metastasis, high histological grade and lymph node involvement. The SPF was also associated with large tumor size and with younger patients (<50 years). In the overall population (median follow-up 51 months), patients with aneuploid DNA histograms and high SPF values had shorter survival times than those with diploid DNA histograms and low SPF values (p = 0.001, p < 0.0001, respectively). Also, short survival was associated with a multiploid DNA histogram and with DNA aneuploid cells ≥5 cells (p < 0.0001, p = 0.001, respectively). In a Cox multivariate analysis, DNA ploidy (p = 0.010), age (p = 0.038) and clinical stage (p = 0.001) were independent predictors of overall survival, and DNA ploidy (p = 0.018) and clinical stage (p = 0.001) also proved to be independent predictors of disease-specific survival. The SPF cutoff point of 11% might be applied to separate patients into good and poor prognosis groups. CONCLUSIONS: DNA image cytometry with careful analysis of the histograms may provide valuable prognostic information in Libyan breast cancer, with potential clinical implications in patient management, particularly in predicting the patients at high risk for metastasis and recurrence who should be considered as candidates for combined adjuvant therapy.

Estrogen receptor, progesterone receptor, and nuclear size features in female breast cancer in Libya: correlation with clinical features and survival.

BACKGROUND: The features of Libyan patients with breast cancer have not been fully investigated. The aim of this study was to evaluate the expression patterns of estrogen (ER) and progesterone receptor (PR), as well as nuclear morphometric features, in patients with breast cancer, and to correlate them with clinicopathological features and prognosis. PATIENTS AND METHODS: Data for a total of 62 female Libyan patients with breast cancer, diagnosed between 2000 and 2006, were retrospectively studied. Their clinical and pathological data were collected and analysed. Immunohistochemical evaluation of ER and PR expression was also performed. Further more nuclear morphometry was carried out. RESULTS: Of the 62 patients, disease in 10 was of the lobular type, 43 had invasive ductal and 9 had other carcinoma types; 47 out of 62 had lymph node involvement. Positive hormonal receptor expression was more common among those with lymph node-negative than lymph node-positive tumours. ER- and PR-positive patients appeared to have a better survival than ER- and PR-negative patients. The most significant difference, with respect to survival, was found between those bearing tumors with completely negative hormonal staining (J score 0) and those with positive staining (J score 1, 2 and 3). Larger nuclear size was associated with lymph node involvement and high-grade tumours (p<0.01 and p<0.0001, respectively), with shorter survival, larger tumour size and higher stage. CONCLUSION: The cut-off points for defining the groups with good or worse prognosis might be set, between score 0 and 1 (corresponding to 1% or fewer positive cells). Patients with ER- and PR-positive cancer had better overall survival than patients with hormonal receptor-negative cancer. In our hospital setting, ER and PR expressions and mean nuclear area (MNA) in breast carcinoma may be prognostically useful markers in guiding future treatment in prospective studies.

Loss of E-cadherin expression predicts disease recurrence and shorter survival in colorectal carcinoma.

The traditional staging system is currently inadequate for identifying those patients with colorectal carcinoma (CRC) who carry a high risk for poor outcome. In this study, the expression of E-cadherin was evaluated in CRC to determine its correlation with clinico-pathological variables, and association with disease outcome in patients with long-term follow-up. The present series consisted of tissue samples obtained from 230 patients with stage I, II, III, or IV CRC treated during 1981-1990 at Turku University Hospital. Archival paraffin-embedded samples were used to build up tissue microarray blocks, and E-cadherin expression was assessed by immunohistochemistry using an automated staining system. Different grading systems were tested for expression of E-cadherin. Fifty-nine percent of all tumors were positive for E-Cadherin. There was no significant correlation between E-cadherin expression and gender (p < 0.83), localization (p < 0.45), tumor invasion (p < 0.32), or histologic grade (p < 0.41). However, loss of E-cadherin expression was significantly associated with older age (p < 0.03) and lymph node involvement (p < 0.02), and with borderline significance with advanced stage (p < 0.09) and tumor metastasis (p < 0.09). In univariate (Kaplan-Meier) survival analysis, positive E-cadherin significantly (p = 0.009) predicted longer disease-free survival (DFS), and the same was true with disease-specific survival (DSS) as well (p = 0.007). In multivariate (Cox) survival analysis, E-cadherin retained its significance as independent predictor of DFS (HR = 1.56; 95% CI 1.01-2.42, p = 0.043), but not DSS. A sub-group analysis revealed that E-cadherin expression also predicts DFS (p < 0.01) and DSS (p < 0.04) in stage II CRC. Our results implicate the usefulness of E-cadherin expression in predicting disease recurrence and long-term survival in CRC.

Apoptotic activity in Libyan breast cancer.

BACKGROUND: We evaluated the relationship of the apoptotic activity index (AI) and the standardized mitotic-apoptotic ratio (SMI/AI) with clinicopathological features and prognosis in Libyan female breast cancer (BC) patients. We then compared our results with corresponding results in Finnish and Nigerian female BC patients. METHODS: Histological samples of breast carcinoma from 130 patients were retrospectively studied: an estimation of the apoptotic activity per square millimeter (expressed as apoptotic activity index (AI)), and standardized mitotic-apoptotic ratio (SMI/AI) was made, and the results compared with the clinicopathological features and the patient's survival. RESULTS: There was a statistically significant correlation between the AI and most of the clinicopathological features; the strongest association was observed for clinical stage lymph node (LN) status (P = 0.005). There were also correlations between AI and histological grade (P = 0.035), large tumor size (P = 0.011) and the clinical stage (P = 0.009). There were, however, prominent AI differences between Libyan, Nigerian and Finnish populations. The mean values of AI and SMI/AI in Libyan BC patients were 12.8 apoptotic figures per square millimeter and 2.8, respectively. The Libyan AI is slightly higher than in Nigeria, but much higher than in Finland. The differences between countries are seen throughout the samples as well as being present in certain subgroups. The survival analysis indicated that short survival time was associated with high apoptotic indices values and so can identify aggressive tumors and provide significant prognostic support. The cutoff (4 and 18 apoptosis/mm2) of AI might be applied as a quantitative criterion for Libyan BC to separate the patients into good, moderate and bad prognosis groups. CONCLUSIONS: The results indicated that the differences in AI among the three countries may be due to the known variation in the distribution of genetic markers in these populations. Improvement in health care and introduction of screening programs, however, could be very helpful in the Libyan population.

Prognostic value of mitotic counts in breast cancer of Saudi Arabian patients.

BACKGROUND: Quantitative methods in combination with other objective prognostic criteria can improve the evaluation of a cancer patient's prognosis, and possibly predict response to therapy. One of the important prognostic and predictive markers is the mitotic count, which has proven valuable in many aspects. In this study, the prognostic value of the mitotic count was assessed in breast cancer (BC) patients in Saudi Arabia. PATIENTS AND METHODS: The study comprised a series of 87 patients diagnosed and treated for breast cancer at the Departments of Surgery and Oncology, King Abdul-Aziz University Hospital, between 2000 and 2008. Mitotic counts were carried out using a standard laboratory microscope (objective, × 40; field diameter, 420 µm). The number of mitotic figures in 10 consecutive high-power fields (hpf) from the most cellular area of the sample gave the mitotic activity index (MAI, mitotic figures/10 hpf). The standardized mitotic index (SMI) recorded the mitotic count as the number of mitotic figures by area of the neoplastic tissue in the microscopic field, thus the number of mitoses in 10 consecutive fields was corrected for the volume fraction and field size (mitotic figures/mm²). RESULTS: The means of MAI and SMI of the tumors in the entire series of 87 patients were 15 mitotic figures/10 hpf (range 4-45) and 4 mitotic figures/mm² (range 1-9), respectively. The mitotic counts were higher in advanced stages than in early cancer (p < 0.04). The mitotic counts were significantly larger in patients with high-grade tumor (p < 0.004) and in cases with tumor metastasis (p < 0.004). The mitotic counts were also significantly larger in the recurrent cases than in non-recurrent ones (p < 0.02). CONCLUSION: The quantitatively measurable mitotic counts of cancer cell nuclei are of significant prognostic value in invasive ductal carcinoma of the breast in Saudi Arabia and the mean cut-off values of MAI and SMI can be applied as objective (quantitative) criteria to distinguish breast cancer patients into groups with favorable and less favorable prognosis.

Breast cancer patients in Libya: Comparison with European and central African patients.

The present study evaluated the incidence of breast cancer in Libya and described the clinicopathological and demographic features. These features were then compared with corresponding data from patients from sub-Saharan Africa (Nigeria) and Europe (Finland). The study consisted of 234 patients with breast carcinoma, admitted to the African Oncology Institute in Sabratha, Libya, during the years 2002-2006. The pathological features were collected from pathology reports, patient histories from hospital files and the Sabratha Cancer Registry. The demographic differences between the Libyan, Nigerian and Finnish populations were prominent. The mean age of breast cancer patients in Libya was 46 years which was almost identical to that of Nigeria, but much lower than that of Finland. The Libyan breast cancer incidence was evaluated as 18.8 per 100,000 female individuals. This incidence was markedly higher in Finland, but was also high in Nigeria. Libyan and Nigerian breast cancer is predominantly of premenopausal type and exhibits unfavorable characteristics such as high histological grade and stage, large tumor size and frequent lymph node metastases. However, the histological types and histopathological risk features show similar importance regarding survival as European breast cancer cases. Survival in Libya ranks between the rates of survival in Nigeria (lowest) and Finland (highest). In conclusion, in Libya and other African countries, premenopausal breast cancer is more common than postmenopausal breast cancer. However, the opposite is true for Europe. Population differences may be involved, as suggested by the known variation, in the distribution of genetic markers in these populations. Different types of environmental impacts, however, cannot be excluded.

Nuclear morphometry in prognostication of breast cancer in Saudi Arabian patients: comparison with European and African breast cancer.

BACKGROUND: The role of nuclear morphometry as a prognostic factor in breast cancer is well documented. The aim of this study was to evaluate this role in breast cancer in Saudi patients and to compare it with the experience in some African and European studies. PATIENTS AND METHODS: Primary tumors from 135 patients were analyzed using an image overlay drawing system (Prodit Morphometry Program), for the following nuclear features: area, perimeter, diameter, and roundness. RESULTS: The mean nuclear area (NA) was 93 microm(2) (range 45-168 microm(2)). The values of NA were higher in lymph node-positive patients than lymph node-negative patients and in advanced stages than early cancer. NA was significantly larger in patients with high grade tumor (p<0.0001) and in cases with tumor invasion (p<0.01). NA also was significantly larger in recurrent cases (103 microm(2)) than in non-recurrent ones (91 microm(2)). In univariate (Kaplan-Meier) analysis, NA was a significant predictor of disease-free survival (DFS) (log rank p<0.01), but not disease-specific survival (DSS). In multivariate (Cox) survival analysis, NA lost its significance as an independent predictor; response to treatment (p=0.0001) and tumor grade (p=0.030) being the only predictors of DFS. In a similar analysis for DSS, recurrence (p=0.040) and stage (p=0.003) were the only independent predictors. CONCLUSION: Nuclear morphometric profiles are helpful in identifying aggressive tumor phenotype (i.e. cases at risk for recurrence). The cut-off (93 mum(2)) of NA might be applied as quantitative criterion for Saudi female breast cancer to separate patients into good and poor prognosis groups. Mean NA of Saudi patients was markedly higher than the reported mean NA in the other studies and these differences might be due to technical variations or genetic bases.

Genomic analysis of the 55 kDa subunit of DNA polymerase epsilon in human intracranial neoplasms.

BACKGROUND: Defects of some DNA polymerases have shown cancer associations, but there are only limited data on DNA polymerase (Pol) epsilon. MATERIALS AND METHODS: We examined 26 human brain neoplasm DNA samples and 8 control blood samples (from Poland) for possible mutations in the entire coding region of the 55 kDa small subunit of human DNA Pol epsilon gene using polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) analysis, and sequence analysis of DNA. RESULTS: One single base intronic transition in intron 14 was found. The AATT deletion previously found in some breast and colorectal tumors was not found in samples from brain neoplasms or controls, but it was found in 1/100 normal blood samples from South-West Finland. CONCLUSION: We found no evidence that potential mutations in the 55 kDa subunit of DNA Pol epsilon are a contributing factor in the development of the tested cases of human intracranial tumors.

Image DNA cytometry in FNABs of Libyan breast disease.

BACKGROUND: The sensitivity for identification of malignant cells in conventional fine-needle aspiration biopsy (FNAB) investigation is about 80%. This percentage is dependent on the number of examined cells, type of breast cancer, and experience of the examiner. The aim of our study was to estimate the supporting value of image DNA cytometry of FNAB of the breast, and do so by using different sampling methods. MATERIALS AND METHODS: This retrospective study was based on 41 cases with an available histological diagnosis: 18 benign lesions and 23 malignant tumours were examined. The smears were submitted to image DNA analysis in a three-step protocol: (i) smears stained with HE method were destained and (ii) then restained with Feulgen staining for DNA and (iii) finally analysed using image cytometry. RESULTS: All non-malignant cases had diploid histogram. However, a few of them had one or two cells of >5c category. Most histologically malignant cases were aneuploid. Only three invasive ductal carcinomas showed diploid histograms. All samples with aneuploid histograms were malignant. CONCLUSION: The results confirm earlier published data in the Finnish population and indicate that image DNA cytometric analysis of nuclear content is a useful marker for identification of malignant cells in FNAB, especially after free cell sampling. The method can be used to increase the cytological sensitivity and specificity in doubtful breast lesions.

MMP-9 (gelatinase B) expression is associated with disease-free survival and disease-specific survival in colorectal cancer patients.

Extracellular matrix degradation is required for invasion and metastasis formation in colorectal carcinoma (CRC), therefore, we have examined matrix metalloproteinases MMP-9 expression in tumors from patients with CRC. The study comprises of 360 patients who underwent bowel resection for stage II, III, IV tumors. Paraffin-embedded CRC tissue samples were used for immunohistochemical staining. Negative MMP-9 expression levels correlated with longer survival time as evaluated by disease-free survival and disease-specific survival (p =.023, p =.006). In multivariate survival (Cox) analysis, MMP9 was a significant independent predictor of DFS (p =.014), but not of DSS, which was independently predicted by disease recurrence, stage and localization. The detection of MMP-9 expression may be valuable in finding patients who are at high risk of developing disease recurrence.

Prognostic significance of matrix metalloproteinase-9 (MMP-9) in stage II colorectal carcinoma.

BACKGROUND: Approximately 30% of all colorectal cancer patients are diagnosed with stage II disease. Adjuvant therapy is not widely recommended. However, it is well-established that a subgroup of patients with stage II is at high risk for recurrence within their life time and should be considered for adjuvant chemotherapy. The present work was designed to assess the value of matrix metalloproteinase-9 (MMP-9) as a predictor of disease outcome in a series of 202 stage II colorectal cancer (CRC) patients with long-term follow-up. METHODS: The present study comprises a series of 202 patients who underwent bowel resection for stage II CRC at Turku University Hospital. Archival paraffin-embedded CRC tissue samples were used to prepare tissue microarray blocks for immunohistochemical staining with MMP-9 antibody. RESULTS: Forty-eight percent of all CRC samples were positive for MMP-9. There was no significant correlation between MMP-9 expression and age, depth of invasion, and lymph node status. However, MMP-9 expression was significantly related to histological grade (p = 0.03) and location of the tumor (p = 0.01), therefore, being lower in high-grade tumors and most intense in carcinomas of the descending colon and rectum. Tumors with high MMP-9 expression showed a higher recurrence rate than tumors with low expression (p = 0.02). MMP-9 negative tumors had a more favorable disease-free survival (DFS) than those expressing MMP-9 (p = 0.03). The same was true with disease-specific survival (DSS; p = 0.02) as well, high expression of MMP-9 being associated with shorter survival rates. In multivariate (Cox) survival analysis, MMP-9 expression proved to be an independent predictor of DFS, but not DSS, which was predicted by age and sex only. CONCLUSION: Quantification of MMP-9 expression seems to provide valuable prognostic information in stage II CRC, particularly, in selecting the patients at high risk for recurrent disease who might benefit from adjuvant therapy.

Analysis of cyclins A, B1, D1 and E in breast cancer in relation to tumour grade and other prognostic factors.

BACKGROUND: The cell cycle is promoted by activation of cyclin dependent kinases (Cdks), which are regulated positively by cyclins and negatively by Cdk inhibitors. Proliferation of carcinoma is associated with altered regulation of the cell cycle. Little is known on the combined alterations of cyclins A, B1, D1 and E in breast cancer in relation to the tumour grade and other prognostic factors. FINDINGS: Immunohistochemical analysis of cyclins A, B1, D1 and E, estrogen receptor, progesterone receptor, Ki-67, Her-2/neu and CK5/6 was performed on 53 breast carcinomas. mRNA levels of the cyclins were analysed of 12 samples by RT-PCR. The expression of cyclins A, B1 and E correlated with each other, while cyclin D1 correlated with none of these cyclins. Cyclins A, B1 and E showed association with tumour grade, Her-2/neu and Ki-67. Cyclin E had a negative correlation with hormone receptors and a positive correlation with triple negative carcinomas. Cyclin D1 had a positive correlation with ER, PR and non-basal breast carcinomas. CONCLUSION: Cyclin A, B1 and E overexpression correlates to grade, Ki-67 and Her2/neu expression. Overexpression of cyclin D1 has a positive correlation with receptor status and non-basal carcinomas suggesting that cyclin D1 expression might be a marker of good prognosis. Combined analysis of cyclins indicates that cyclin A, B and E expression is similarly regulated, while other factors regulate cyclin D1 expression. The results suggest that the combined immunoreactivity of cyclins A, B1, D and E might be a useful prognostic factor in breast cancer.