[Show abstract][Hide abstract] ABSTRACT: Colorectal cancer (CRC) accounts for high mortality. So far, there is lack of markers capable of predicting which patients are at risk of aggressive course of the disease. Protein phosphatase-2A (PP2A) inhibitor proteins have recently gained interest as markers of more aggressive disease in certain cancers. Here, we report the role of PP2A inhibitor PME-1 in CRC. PME-1 expression was assessed from a rectal cancer patient cohort by immunohistochemistry, and correlations were performed for various clinicopathological variables and patient survival. Rectal cancer patients with higher cytoplasmic PME-1 protein expression (above median) had less recurrences (P = 0.003, n = 195) and better disease-free survival (DFS) than the patients with low cytoplasmic PME-1 protein expression (below median). Analysis of PPME-1 mRNA expression from TCGA dataset of colon and rectal adenocarcinoma (COADREAD) patient cohort confirmed high PPME1 expression as an independent protective factor predicting favorable overall survival (OS) (P = 0.005, n = 396) compared to patients with low PPME1 expression. CRC cell lines were used to study the effect of PME-1 knockdown by siRNA on cell survival. Contrary to other cancer types, PME-1 inhibition in CRC cell lines did not reduce the viability of cells or the expression of active phosphorylated AKT and ERK proteins. In conclusion, PME-1 expression predicts for a favorable outcome of CRC patients. The unexpected role of PME-1 in CRC in contrast with the oncogenic role of PP2A inhibitor proteins in other malignancies warrants further studies of cancer-specific function for each of these proteins.
Cancer Medicine 09/2015; DOI:10.1002/cam4.541 · 2.50 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Aim: The present study is undertaken to evaluate E-cadherin expression in a series of Libyan colorectal cancer cases to get an insight in its potential prognostic value in colorectal cancer in Libyan patients. Materials and Methods: A series of 81 Libyan patients with colorectal carcinoma were retrospectively studied. All carcinomas were selected from the archives of the Department of pathology, Benghazi University, derived from the period from January 2007 to December 2011. All tumors were classified using the histopathological criteria of the World Health Organization (WHO) classification, and staging was made according to the criteria of tumor-node-metastasis (TNM) classification of the International Union against Cancer. Results and Discussion: Immuno Histo chemical (IHC) analysis was done using the automatic system (BenchMark XT, Ventana Medical System, Inc. Tucson, Arizona, USA). This fully automated processing of code-labeled slides included baking of the slides, solvent free deparaffinization, antigen retrieval in a cell conditioning buffer CCI (Mild: 36 minutes conditioning, and standard: Two staining indexes were calculated: the membrane index (MI) and cytoplasmic index (CI). These indices were calculated with both the intensity of staining and the fraction of positively-stained cells taken into account using the following formula: I= 0 x f0 + 1 x f1 + 2 x f2 + 3 x f3 Our data showed that loss of E-cadherin expression is more frequently detected in older age group and in colorectal cancer patients with lymph node involvement; 75% of tumors with lymph node involvement showed negative expression of E-cadherin. One of the most important finding of the current study is the association of E-cadherin expression with the disease outcome. Conclusion: These data suggest that the loss of the E-cadherin function could be associated with invasiveness, lymph node metastasis and distant metastasis resulting in poor prognosis.
[Show abstract][Hide abstract] ABSTRACT: Objective: The aim of the current study is to cast further light on the issues related to prognostication of renal cell carcinoma (RCC), assessing the expression of survivin in a subset of primary RCC and determine its relation to different clinicopathological features and disease free survival. Methods: The present series consisted of tissue samples obtained from 37 Libyan patients with stage I, II, III, or IV RCC. Survivin expression in these tumors was assessed by immunohistochemistry using an automated staining system. Different grading systems were tested for expression of survivin. Results: Expression of survivin was significantly associated with venous invasion (tumor thrombus) (p= 0.042), larger tumor size (p= 0.051), higher primary T classification (p= 0.013), advanced tumor stage (p= 0.033), and borderline association (p= 0.068) with tumor location. In univariate (Kaplan-Meier) survival analysis, survivin expression showed a borderline association (p= 0.089) with disease-free survival (DFS). However, there was no significant correlation between survivin expression and gender, age, histological grade, distance metastasis, lymph node involvement, perinephric fat and capsular invasion, status at end point and recurrence. Conclusion: Survivin expression in RCC may identify patients at risk of a more aggressive disease and a worse prognosis, further investigations, on a larger and more heterogeneous population, should be carried out to validate and extend our results.
European Journal of Cancer 10/2013; 1(4):184-194. DOI:10.5455/jihp.20130324114215 · 5.42 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: 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.
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).
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).
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.
Anticancer research 08/2013; 33(8):3137-43. · 1.83 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Little is known about the acute and subacute toxicity of Alhagi Graecorum (Camel Thorn) in mice. The aim of this study is to investigate the acute toxicity (LD50) and sub acute toxicity of camel thorn in mice. For determination of LD50, mice weighing 25-30 g were divided into nine groups each of 6 animals and received 130, 330, 660, 1300, 2600, 4200, 5000, 5900 and 6900 mg/kg orally of camel thorn water extract respectively. The sign and symptoms of toxicity and the number of died animals in each group were registered and the LD50 was calculated.Another group of Male albino mice weighed 25-30 g were divided into control and different treated group each of 6 mice and placed in the metabolism cages that allowed daily measurement of food and water consumption. The control received normal saline whereas the other groups received 130, 660, 1300 mg/kg (i.p, daily) of camel thorn extract (CTE)respectively, for two weeks.. The animals were weighed daily for any sign of reduction or gain of body weight. The food and water consumption were daily calculated. At the end of this experiment, the mice were killed and blood was collected and used for the determination of glucose and the serum creatinine,urea, aspartate, aminotransferase, (AST) and alanine aminotransferase (ALT) levels.The liver of the animals were removed,fixed in 10%formaline for histopathological investigation. The LD50 was equal to 5400mg/kg.Results of sub acute toxicity studies revealed that no significant weight reduction were observed in treated groups as compared to control, however the food consumption was significantly increased in the mice received 660mg/kg of CTE . Also the water consumption was significantly decreased in the animal receiving 1300mg/kg of CTE.
International Journal of Applied Biology and Pharmaceutical Technology 07/2013; 4(3):202-209. · 0.99 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: 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.
[Show abstract][Hide abstract] ABSTRACT: 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.
[Show abstract][Hide abstract] ABSTRACT: The frequency of over-expression of human epidermal growth factor receptor-2 (HER-2) in bladder cancer is one of the highest among all human malignancies. This over-expression is thought to play a role in aberrant proliferation of cancer cells. Studies on HER-2 expression in bladder carcinoma have shown heterogeneous results.
The aim of the study was to evaluate the status of HER-2 protein expression in patients with invasive carcinomas of the urinary bladder as related to tumor grade and stage.
Archival samples from 39 patients (6 women, 33 males) with urinary bladder cancer were analyzed for HER-2 over-expression, using immunohistochemistry with the HercepTest.
HER-2 over-expression was observed in 23/39 tumors (59%) and was more frequent in high-grade than in low-grade carcinomas, but the difference was not statistically significant. A significant correlation was established between HER-2 over-expression and tumor stage (p=0.011). HER-2 expression was more frequent in transitional cell carcinomas (TCC) and adenocarcinomas (AC) as compared with squamous cell carcinoma (SCC). Patients' age and sex were not related to HER-2 over-expression.
Over-expression of HER-2 was frequent in carcinomas of the urinary bladder. Knowing the HER-2 status would be helpful in formulating a rational treatment strategy for patients with urinary bladder cancer.
Libyan Journal of Medicine 03/2012; 7(1). DOI:10.3402/ljm.v7i0.14694 · 1.06 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Among the patients with bladder cancer, a group is still at risk of disease recurrence, progression, and death from their cancer after curative treatment. Angiogenesis is a crucial pathogenic mechanism for this type of urothelial carcinoma and is a potential therapeutic target.
To quantify tumor angiogenesis in bladder cancer and determine whether it correlates with tumor stage and grade.
A series of 42 archival samples from carcinomas of the urinary bladder were graded, staged, and analyzed for microvessel density (MVD) by a double immunohistochemical technique using Factor VIII (FVIII) and CD31 antibodies. The correlation between MVD and histopathological grade and tumor stage was evaluated.
FVIII and CD31 immunoreactivity was observed in 100% of cases and more intensely with CD31. Significantly higher MVD was determined in invasive tumors than in superficial tumors (p<0.05). MVD increased with tumor grade and stage (p<0.05); MVD was not affected by age or sex of the patients.
These data demonstrate that MVD in bladder carcinoma correlates with the tumor grade and stage. Quantification of tumor angiogenesis may allow selection of the type of treatment for bladder cancer patients.
Libyan Journal of Medicine 03/2011; 6(1). DOI:10.3402/ljm.v6i0.6016 · 1.06 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The expression of EGFR and p53 has not been adequately studied as a prognostic tool in urinary bladder tumors. We analyzed 74 bladder cancer samples from Egypt for EGFR and p53 expression using immunohistochemistry. The tumors were of different histological types, grades and clinical stages, and with established lymph node status. Almost 61% of the tumors showed positive membranous EGFR expression and 74.3% had positive nuclear staining of p53. Analysis of correlation of the IHC staining with clinical variables showed a significant correlation only between EGFR expression and histological type (p=0.002, ANOVA), in that the expression was higher in squamous cell carcinomas than in other histological types. There were no significant correlations between p53 or EGFR with the other clinicopathological variables, including age, sex, staging, grading, and lymph node status. Further studies are needed to determine if EGFR and p53 might be used as prognostic tools in bladder cancer.
Libyan Journal of Medicine 12/2009; 4(4):143-5. DOI:10.4176/090625 · 1.06 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Approximately 30% of all colorectal cancer (CRC) 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 are at high risk for recurrence within their lifetime and should be considered for adjuvant chemotherapy. The present work was designed to study the prognostic value of nuclear DNA content in stage II CRC of patients with long-term followup.
Isolated nuclei from 50 microm-thick paraffin sections of tissue samples from 253 patients with stage II CRC, who had undergone bowel resection at Turku University Central Hospital were cytocentrifuged on slides, stained with Feulgen staining, and DNA was measured using a computer-assisted image analysis cytometry system. Different approaches were applied in analysis of DNA histograms.
DNA content did not show any relation with age (p < 0.96), sex (p < 0.35), tumor invasion (p < 0.77), or grade (p < 0.31). Aneuploid DNA content was significantly more frequent in the cancer of the left colon and rectum than the right colon (p = 0.02). S-phase fraction analysis revealed that a higher proportion (62%) of the older patients (>65 years) had high proliferation rates than did the younger patients (p < 0.05). Patients with narrow range histograms had a better disease-free survival (DFS) (narrow range: 70%, wide range: 60% at 10 years). Tumors with >9c nuclei were associated with significantly better DFS and disease-specific survival (DSS) as compared with the patients who did not have >9c nuclei in their tumor samples (p < 0.003 and p < 0.0001, respectively). Multivariate survival (Cox) model showed that only classification of the basic pattern of the histogram [odds ratio OR) = 29.14; 95% confidence interval (CI) 2.350-361.57] (p = 0.009) and recurrence (OR = 165.35; 95% CI 48.42-564.7) (p = 0.0001) proved to be independent predictors of clinical outcome.
Our results seem to suggest it truly is possible, by using DNA cytometry, to find groups with different prognosis among stage II cases. Those with a high recurrence rate should be considered for adjuvant chemotherapy.
Anticancer research 02/2009; 29(1):99-106. · 1.83 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Ezrin is a membrane-cytoskeleton anchor, which, in experimental models, regulates tumor cell invasion and metastatic ability. We carried out immunohistochemical analysis of ezrin in 74 advanced colorectal cancer patients and correlated it to clinicopathologic variables and disease outcome. In contrast to the predominantly membraneous immunoreactivity of normal colorectal epithelium, ezrin expression in the colorectal cells was typically cytoplasmic. Altogether, 16.2% (12/74) of the tumors showed negative/weak ezrin staining, 35.1% (26/74) had moderate staining, and 48.6% (36/74) had intense staining. The expression was more intense in colon than in rectal carcinomas (P = .003). Increased ezrin expression was associated with adverse outcome, that is, shorter disease-specific survival; 48.3 months and 36.6 months for negative-weak versus intense expression (P = .041) as well as shorter survival with metastases at 36 months (P = .030); the metastases(36) rates in ezrin(neg/weak), ezrin(moderate), ezrin(intense) are 58.3%, 25.0%, and 18.4%, respectively. In univariate survival analysis, dichotomized (negative/weak versus moderate/strong) ezrin expression significantly predicted both the 5-year disease specific survival (P = .035) and 5-year metastases (P = .018) but lost this predictive power in multivariate (Cox) analysis. High ezrin expression was also related to high E-cadherin (cytoplasmic) expression, DNA aneuploidy, and high thymidylate synthase expression (P = .046, P = .042, P = .046, respectively). These results suggest that ezrin may play a role in colorectal cancer progression and that ezrin expression might provide clinically valuable information in predicting the biological behavior of colorectal cancer.
Human pathology 09/2008; 39(12):1737-43. DOI:10.1016/j.humpath.2008.04.020 · 2.77 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: To investigate the changing pattern of alpha-catenin expression and its relationship to clinical and pathological features of colorectal cancer (CRC) patients.
Archival tumor samples were analyzed using immunohistochemistry (IHC) for alpha-catenin in 91 patients with advanced CRC.
The values of alpha-catenin membrane index (MI) and cytoplasmic index (CI) were significantly related to the depth of tumor invasion (P = 0.027, P = 0.020, respectively), high indices being associated with increased depth of the primary tumor invasion (T3 and T4). Similarly, patients with high alpha-catenin expression had a significantly increased risk of lymph node metastasis (32/39 vs 37/52 for MI and 37/45 vs 32/46 for CI) (P = 0.001, P = 0.0001, respectively, for LNN status). An altered expression (i.e., cytoplasmic pattern) was also related (P = 0.047) to the response to chemotherapy; patients with low CI were more responsive (CR: 7/46) than patients with high CI values (CR: 0/45). There was a marginal effect on survival in patients time with metastases (SWM) (P = 0.087); patients with low CI showing slightly longer SWM, but no such effect on disease free survival (DFS) or disease specific survival (DSS). As to co-expression with another member of the adhesion complex (beta-catenin), high alpha-catenin/beta-catenin MI index was of marginal significance in predicting longer DSS (P = 0.063, log-rank).
The results implicate that high alpha-catenin expression is intimately involved in the key regulatory mechanisms leading to invasive phenotype, lymph node metastases, and progressive disease in CRC.
World Journal of Gastroenterology 09/2008; 14(31):4903-8. · 2.37 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: To investigate the changing pattern of beta-catenin expression and its prognostic value in advanced colorectal cancer (CRC).
Archival tumor samples were analyzed for beta-catenin using immunohistochemistry (IHC) in 95 patients with advanced CRC.
Membranous beta-catenin expression was found in the normal colorectal epithelium. Almost 100% of CRC cases showed membranous and cytoplasmic expression, and 55 (58%) cases showed nuclear expression. In univariate (Kaplan-Meier) survival analysis, only the nuclear index (NI) was a significant predictor of disease free survival (DFS) (P = 0.023; n = 35), with a NI above the median associated with longer DFS (34.2 mo) than those with a NI below the median (15.5 mo) (P = 0.045, ANOVA). The other indices were not significant predictors of DFS, and none of the three tested indices (for membranous, cytoplasmic, or nuclear expression) predicted disease-specific survival (DSS). However, when dichotomized as positive or negative nuclear expression, the former was a significant predictor of more favorable DFS (P = 0.041) and DSS (P = 0.046).
Nuclear beta-catenin expression provides additional information in predicting patient outcome in advanced CRC.
World Journal of Gastroenterology 07/2008; 14(24):3866-71. · 2.37 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: To study the dynamic events leading to impaired cell-cell adhesion upon transition to the invasive phenotype of colorectal cancer (CRC), we examined three distinct beta-catenin expression patterns (membranous, cytoplasmic, and nuclear) in the paired samples of the primary tumours (P) and their metastatic lesions (M). beta-catenin expression was detected by immunohistochemistry (IHC) in 33 pairs of the primary CRC and their metastases. In a pair-wise (P-M) comparison, the membranous index (MI) was significantly different between P and M (p=0.036, Wilcoxon Signed-Ranks test), while cytoplasmic index (CI) and nuclear index (NI) values did not significantly deviate between P and M. MI in primary tumours was inversely related to the patient's age (p=0.04) and tumour grade (p=0.03), while patients with low MI in M had a high rate of metastasis at diagnosis (p=0.06). CI in P was lower in patients with LN involvement (p=0.02) and in advanced tumour stage (p=0.002). Tumours of the ascending colon had the highest CI in their M (p=0.04). Interestingly, high MI of the M lesions was a significant predictor of favourable overall survival (OS) in univariate (Kaplan-Meier) survival analysis (p=0.035). In conclusion, significant aberrations in beta-catenin expression probably take place in CRC cells during the development of metastatic phenotype, but a change from membrane expression to cytoplamic and/or nuclear expression is not a prerequisite for metastasis in all cases.
[Show abstract][Hide abstract] ABSTRACT: AIM: To investigate the changing pattern of β-catenin expression and its prognostic value in advanced colorectal cancer (CRC). METHODS: Archival tumor samples were analyzed for β-catenin using immunohistochemistry (IHC) in 95 patients with advanced CRC. RESULTS: Membranous β-catenin expression was found in the normal colorectal epithelium. Almost 100% of CRC cases showed membranous and cytoplasmic expression, and 55 (58%) cases showed nuclear expression. In univariate (Kaplan-Meier) survival analysis, only the nuclear index (NI) was a significant predictor of disease-free survival (DFS) ( P = 0.023; n = 35), with a NI above the median associated with longer DFS (34.2 mo) than those with a NI below the median (15.5 mo) (P = 0.045, ANOVA). The other indices were not significant predictors of DFS, and none of the three tested indices (for membranous, cytoplasmic, or nuclear expression) predicted disease- specific survival (DSS). However, when dichotomized as positive or negative nuclear expression, the former was a significant predictor of more favorable DFS ( P = 0.041) and DSS (P = 0.046). CONCLUSION: Nuclear β-catenin expression provides additional information in predicting patient outcome in advanced CRC.
World Journal of Gastroenterology 01/2008; 14(24). DOI:10.3748/wjg.14.3866 · 2.37 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Thymidylate synthase (TS) is the rate-limiting enzyme in the synthesis of pyrimidine nucleotides and as such a critical target for fluoropyrimidines, which are widely used in the treatment of colorectal cancer (CRC). The purpose of this study was to investigate TS expression in the primary tumours (PTs) and their metastases (M) in advanced CRC.
TS expression was determined immunohistochemically in paraffin-embedded biopsies of PT-M pairs in 39 CRC patients, as related to the clinical data.
There was no difference in the mean TS index of PTs compared with that of M, 1.25 and 1.14, respectively (p=0.12). TS expression of PTs was above the mean more often than that of M (61.5% and 41.0%, respectively, p=0.035). High TS expression in PTs was significantly related to high expression in M (the Fisher exact test, p=0.001). Using the absolute index values, TS expression in PT and M was significantly correlated (Pearson R=0.501, p=0.001). In 29/39 (74.3%) pairs, PT and M had concordant expression levels (Cohen's kappa 0.508, 95% CI 0.260-0.756, p=0.001; intraclass correlation coefficient (ICC) = 0.679, 95% CI 0.358-0.836, p=0.0001). No significant association was found between TS expression and any of the clinicopathological variables, disease outcome (DFS, DSS) or its response to treatment in univariate or multivariate analysis.
Albeit usually higher, TS expression in PT was closely correlated with TS expression in M. This suggests that measurement of TS in primary CRC accurately predicts TS expression in subsequent metastases, which may help in selecting those patients most likely to respond to 5-FU-based regimens.