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  • 1
    Keywords: CANCER ; CELLS ; tumor ; carcinoma ; CELL ; evaluation ; Germany ; incidence ; liver ; SAMPLE ; SAMPLES ; PATIENT ; MESSENGER-RNA ; primary ; CONTRAST ; LYMPH-NODES ; MR ; SEQUENCE ; BONE-MARROW ; STAGE ; PATTERNS ; ASSAY ; MUTATION ; COLORECTAL-CANCER ; chemotherapy ; MARKERS ; WILD-TYPE ; COLON-CANCER ; MUTATIONS ; ONCOGENE ; EPITHELIAL-CELLS ; CANCER-PATIENTS ; CARCINOMAS ; COLORECTAL CARCINOMAS ; MICROMETASTASES ; POLYMERASE-CHAIN-REACTION ; PROGNOSTIC-SIGNIFICANCE ; RT-PCR ; GREECE ; CANCER PATIENTS ; PERIPHERAL-BLOOD ; NODES ; COLORECTAL-CARCINOMA ; disseminated tumor cells,circulating epithelial cells,K-ras,cytokeratin 20,guanylylcyclase C ; GUANYLYL CYCLASE-C
    Abstract: The aim of this prospective study was to relate the incidences of cytokeratin 20 (CK20) and guanylylcyclase C (GCC) in lymph node, liver, and bone marrow specimens of 245 colorectal cancer (CRC) patients with the K-ras oncogene status of the corresponding primary tumor. Qualitative RT-PCR detection of CK20 and GCC mRNA was used as marker of circulating epithelial cells (CEC). Samples were considered positive for CEC only when both markers were detected concomitantly. For the detection of K-ras mutations, a PCR-RFLP assay was used. In the group with K-ras mutated primary carcinomas (n=92), CEC were detected in 62% of lymph node-, 43% of liver-, and 2% of bone marrow samples. No statistical significance was found when comparing these results with those from patients with K-ras wild-type carcinoma (59%, 46%, and 0%, respectively). In contrast to this combined evaluation, separate analysis of K-ras codons 12 (n=75, 82%) and 13 (n=17, 18%) revealed significantly differing CEC incidences. Lymph node specimens from corresponding K-ras codon 13 mutated carcinomas showed a significantly higher CEC incidence (82%) than the groups with codon 12 mutation (57%, p〈0.05) or K-ras wild-type sequence (59%, p〈0.05). Unlike these findings in lymph nodes, liver biopsies from corresponding carcinomas with K-ras codon 12 mutation or wild-type sequence were significantly more often positive for CEC (31% and 29%) than specimens from K-ras codon 13 mutated primary CRC (12%, p〈0.04, respectively). In conclusion, colorectal carcinomas with K-ras codon 12 mutation showed the same pattern of tumor cell dissemination as their K-ras wild-type counterparts. Since K-ras codon 12 mutations prevailed 4-fold over codon 13 mutations, combined analysis of the two codons showed the same result. However, sub-analysis of patients with K-ras codon 13 mutation revealed that the respective CEC incidence was significantly increased in lymph nodes, but decreased in liver biopsies
    Type of Publication: Journal article published
    PubMed ID: 15138598
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  • 2
    Keywords: CANCER ; CELLS ; tumor ; TUMOR-CELLS ; carcinoma ; CELL ; Germany ; CLASSIFICATION ; incidence ; liver ; NEW-YORK ; SAMPLE ; SAMPLES ; transcription ; TISSUE ; PATIENT ; primary ; TISSUES ; LYMPH-NODES ; MR ; bone marrow ; BONE-MARROW ; score ; STAGE ; ASSAY ; DIFFERENCE ; METASTASIS ; colorectal cancer ; COLORECTAL-CANCER ; chemotherapy ; MARKERS ; EPITHELIAL-CELLS ; CANCER-PATIENTS ; CARCINOEMBRYONIC ANTIGEN ; CARCINOMAS ; COLORECTAL CARCINOMAS ; CYTOKERATIN-20 ; DISSEMINATED TUMOR-CELLS ; dissemitiated tumor cells (DTC),hematogenic and lymphatic spread,RT-PCR detection of epithelial mark ; INVOLVEMENT ; MICROMETASTASES ; PERIPHERAL VENOUS-BLOOD ; POLYMERASE-CHAIN-REACTION ; PROGNOSTIC FACTORS ; PROGNOSTIC-SIGNIFICANCE ; RT-PCR
    Abstract: The aim of our prospective study was to detect circulating epithelial cells (CEC) indicating the presence of disseminated tumor cells (DTC) in tissues affected by lymphatic and hematogenic colorectal cancer metastasis. DTC were tracked in lymph node, liver or bone marrow samples of 245 colorectal cancer patients using 2 independent RT-PCR assays for cytokeratin 20 (CK20) and guanylylcyclase C (GCC) that demonstrated a sensitivity of 1 colorectal cancer cell in 10(6) nucleated hematopoietic cells. CK20 mRNA was detected in 79% of lymph nodes, 35% of both liver lobes and 11% of bone marrow samples. GCC mRNA was found in 68% of lymph nodes, 60% of both liver lobes and 6% of bone marrow specimens. Both markers were recorded in 63% of lymph nodes, 45% of at least 1 liver lobe and 1% of bone marrow samples. There was no significant difference when comparing lymph node samples tested positive for both markers in patients with (N1/2; 65%) and without (N0; 56%) nodal involvement. The same was true when comparing the percentages of patients with and without clinically overt distant metastasis who were positive for both markers in at least 1 liver lobe (62% vs. 41%) or in bone marrow (41% vs. 0%). A score denoting the cumulative sum of tests indicating presence of CK20 and GCC mRNA in the liver was significantly related with UICC classification (p = 0.039). However, addition of lymph node results to this score decreased the correlation. The high incidence of clinically inconspicuous lymph node and liver samples tested positive for both markers emphasizes the function of these organs as primary filters for epithelial cells possibly shed from colorectal carcinomas. The potential prognostic significance of these findings warrants verification, especially regarding the importance of CEC or DTC resident in the liver of collorectal cancer patients. (C) 2003 Wiley-Liss
    Type of Publication: Journal article published
    PubMed ID: 14520701
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  • 3
    Keywords: CANCER ; CELLS ; GROWTH ; carcinoma ; COMBINATION ; Germany ; liver ; SITE ; RNA ; PATIENT ; DNA ; MARKER ; BIOLOGY ; ASSAY ; MUTATION ; colorectal cancer ; COLORECTAL-CANCER ; metastases ; chemotherapy ; MARKERS ; RESECTION ; COLON-CANCER ; MUTATIONS ; CANCER PATIENTS ; K-RAS ; K-ras mutation ; RE ; KRAS MUTATIONS ; CYTOKERATIN 20 ; CODON ; circulating epithelial cells ; guanylylcyclase C ; K-RAS MUTATIONS ; KRAS ; liver metastasis
    Abstract: Aims. The aim of this study was to assess the incidence and lobar distribution of three surrogate tumour cell markers in biopsies from both liver lobes. Patients and methods. This study comprised 189 patients for whom DNA and/or RNA was available from both liver lobes and who showed at Least one positive marker in one liver lobe. Detection of cytokeratin 20 (CK20) and guanylylcyclase C (GCC) was performed by nested reverse transcription-PCR. For detection of K-ras mutations in codons 12 and 13, a PCR-restriction-fragment-length-polymorphism assay was used. Results. The incidence of all, markers and their combinations was higher in the smaller left lobe than in the larger right lobe (CK20: 62 vs 38%; GCC: 52 vs 48%; K-ras: 61 vs 39%; CK20+GCC: 61 vs 39%; CK20+GCC and/or K-ras: 61 vs 39%). The marker incidence in the two liver lobes was independent from the Location of the respective primary colorectal. carcinoma. Conclusions. The markers CK20, GCC, and K-ras indicating cells shed from the primary CRC were detected more often individually and in combination in biopsies from the smaller left lobe than from the Larger right lobe. The site of the primary tumour did not influence the marker incidence in both Liver lobes. (C) 2004 Elsevier Ltd. All rights reserved
    Type of Publication: Journal article published
    PubMed ID: 15642424
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  • 4
    Keywords: CANCER ; CELLS ; Germany ; liver ; PATIENT ; MESSENGER-RNA ; tumour ; LYMPH-NODES ; MR ; BONE-MARROW ; COLORECTAL-CANCER ; chemotherapy ; POLYMERASE-CHAIN-REACTION ; RT-PCR ; CANCER PATIENTS ; K-RAS ; RE ; GUANYLYL CYCLASE-C ; disseminated tumour cells ; RAS ONCOGENE MUTATIONS ; liver metastasis ; detection methods ; disseminated tumour cells (DTC) ; FRAGMENT-LENGTH-POLYMORPHISM
    Type of Publication: Journal article published
    PubMed ID: 16126360
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  • 5
    Keywords: CELLS ; GROWTH ; tumor ; CELL ; COMBINATION ; evaluation ; Germany ; MODEL ; THERAPY ; TOXICITY ; tumor growth ; HEPATOCELLULAR-CARCINOMA ; liver ; NEW-YORK ; TISSUE ; TUMORS ; RAT ; BODY-WEIGHT ; RATS ; MR ; treatment ; 5-FLUOROURACIL ; BREAST-CANCER ; FORM ; EFFICACY ; chemotherapy ; arteries ; chemoembolization,locoregional therapy,liver tumor,ethibloc ; COLORECTAL LIVER METASTASES ; HEPATOMA ; HEPATOMA-CELLS ; LONG-TERM SURVIVAL ; PROGNOSTIC-FACTORS ; RADIOFREQUENCY ABLATION ; RESECTION ; THERAPEUTIC EFFICACY ; tumor size
    Abstract: The aim of this study was to investigate experimentally whether there is a superior effect of the combination of hepatic artery chemo-embolization with portal vein infusion over either of the two treatment modalities alone. Novikoff hepatoma cells transplanted under the liver capsule of Sprague Dawley rats were used as a model. Tumor growth was assessed at 7 and 21 days after tumor inoculation. The prolamine solution Ethibloc was employed for embolization, and 5-fluorouracil was used as a chemotherapeutic agent for both infusion and chemo-embolization. All arterial treatment modalities were administered in a super-selective manner. There was no intolerable toxicity after dosages of 55 to 125 mg 5-fluorouracil/kg body weight. With regard to therapeutic efficacy the results show that embolization is an effective therapeutic means for inducing tumor necrosis in selected liver areas. As a consequence, the ranking of all treatment modalities was based on the combined evaluation of tumor size and extent of tumor necrosis. According to this evaluation, hepatic artery chemo-embolization was superior to the respective type of infusion (P〈0.01). In addition, the combination of both modalities in the form of hepatic artery chemo-embolization and portal vein infusion was effective in destroying more than 97% of vital tumor tissue (P〈0.01). These results suggest the need for a comparative clinical study
    Type of Publication: Journal article published
    PubMed ID: 14513367
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  • 6
    Keywords: CANCER ; CELLS ; SURVIVAL ; tumor ; TUMOR-CELLS ; carcinoma ; CELL ; Germany ; liver ; GENE ; SAMPLE ; SAMPLES ; TISSUE ; PATIENT ; DNA ; primary ; MR ; SEQUENCE ; polymorphism ; STAGE ; IDENTIFICATION ; ASSAY ; MUTATION ; METASTASIS ; chemotherapy ; MUTATIONS ; PARAMETERS ; CANCER-PATIENTS ; CARCINOMAS ; COLORECTAL CARCINOMAS ; PROGNOSTIC-SIGNIFICANCE ; CANCER-RESEARCH ; LIVER METASTASES ; SEQUENCE-ANALYSIS ; M1 ; COLORECTAL-CARCINOMA ; CLINICAL-SIGNIFICANCE ; LYMPH-NODE MICROMETASTASES
    Abstract: The aim of this study was to identify K-ras mutations as marker for isolated tumor cells in liver, lymph node, and bone marrow specimens of colorectal cancer patients. To detect these, a PCR-RFLP assay was used with a sensitivity exceeding that of routine histopathology by at least 1 order of magnitude. In addition, the ratio of mutated versus wildtype alleles was determined by an internal standard. Of 199 patients, 74 (37.5%) were found to bear a K-ras-positive tumor. Of these, 60 (81%) were mutated in codon 12 and 14 (19%) in codon 13 (P 〈 0.001). In addition, 14 organs were found K-ras positive, 13 of which were from 12 patients with a K-ras-positive tumor (16%) and 1 from a patient with a K-ras-negative tumor (0.8%). Eight patients exhibited liver involvement and 6 showed lymph node involvement. Remarkably, no bone marrow specimen was found K-ras positive (P 〈 0.017 versus liver involvement). Sequence analysis of tumor DNA revealed that GGT (Gly) was replaced by GAT (Asp; 35%), GTT (Val; 32%), AGT (Ser; 13%), GCT (Ala; 10%), TGT (Cys; 8%), and CGT (Arg; 2%) for codon 12, and by GAC (Asp) as the only type of mutation for codon 13. In colorectal carcinomas the ratio of K-ras mutated versus wild-type alleles ranged over 4 orders of magnitude (10(0)-10(-4), median: 10(-2)) and was correlated with both, residual tumor load (R1/2; P = 0.028) and distant metastasis (M1; P = 0.057). These results show that detection of K-ras mutated alleles by PCR-RFLP in patients with colorectal carcinoma may aid in the identification of isolated tumor cells. High ratios of K-ras alleles were correlated with certain negative prognostic parameters (R,M). In accord with its function as a primary filter for colorectal carcinoma cells, the liver was more often contaminated with K-ras-positive cells than bone marrow
    Type of Publication: Journal article published
    PubMed ID: 14760087
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  • 7
    Keywords: APOPTOSIS ; CANCER ; SYSTEM ; GENES ; PROTEIN ; DIFFERENTIATION ; BINDING ; BREAST-CANCER ; PROSTATE-CANCER ; chemotherapy ; COLON-CANCER ; CELL-GROWTH ; TUMOR-SUPPRESSOR ; I IGF-I ; IGFBP-3 ; VITAMIN-D ; siRNA silencing ; ISLAND METHYLATOR PHENOTYPE ; IGFBPs ; metastasis formation ; multi-functional genes ; tumor cell re-isolation
    Abstract: Over the last few decades, a great deal of attention has been directed to the IGF system for its vital role in regulating cell and tissue survival, growth and differentiation. The insulin-like growth factor binding proteins (IGFBPs), a main constituent of this system, have been implicated in the tumorigenesis of colorectal cancer (CRC). In this study, we intended to shed more light on two essential members; IGFBP3 as representative for the six main IGFBPs and IGFBP7 to represent their related proteins (IGFBP-rps). Our experiments on silencing IGFBP3 or IGFBP7 in the two human CRC cell lines SW480, Caco2, and in the rat CRC cell line CC531 show reduced proliferation, colony formation and for IGFBP3, also reduced migration. The expression of both genes in 68 human CRC samples was higher in UICC stages II and III than in stages I and IV. Additionally, IGFBP3 was negatively correlated with age (p = 0.05) and positively related to IGFBP7 expression (p = 0.0001). Further, in a liver metastasis experiment, the expression of both genes was drastically increased in response to early metastatic growth in vivo. Since these high levels returned gradually to normal thereafter, it could be assumed that the upregulation of IGFBPs is vital during the process of homing into the liver and early metastatic dissemination. Our results indicate that IGFBP3 and 7 cannot be simply considered as tumor suppressors but have additional properties, which become evident only during cancer progression and metastasis formation
    Type of Publication: Journal article published
    PubMed ID: 21525788
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  • 8
    Keywords: CELLS ; ENDOTHELIAL GROWTH-FACTOR ; METASTASIS ; HUMAN COLON-CANCER ; FAMILIAL ADENOMATOUS POLYPOSIS ; FACTOR-BETA ; TGF-BETA ; CYCLOOXYGENASE-2 ; OSTEOPONTIN ; PCR DATA
    Abstract: PURPOSE: Colorectal cancer (CRC) is driven by genetic alterations causing its progression. Besides accepted tumor suppressor- and onco- genes, a series of genes have been identified, which contribute to transformation into a more malignant stage. We investigated whether the expression level of such genes, alone or in combination, could add to predict the prognosis of CRC patients. METHODS: Tumor samples from 118 CRC patients were screened in a retrospective analysis by qRT-PCR for expression of the four tumor progression-associated genes osteopontin (Opn), transforming growth factor beta (Tgf-beta), matrix metalloproteinase-2 (Mmp-2) and cyclooxigenase-2 (Cox-2). The resulting qRT-PCR values were related to those of housekeeping genes. All patients were clustered for similar expression levels between the four genes with R statistical software using the package pvclust, which provides bootstrap agglomerative hierarchical clustering. Clusters with similar expression of the four genes were analyzed for correlation with UICC stages and survival time. RESULTS: Expression of the four genes varied considerably within the cohort of patients. Cluster analysis of patients revealed a subgroup (n = 33) who in comparison with the other patients showed tenfold higher expression levels of all four genes (p 〈 0.001, respectively). However, there was no correlation between patients expressing high or low levels of these four genes and known parameters of clinical prognosis (UICC stages, survival time). CONCLUSIONS: In conclusion, tenfold increased expression levels of Opn, Tgf-beta, Mmp-2 and Cox-2 in a subset of CRC patients did not predict for a clinical outcome that is different from that of the remaining patients.
    Type of Publication: Journal article published
    PubMed ID: 22614156
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  • 9
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    German Medical Science; Düsseldorf, Köln
    In:  122. Kongress der Deutschen Gesellschaft für Chirurgie; 20050405-20050408; München; DOC05dgch2949 /20050615/
    Publication Date: 2005-06-16
    Keywords: ddc: 610
    Language: German
    Type: conferenceObject
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  • 10
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    German Medical Science; Düsseldorf, Köln
    In:  122. Kongress der Deutschen Gesellschaft für Chirurgie; 20050405-20050408; München; DOC05dgch2959 /20050615/
    Publication Date: 2005-06-16
    Keywords: ddc: 610
    Language: German
    Type: conferenceObject
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