[Show abstract][Hide abstract] ABSTRACT: Background
Unlike metastatic colorectal cancer (CRC) there are to date few reports concerning the predictive value of molecular biomarkers on the clinical outcome in stage II/III CRC patients receiving adjuvant chemotherapy. Aim of this study was to assess the predictive value of proteins related with the EGFR- and VEGFR- signalling cascades in these patients.Methods
The patients' data examined in this study were from the collective of the 5-FU/FA versus 5-FU/FA/irinotecan phase III FOGT-4 trial. Tumor tissues were stained by immunohistochemistry for VEGF-C, VEGF-D, VEGFR-3, Hif-1 ¿, PTEN, AREG and EREG expression and evaluated by two independent, blinded investigators.Survival analyses were calculated for all patients receiving adjuvant chemotherapy in relation to expression of all makers above.ResultsPatients with negative AREG and EREG expression on their tumor had a significant longer DFS in comparison to AREG/EREG positive ones (p< 0.05). The benefit on DFS in AREG-/EREG- patients was even stronger in the group that received 5-FU/FA/irinotecan as adjuvant treatment (p=0.002). Patients with strong expression of PTEN profited more in terms of OS under adjuvant treatment containing irinotecan (p< 0.05). Regarding markers of the VEGFR- pathway we found no correlation of VEGF-C- and VEGFR-3 expression with clinical outcome. Patients with negative VEGF-D expression had a trend to live longer when treated with 5-FU/FA (p=0.106). Patients who were negative for Hif-1 ¿, were disease-free in more than 50% at the end of the study and showed significant longer DFS-rates than those positive for Hif-1 ¿ (p=0.007). This benefit was even stronger at the group treated with 5-FU/FA/irinotecan (p=0.026). Finally, AREG-/EREG-/PTEN+ patients showed a trend to live longer under combined treatment combination.Conclusions
The addition of irinotecan to adjuvant treatment with 5-FU/FA does not provide OS or DFS benefit in patients with stage II/III CRC. Nevertheless, AREG/EREG negative, PTEN positive and Hif-1 ¿ negative patients might profit significantly in terms of DFS from a treatment containing fluoropyrimidines and irinotecan. Our results suggest a predictive value of these biomarkers concerning adjuvant chemotherapy with 5-FU/FA +/¿ irinotecan in stage II/III colorectal cancer.
Full-text · Article · Oct 2014 · Journal of Experimental & Clinical Cancer Research
[Show abstract][Hide abstract] ABSTRACT: BACKGROUND: Our aim was to compare long-term results of adjuvant treatment of colon cancer (CC) and rectal cancer (RC). Adjuvant chemotherapy of CC improved overall survival (OS), whereas that of RC remained at the level achieved by 5-fluorouracil (5-FU). METHODS: We separately conducted 2 identically designed adjuvant trials in CC and RC. Patients were assigned to adjuvant chemotherapy with 5-FU alone, 5-FU + folinic acid (FA), or 5-FU + interferon-alfa. The first study enrolled patients with stage IIb/III CC, and the second study enrolled patients with stage II/III RC. All patients with RC received postoperative irradiation. RESULTS: Median follow-up for all patients with CC (n = 855) and RC (n = 796) was 4.9 years. The pattern and frequency of recurrence differed significantly, especially lung metastases, which occurred more frequently in RC (12.7%) than in CC (7.3%; P < .001). Seven-year OS rates for 5-FU, 5-FU + FA, and 5-FU + IFN-alfa were 54.1% (95% confidence interval [CI], 46.5-61.0), 66.8% (95% CI, 59.4-73.1), and 56.7% (95% CI, 49.3-63.4) in CC and 50.6% (95% CI, 43.0-57.7), 56.3% (95% CI, 49.4-62.7), and 54.8% (95% CI, 46.7-62.2) in RC, respectively. A subgroup analysis pointed to a reduced local recurrence (LR) rate and an increased OS by the addition of FA in stage II RC (n = 271) but not in stage III RC (n = 525). CONCLUSION: FA increased 7-year OS by 12.7 percentage points in CC but was not effective in RC. Based on these results and the pattern of metastases, our results suggest that the chemosensitivity of CC and RC may be different. Strategies different from those used in CC may be successful to decrease the frequency of distant metastases in RC in the future.
No preview · Article · Oct 2012 · Clinical Colorectal Cancer
[Show abstract][Hide abstract] ABSTRACT: Background/aims:
Thymidylate synthase (TS) is an enzyme for DNA-synthesis and the target for 5-fluorouracil whereas cyclin-D1 plays a critical role in the progression of cells through the G1 phase of the cell-cycle. There is evidence that expression of these markers may predict the outcome of patients with colorectal cancers. The aim of this study was to examine the prognostic value of TS and cyclin D1 protein expression in patients with node negative colorectal cancers.
TS and cyclin D1 protein expression from 140 patients with UICC stage I and II colorectal cancer was analyzed by immunhistochemistry in paraffin-embedded primary tumour specimens.
The 1-, 5- and 10-year overall-survival rates were 96%, 86% and 71%, respectively. Tumour stage and recurrence were associated with overall-survival. Low- and high TS immunoreactivity was present in 68 (48%) and 72 (52%) of cancers, respectively. Low- and high cyclin D1 immunoreactivity was present in 98 (70%) and 42 (30%) of the cancers, respectively. Patients (n=72) with high TS expressing tumours had a worse overall-survival than patients (n=68) with low TS expressing colorectal cancers (p=0.011). No difference in overall-survival was seen between patients with high and low cyclin D1 expressing cancers.
TS may be helpful as a prognostic marker in lymph node negative colorectal cancer.
No preview · Article · Sep 2012 · Hepato-gastroenterology
[Show abstract][Hide abstract] ABSTRACT: Gastric cancer is the fourth cancer in incidence worldwide, with a high disease-related death rate. The aim of this study was to evaluate the importance of clinical and pathological factors for the prognosis of gastric cancer patients.
We analysed data from 304 consecutive patients. Clinical and pathological factors and the surgical resection status were analysed by univariate analyses, followed by investigation of important factors using a proportional hazard regression analysis with backward elimination in order to identify important independent prognostic factors.
Univariate analysis revealed that age, pT, pN, M, UICC stage, grading, and resection status were significantly associated with survival. Multivariate analysis identified age, pT, pN, M, and resection status as independent prognostic factors.
Besides age and pathological parameters, radical R0 resection plays an essential role in the management of gastric cancer and should be aimed at, even if extended resection may be necessary.
No preview · Article · May 2012 · Anticancer research
[Show abstract][Hide abstract] ABSTRACT: Interleukin-4 (IL-4) and interleukin-13 (IL-13) are anti-inflammatory and immunomodulatory cytokines which can influence cancer-directed immunosurveillance. Nothing is presently known about expression of these cytokines and their receptors (IL-4R and IL-13R) in colorectal cancer. The aim of this study was to characterize their expression in primary colorectal cancer specimens and to evaluate possible functions for this disease.
Expression of IL-4, IL-13, IL-4R, and IL-13R protein was characterized by immunohistochemistry in 359 patients with Union for International Cancer Control stage I-III colorectal cancer and evaluated by uni- and multivariate analysis for their prognostic relevance.
All four proteins were expressed in colorectal cancer specimens. In the cancer cells, high IL-4, IL-13, IL-4R, and IL-13R immunoreactivity were present in 33 % (118/359), 50 % (181/359), 36 % (129/359), and 42 % (152/359), respectively. Patients with high expression of IL-4, IL-4R, and IL-13R had a lower frequency of lymph node metastases. Expression of IL-13 did not influence the frequency of lymph node metastases. However, high IL-13-immunoreactivity was associated with a better overall survival (p = 0.041). Expression of IL-4, IL-4R, or IL-13R did not influence survival. Multivariate analysis revealed that besides pT classification and tumor recurrence, IL-13 expression was an independent prognostic factor for overall survival.
Expression of IL-4, IL-4R, and IL-13R are involved in the process of local metastases in colorectal cancer, while IL-13 expression has an impact on survival. These interleukins and their receptors may become attractive targets for the treatment of colorectal cancer.
No preview · Article · Mar 2012 · International Journal of Colorectal Disease
[Show abstract][Hide abstract] ABSTRACT: High-level microsatellite instability (MSI-H) has been reported as a prognostic marker in colon cancer. We here analysed the prognostic significance of MSI and mutations of the Beta2-Microglobulin (B2M) gene, which occur in about 30% of MSI-H colon cancer, in the cohort of the prospective FOGT-4 (Forschungsruppe Onkologie Gastrointestinale Tumoren, FOGT) trial.
Microsatellite instability status was determined using standard protocols (NCI/ICG-HNPCC panel and CAT25) in 223 colon cancer lesions. Beta2-Microglobulin mutation status was evaluated by exon-wise sequencing in all MSI-H lesions.
Patients with MSI-H (n=34) colon cancer presented with a significantly lower risk of relapse after 12 months of follow-up compared with MSS (n=189) colon cancer patients (5 year time to relapse: MSI-H 0.82 vs MSS 0.66, P=0.03). No significant difference in overall survival was detected. Beta2-Microglobulin mutations were identified in 10 (29.4%) out of 34 MSI-H colon cancers and were associated with a complete absence of disease relapse or tumour-related death events (P=0.09).
The risk of late disease relapse was significantly lower in patients with MSI-H compared with MSS colon cancer. Moreover, B2M mutations may contribute to the favourable outcome of MSI-H colon cancer patients and should therefore be evaluated as a potential prognostic marker in future clinical trials.
Full-text · Article · Feb 2012 · British Journal of Cancer
[Show abstract][Hide abstract] ABSTRACT: Objective: Traumatic diaphragmatic injuries are rare, but potentially life-threatening due to herniation of abdominal organs into the pleural cavities. They can be easily overlooked on initial diagnostics and a high index of suspicion is required. The aim of this retrospective study was to analyze the clinical presentation, diagnostic methods and surgical management of patients with diaphragmatic rupture at our institution.
Methods: A retrospective study was performed to analyze our experience with patients suffering from traumatic diaphragmatic rupture. Charts were reviewed for sex, age, side-location, concomitant injuries, time-to-diagnosis, diagnostic methods, surgical approach and outcome.
Results: Fourteen patients (median age: 46 yrs, range 18–71, 9 male, 5 female) with diaphragmatic injuries (left side: 10, right side: 4) were treated between July 2003 and September 2011. Mechanism of injury was a penetrating trauma (14%), blunt trauma (50%) and others (36%). Associated abdominal injuries included spleen rupture (n=3), liver laceration (n=2), abdominal wall laceration (n=2) and gastric perforation (n=1). Computed tomography was the most sensitive diagnostic method. All patients underwent trans-abdominal repair of the diaphragmatic defect (direct suture: 10, prosthetic mesh insertion: 4). Associated abdominal procedures included splenectomy (n=3), liver packing (n=2), abdominal wall reconstruction (n=2) and partial gastric resection (n=1). Morbidity and hospital mortality rate were 36% and 0%, respectively. Median postoperative hospital stay was 17 days (range: 7–40 days).
Conclusion: Morbidity and mortality of diaphragmatic ruptures are mainly determined by associated injuries or complications of diaphragmatic herniation like incarceration of viscera or lung failure. Early diagnosis helps to prevent severe complications. Spiral CT-scan is the most reliable tool for acute diagnosis of diaphragmatic rupture and associated visceral lacerations. Laparotomy is an adequate surgical approach for diaphragmatic repair, especially in cases of associated abdominal organ injury.
[Show abstract][Hide abstract] ABSTRACT: Cells transfected with DHFR or TS gene specific siRNAs maintain similar proliferation rate compared to the negative control. (A and B) Western immunoblot analysis of protein expression levels of DHFR and TS by siRNAs against DHFR or TS. (C and D) The impacts of siRNAs against DHFR or TS on the cell proliferation. Non-specific siRNA (negative control) was used as the negative control.
[Show abstract][Hide abstract] ABSTRACT: Translational control mediated by non-coding microRNAs (miRNAs) plays a key role in the mechanism of cellular resistance to anti-cancer drug treatment. Dihydrofolate reductase (DHFR) and thymidylate synthase (TYMS, TS) are two of the most important targets for antifolate- and fluoropyrimidine-based chemotherapies in the past 50 years. In this study, we investigated the roles of miR-215 in the chemoresistance to DHFR inhibitor methotrexate (MTX) and TS inhibitor Tomudex (TDX).
The protein levels of both DHFR and TS were suppressed by miR-215 without the alteration of the target mRNA transcript levels. Interestingly, despite the down-regulation of DHFR and TS proteins, ectopic expression of miR-215 resulted in a decreased sensitivity to MTX and TDX. Paradoxically, gene-specific small-interfering RNAs (siRNAs) against DHFR or TS had the opposite effect, increasing sensitivity to MTX and TDX. Further studies revealed that over-expression of miR-215 inhibited cell proliferation and triggered cell cycle arrest at G2 phase, and that this effect was accompanied by a p53-dependent up-regulation of p21. The inhibitory effect on cell proliferation was more pronounced in cell lines containing wild-type p53, but was not seen in cells transfected with siRNAs against DHFR or TS. Moreover, denticleless protein homolog (DTL), a cell cycle-regulated nuclear and centrosome protein, was confirmed to be one of the critical targets of miR-215, and knock-down of DTL by siRNA resulted in enhanced G2-arrest, p53 and p21 induction, and reduced cell proliferation. Additionally, cells subjected to siRNA against DTL exhibited increased chemoresistance to MTX and TDX. Endogenous miR-215 was elevated about 3-fold in CD133+HI/CD44+HI colon cancer stem cells that exhibit slow proliferating rate and chemoresistance compared to control bulk CD133+/CD44+ colon cancer cells.
Taken together, our results indicate that miR-215, through the suppression of DTL expression, induces a decreased cell proliferation by causing G2-arrest, thereby leading to an increase in chemoresistance to MTX and TDX. The findings of this study suggest that miR-215 may play a significant role in the mechanism of tumor chemoresistance and it may have a unique potential as a novel biomarker candidate.
[Show abstract][Hide abstract] ABSTRACT: miR-215 expression is decreased in colorectal cancer compared to normal colorectal specimens by real time qRT-PCR analysis. Expression level of miR-215 was normalized by the internal control RNU6B in each sample. P = 0.0002, two-tailed paired Wilcoxon test.
[Show abstract][Hide abstract] ABSTRACT: Antagonizing miR-215 by LNA anti-miR reverses the impact of miR-215 on the cell cycle. (A) Knock-down miR-215 decreased G2 phase and increased S phase of the cell cycle in HCT 116 (wt-p53) cells, no such effects were found in HCT 116 (null-p53) cells. HCT 116 (wt-p53) cells and HCT 116 (null-p53) cells were transfected with 100 nM miR-215 for 24 h. Cell cycle analysis was performed after transfected with 100 nM LNA-miR215 for 48 h. (B) In parallel, LNA-miR215 prevented the induction of p53 and p21 expression in HCT 116 (wt-p53) cells analyzed by Western immunoblot analysis. LNA-control was the negative control. This experiment was repeated two separate times, and similar results were obtained. The representative flow cytometry pattern was shown.
[Show abstract][Hide abstract] ABSTRACT: Knock-down of endogenous miR-215 enhances the cell proliferation and chemosensitivity to MTX. (A) HCT 116 (wt-p53) cells were transfected with 100 nM of scramble-miR locked nucleic acid (LNA-control) or LNA anti-miR215 oligonucleotide (LNA-miR215) by Lipofectamine 2000, cell proliferation analysis was performed as described in Additional file 2. (B) HCT 116 (wt-p53) cells were transfected with LNA-control or LNA-miR215 and treated with MTX for 72 h, viable cells were accessed by WST-1 assays. (C) Proteins were extracted at 48 h after transfection with LNA-miR215 and subjected to Western immunoblot analysis to detect DHFR and TS, LNA-control was used as the negative control.
[Show abstract][Hide abstract] ABSTRACT: miR-215 has no effect on the cytotoxicity of cisplatin and doxorubicin. HCT 116 (wt-p53) cells transfected with miR-215 mimics, non-specific miRNA, or non-targeting siRNA and siRNA against DTL were incubated with cispaltin (0.625-10 μM) or doxorubicin (25-500 nM) for 72 h and cell viability was measured by WST-1 at 450 and 630 nm respectively.
[Show abstract][Hide abstract] ABSTRACT: The expression of miR-215 in human colon cancer stem cells is elevated. (A) FACS analysis was performed to sort colon cancer stem cells using CD133 and CD44 as the markers. CD133+HI/CD44+HI cells were considered as the colon cancer stem cells. CD133+/CD44+ and CD133NEG/CD44NEG were considered as the colon cancer cells. (B) Expression of miR-215 in human colon cancer stem cells was analyzed by real-time qRT-PCR. The value of miR-215 in the CD133+/CD44+ colon cancer cells was set at 1, the relative amount in CD133+HI/CD44+HI colon cancer stem cells and CD133NEG/CD44NEG colon cancer cells was showed as the fold induction. (C) The expression of DHFR and TS proteins was decreased in CD133+HI/CD44+HI colon cancer stem cells compared to control cell population analyzed by Western immunoblot analysis.
[Show abstract][Hide abstract] ABSTRACT: Real time qRT-PCR analysis of DHFR mRNA (A) or TS mRNA levels (B) in HCT-116 (wt-p53) cells transfected with miR-215 or siRNAs specific for DHFR or TS. Oligofectamine alone (vehicle control) and non-specific siRNA (negative control) were negative controls, the value of DHFR mRNA or TS mRNA in the negative control was set at 1, the relative amount in siRNAs against DHFR and TS or miR-215 transfected cells was indicated as fold induction. *P < 0.05, compared to the negative control, Student's t test (two-tailed). Each condition was repeated 3 times and error bars represent standard deviations.