Prognostic relevance of autophagy-related markers LC3, p62/sequestosome 1, Beclin-1 and ULK1 in colorectal cancer patients with respect to KRAS mutational status.

BACKGROUND: Autophagy is a cellular pathway that regulates transportation of cytoplasmic macromolecules and organelles to lysosomes for degradation. Autophagy is involved in both tumorigenesis and tumour suppression. Here we investigated the potential prognostic value of the autophagy-related proteins Beclin-1, p62, LC3 and uncoordinated (UNC) 51-like kinase 1 (ULK1) in a cohort of colorectal cancer (CRC) specimens. METHODS: In this study, we analysed the immunoexpression of the autophagy-related proteins p62, LC3, Beclin-1 and ULK1 in 127 CRC patients with known KRAS mutational status and detailed clinical follow-up. RESULTS: Survival analysis of p62 staining showed a significant correlation of cytoplasmic (not nuclear) p62 expression with a favourable tumour-specific overall survival (OS). The prognostic power of cytoplasmic p62 was found in the KRAS-mutated subgroup but was lost in the KRAS wildtype subgroup. Survival analysis of Beclin-1 staining did not show an association with OS in the complete cohort. LC3 overexpression demonstrated a slight, though not significant, association with decreased OS. Upon stratifying cases by KRAS mutational status, nuclear (not cytoplasmic) Beclin-1 staining was associated with a significantly decreased OS in the KRAS-mutated subgroup but not in the KRAS wildtype CRCs. In addition, LC3 overexpression was significantly associated with decreased OS in the KRAS-mutated CRC subgroup. ULK1 expression was not correlated to survival. CONCLUSIONS: Immunohistochemical analyses of LC3, p62 and Beclin-1 may constitute promising novel prognostic markers in CRC, especially in KRAS-mutated CRCs. This strategy might help in identifying high-risk patients who would benefit from autophagy-related anticancer drugs.

Quantification of ischemia-reperfusion injury to the small intestine using a macroscopic score.

INTRODUCTION: Classical methods do not allow to rapidly quantify the heterogeneously distributed ischemia-reperfusion injury along the small intestine. We therefore established a suitable macroscopic score. METHODS: A rat model based on superior mesenteric artery occlusion (45 or 90 min ischemia; 0, 120, or 180 min reperfusion) was used on 42 rats. RESULTS: Subsequent to ischemia-reperfusion or pure ischemia, macroscopic phenotypes of the small intestine were defined and differentiated based on the severity of existing petechiae and hemorrhages (macroscopic types 0-1-2-3). Analysis of the interobserver variability verified the reliability of this macroscopic differentiation. Macroscopic types were directly correlated with their histological alterations (Chiu score). On the basis of the sample hemoglobin content, a nonlinear macroscopic score (0-1-3-9) was derived from the linearly related macroscopic types. Mean macroscopic scores after 45 min ischemia/180 min reperfusion were lower than those after 90 min ischemia/120 min reperfusion. Nevertheless, all scores correlated well with their respective hemoglobin contents (R(2) = 0.87). Heterogeneous patterns of macroscopic scores that depended on the ischemic period and largely differed between individual animals were found to be distributed along the small intestine with the highest injury score at the last half of the jejunum. The iron chelator deferoxamine mesylate clearly decreased the overall tissue hemoglobin content and macroscopic score of the small intestine but local protection was restricted to the proximal and middle part of the jejunum. CONCLUSION: The local as well as overall ischemia-reperfusion injury in the rat small intestine can be rapidly and reliably assessed macroscopically.

Differential expression of microRNA 181b and microRNA 21 in hyperplastic polyps and sessile serrated adenomas of the colon.

This study was designed to analyse the potential diagnostic value of miR-181b and miR-21 for discriminating hyperplastic polyps (HP) from sessile serrated adenomas (SSA) without cytologic dysplasia. Using real-time polymerase chain reaction expression levels of miR-181b and miR-21 in 18 HPs, 19 SSAs without cytologic dysplasia and 20 normal colonic mucosal specimens were examined. In addition, 20 colorectal cancers specimen were analysed for miR-181b expression. Data were normalised to RNU48 as an internal control. A differential expression of miR-181b and miR-21 was found in HPs, SSAs, and normal colonic mucosa with highest expression levels in SSAs. Levels of miR-181b but not miR-21 differed in HPs and normal mucosa. SSAs exhibited both significantly higher miR-181b levels (up to 2.01-fold; P < 0.001) and miR-21 levels (up to 1.82-fold; P = 0.011) than HPs. In contrast to HPs, SSAs are characterised by high levels of miR-181b and miR-21 expression. However, due to the overlap of values, miR-181b and miR-21 evaluation did not allow discrimination of the two lesions in every case.

Activation of the ERK and AKT signalling pathway predicts poor prognosis in hepatocellular carcinoma and ERK activation in cancer tissue is associated with hepatitis C virus infection.

BACKGROUND/AIMS: The aim of the study was to determine the prognostic relevance of AKT and extracellular regulated kinases (ERK1/2), which are implied in the regulation of cell proliferation and apoptosis, in hepatocellular carcinoma (HCC). METHODS: This study comprised a series of 208 patients incorporating HCCs treated either by surgical resection (n = 109) or liver transplantation (n = 99). Immunohistochemically demonstrated phospho-ERK1/2 (pERK1/2) and phospho-AKT (pAKT) was correlated with a series of clinico-pathologically relevant parameters (EGFR, Cyclin-D1, HCV/HBV infection, liver cirrhosis, chronic alcohol abuse), proliferative activity, and apoptosis. RESULTS: Activation of ERK1/2 correlated statistically with the presence of HCV infection. pERK1/2 (P < 0.001) and pAKT (P = 0.052) expression showed a significant correlation with a decreased overall survival (OS). In multivariate Cox regression analysis pERK1/2 was identified as an independent prognostic parameter in HCC (P = 0.026). CONCLUSIONS: Activation of ERK1/2 in HCC cancer indicates aggressive tumour behaviour and constitutes an independent prognostic factor. Furthermore our data confirm that HCV infection activates the ERK pathway and thereby might contribute to HCC carcinogenesis. Immunohistochemical determination of pERK1/2 status can thus be proposed as a promising candidate for the identification of high risk patients who may benefit from new anticancer drugs targeting the ERK-pathway.

Laser-tonsillotomy for treatment of obstructive tonsillar hyperplasia in early childhood: a retrospective review.

INTRODUCTION: Tonsillar hyperplasia leading to dyspnea, dysphagia and other symptoms of obstruction represents a common problem especially in young children where tonsillectomy should be avoided in order to preserve the immunological function of the tonsils. Aim of the study was to assess carbon-dioxide-laser-tonsillotomy as a considered alternative procedure to reduce the tonsillar volume in these children. METHODS: Between 1993 and 2004, 109 children with tonsillar hyperplasia without former episodes of tonsillitis received laser-tonsillotomy mostly (n=98) combined with adenoidectomy. The protruding part of the tonsil was reduced by a CO2-laser. Seventy-five children were available for follow-up with a standard questionnaire. Five patients required a subsequent tonsillectomy due to a recurrence of tonsillar hyperplasia. Histological investigations were performed. Twenty-two children were reevaluated by clinical examination. RESULTS: Most of the patients were relieved from obstructive symptoms. There was no occurrence of postoperative hemorrhage or peritonsillar abscesses. The histological investigations on the specimens from later performed tonsillectomy (n=5) showed no evidence of inflammation or scar formations, but open and deep crypts. The clinical examination did not reveal any signs of chronic infections. CONCLUSION: In this retrospective study tonsillotomy with CO2-laser in early childhood leads to a long-term elimination of obstructive symptoms due to tonsillar hyperplasia with minimal discomfort for the patient while preserving normally functioning immunocompetent tonsillar tissue. Further prospective studies are planned.