Publications

BACKGROUND

Autophagy is characterized by the sequestration of cytoplasm and organelles into multimembrane vesicles and subsequent degradation by the cell's lysosomal system. It is linked to many physiological functions in human cells including stress response, protein degradation, organelle turnover, caspase-independent cell death and tumor suppression. Malignant transformation is frequently associated with deregulation of autophagy and several tumor suppressors can modulate autophagic processes. The tumor suppressor p53 can induce autophagy after metabolic or genotoxic stress through transcriptionally-dependent and -independent mechanisms. In this study we expand on the former mechanism by functionally characterizing a p53 family target gene, ISG20L1 under conditions of genotoxic stress.

RESULTS

We identified a p53 target gene, ISG20L1, and show that transcription of the gene can be regulated by all three p53 family members (p53, p63, and p73). We generated an antibody to ISG20L1 and found that it localizes to the nucleolar and perinucleolar regions of the nucleus and its protein levels increase in a p53- and p73-dependent manner after various forms of genotoxic stress. When ectopically expressed in epithelial cancer-derived cell lines, ISG20L1 expression decreased clonogenic survival without a concomitant elevation in apoptosis and this effect was partially rescued in cells that were ATG5 deficient. Knockdown of ISG20L1 did not alter 5-FU induced apoptosis as assessed by PARP and caspase-3 cleavage, sub-G1 content, and DNA laddering. Thus, we investigated the role of ISG20L1 in autophagy, a process commonly associated with type II cell death, and found that ISG20L1 knockdown decreased levels of autophagic vacuoles and LC3-II after genotoxic stress as assessed by electron microscopy, biochemical, and immunohistochemical measurements of LC3-II.

CONCLUSIONS

Our identification of ISG20L1 as a p53 family target and discovery that modulation of this target can regulate autophagic processes further strengthens the connection between p53 signaling and autophagy. Given the keen interest in targeting autophagy as an anticancer therapeutic approach in tumor cells that are defective in apoptosis, investigation of genes and signaling pathways involved in cell death associated with autophagy is critical.

OBJECTIVE

To compare the accuracy of radiography and computed tomography (CT) in predicting implant position in relation to the vertebral canal in the cervical and thoracolumbar vertebral column.

STUDY DESIGN

In vitro imaging and anatomic study.

ANIMALS

Medium-sized canine cadaver vertebral columns (n=12).

METHODS

Steinmann pins were inserted into cervical and thoracolumbar vertebrae based on established landmarks but without predetermination of vertebral canal violation. Radiographs and CT images were obtained and evaluated by 6 individuals. A random subset of pins was evaluated for ability to distinguish left from right pins on radiographs. The ability to correctly identify vertebral canal penetration for all pins was assessed both on radiographs and CT. Spines were then anatomically prepared and visual examination of pin penetration into the canal served as the gold standard.

RESULTS

Left/right accuracy was 93.1%. Overall sensitivity of radiographs and CT to detect vertebral canal penetration by an implant were significantly different and estimated as 50.7% and 93.4%, respectively (P<.0001). Sensitivity was significantly higher for complete versus partial penetration and for radiologists compared with nonradiologists for both imaging modalities. Overall specificity of radiographs and CT to detect vertebral canal penetration was 82.9% and 86.4%, respectively (P=.049).

CONCLUSIONS

CT was superior to radiographic assessment and is the recommended imaging modality to assess penetration into the vertebral canal.

CLINICAL RELEVANCE

CT is significantly more accurate in identifying vertebral canal violation by Steinmann pins and should be performed postoperatively to assess implant position.