Expression of the EP300, TP53 and BAX genes in colorectal cancer: Correlations with clinicopathological parameters and survival.

E1A binding protein P300 (EP300), tumor protein P53 (TP53) and BCL2 associated X, apoptosis regulator (BAX) genes encode proteins which cooperate to regulate important cellular processes. The present study aimed to determine the expression levels of EP300, TP53 and BAX in colorectal cancer (CRC) and to investigate their prognostic value and association with the progression of CRC. Tumor and matched unchanged colorectal tissues were collected from 121 CRC patients. Quantitative polymerase chain reaction and immunohistochemistry were used to assess the mRNA and protein levels of the studied genes. Altered expression of the studied genes in CRC tissues was observed at both the mRNA and protein levels. The depth of invasion was associated with TP53 mRNA levels and was correlated negatively with BAX mRNA expression. Moreover, a relationship between tumor location and BAX mRNA content was noted. BAX immunoreactivity was correlated positively with the intensity of p300 immunostaining and was associated with lymph node involvement and tumor-node-metastasis (TNM) disease stage. Univariate regression analysis revealed that overexpression of p53 and BAX in CRC tissues was associated with poor patient outcome. In conclusion, dysregulation of the expression of the studied genes was found to contribute to CRC pathogenesis. The association between p300 and BAX levels suggests the existence of an interdependent regulatory mechanism of their expression. Moreover, BAX expression may be regulated alternatively, in a p53-independent manner, since the lack of correlations between expression of these factors was observed.

Altered expression of the PLAGL1 (ZAC1/LOT1) gene in colorectal cancer: Correlations to the clinicopathological parameters.

Pleomorphic adenoma gene-like 1 gene (PLAGL1) encodes a zinc-finger nuclear transcription factor which promotes apoptosis and cell cycle arrest. Loss or downregulation of its expression has been observed in various human neoplasms. This study compared PLAGL1 expression in colorectal cancer (CRC) tissue and colon mucosa of healthy subjects at the mRNA and protein levels, and estimated its prognostic value. The PLAGL1 mRNA levels were also determined in CRC cell lines. We collected paired tumor tissue and unchanged mucosa of the large intestine from 121 CRC patients as well as 72 colon biopsies of healthy subjects obtained during screening colonoscopy. PLAGL1 mRNA levels were determined by quantitative PCR, while PLAGL1 protein expression was estimated by western blotting and immunohistochemistry. PLAGL1 mRNA level in tumor tissue was ~2-fold lower than in samples of corresponding unchanged tissues and biopsies of healthy colon mucosa. Downregulated expression of PLAGL1 mRNA was also observed in all tested CRC cell lines. Although the average content of PLAGL1 protein did not differ significantly between tumor and unchanged tissues of CRC patients or colon mucosa of healthy individuals, the decreased PLAGL1 protein levels in tumor specimens correlated with lymph node involvement, the presence of metastases and higher TNM disease stage. The PLAGL1 expression level did not correlate significantly with patient overall survival; however, the hazard ratio for patients whose tumor tissues showed reduced PLAGL1 immunohistochemical staining was twice higher than in patients with increased PLAGL1 immunoreactivity. In conclusion, these results suggest that dysregulation of PLAGL1 expression may be involved to some extent in the progression of CRC, but the so far collected patient survival data do not confirm applicability of the PLAGL1 expression level as a prognostic factor in CRC.

PLAGL1 protein is differentially expressed in the nephron segments and collecting ducts in human kidney.

INTRODUCTION: PLAGL1 (pleiomorphic adenoma gene-like 1) is a C2H2-type zinc finger transcription factor associated with the regulation of cell growth and development. Although PLAGL1 expression in kidney was assessed by biochemical methods, the exact localization of the PLAGL1 protein in human kidney has not yet been described. MATERIAL AND METHODS: Macroscopically unchanged specimens of kidney tissue were collected from 39 patients undergoing nephrectomy due to renal cell carcinoma. H & E staining of paraffin sections was used to assess histology of the kidney whereas immunohistochemistry was used to localize PLAGL1 protein in kidney compartments. In addition, database sequences search for putative PLAGL1 binding sites among the kidney-related genes was performed. RESULTS: PLAGL1 staining intensity differed depending on the kidney compartment. Strong PLAGL1 immunoreactivity was found in thick ascending limbs of Henle's loop, distal tubules and collecting ducts, whereas PLAGL1 expression in proximal tubules and renal corpuscles (including podocytes) was moderate and weak, respectively. By the in sillico screening of promoter sequences for PLAGL1 specific DNA-binding sites GGG-GCCCC we designated 43 candidate genes for PLAGL1-regulated genes. Analysis of their functional annotations identified three significantly over-represented gene sets: inositol phosphate metabolic processes (GO), endocrine and other factor-regulated calcium reabsorption (KEGG) and calcium signaling pathways (KEGG). CONCLUSION: Differences in the renal expression of PLAGL1 suggest that this protein may be involved in the regulation of several cellular pathways both as transcriptional factor and coactivator/corepressor of other tran-scription factors reflecting its role in the cell type-specific control of gene expression.

[Microphthalmos and hypertelorism as diagnostic index in ultrasound diagnosis of Fraser syndrome]. / Maloocze i szerokie rozstawienie galek ocznych w obrazie usg jako wskaznik diagnostyczny obecnosci zespolu Frasera u plodu.

Fraser syndrome is a rare genetic syndrome with abnormalities of the head, lungs, kidneys, and limbs. A prenatal diagnosis of FS can be done in families with risk, using foetal ultrasonography. However, a wide qualitative and quantitative variability of possible abnormalities makes the diagnosis in utero notably questionable. We present the results of foetal ultrasonography in a tertigravida, had delivered two children with FS. Signs of foetal hypertelorism and microphthalmia, both traits typical for FS, were detected based on outer and inner orbital diameters and ocular diameters in 28 and 32 weeks of pregnancy. The clinical and pathological examinations after birth confirmed the diagnosis of FS. Our observation suggests that eye anomalies may prompt the diagnosis of FS even if characteristic lung and kidney abnormalities are absent. Therefore, we propose to regularly assess eye dimensions and distance, when performing any foetal ultrasonography in families with of FS.

Fetotoxic action of 2,4-dichlorophenoxyacetic acid (2,4-D). III. Morphological changes in rat kidneys.

The paper presents the findings of histological studies on the effect of sodium salt of 2,4-D acid on the changes within kidneys in newborn rats exposed to this herbicide in the prenatal and postnatal period. The experiment was performed on 60 Wistar rats of both sexes, up to 10 weeks of age. The animals were divided into two groups: I group (control)--18 rats fed on a standard diet and given tap water ad libitum, and group II (experimental)--42 rats, whose mothers received sodium salt of 2.4 dichlorophenoxyacetic acid in drinking water at a daily dose of c. 250 mg/kg for 2 months before fertilization and during pregnancy and lactation. The animals were killed after 24 hours, 4, 6 and 10 weeks of the experiment. The sections were taken from the kidneys, fixed in 4% formaldehyde and stained with hematoxylin and eosin. For acid and alkaline phosphatase examination, the kidney section were fixed in Backer's liquid and Gomori histoenzymatic reaction was performed. Histological examination of the first four experimental groups revealed changes in kidney tubules. Histologic changes were nonspecific and a variety of conditions. The presence vacuoles in cytoplasm and necrosis of tubular epithelial cells. Varying degrees of isometric vacuolization of proximal tubular epithelium, tubular microfocal calcification, tubular epithelial inclusion bodies and peritubular capillary congestion were observed. The observations suggest that chronic intoxication with 2,4-D acid leads to renal cell damage in kidneys more intensified in the fetal than in the postnatal period. Following herbicide withdrawal, the most pronounced changes observed in the fetus were found to regress.

Bcl-2 immunohistochemical detection in prostate cancer.

Prostate cancer (Pca) is the most prevalent cancer diagnosed and the second leading cause of cancer-related deaths among men in the Poland. The present study was designed to analyse the relationship between expression of Bcl-2 protein in prostate cancer (PCa) following radical prostatectomy to chosen anatomoclinical and morfological parameters of the tumours. Tumours from 28 patients were assessed by immunohistochemistry. Tissue sections were fixed in 10% buffered formaldehyde solution, embedded in paraffin and stained immunohistochemically with the anti-human Bcl-2 antibody (Dako/Clon124). The immunolocalization of Bcl-2 was performed using the Labelled Streptavidin Biotin (LSAB) method. No correlation was found between Bcl-2 protein expression and pT stage, lymph node metastases, Gleason score, seminal vesicles invasion, positive or negative resection margins as well as capsular penetration and preoperative PSA serum level.

Expression of Ki-67 and PCNA as proliferating markers in prostate cancer.

Prostate carcinoma (PCa) is the most common male cancer in industrialized societies and represents a serious public health problem. The aim of our study was the immunohistochemical evaluation of Ki-67 and proliferating cell nuclear antigen (PCNA) expression in prostate cancer (PCa) following radical prostatectomy and analysis of its relationship to chosen anatomo-clinical and morfological parameters of the tumours. Archival sections from 28 PCas were immunostained with monoclonal antibodies against Ki-67, and proliferating cell nuclear antigen (PCNA). Immunolocalization of Ki-67 and PCNA was performed using LSAB method. No statistically significant correlation was found between the expression of Ki-67, PCNA and preoperative PSA level, lymph node metastases, capsular penetration, seminal vesicle invasive and positive or negative surgical resection margins. However, a strong statistically significant correlation between Ki-67 positive and T stage was found. We also found relationship between Gleason score 7 or above and high expression of Ki-67 and PCNA in prostate cancers (p < 0.004, p < 0.02 respectively). These results suggest that PCNA and especially Ki-67 may be useful as tumour markers in prostate carcinoma, but further studies on larger group are required.