miRNA Clusters with Down-Regulated Expression in Human Colorectal Cancer and Their Regulation.

Regulation of microRNA (miRNA) expression has been extensively studied with respect to colorectal cancer (CRC), since CRC is one of the leading causes of cancer mortality worldwide. Transcriptional control of miRNAs creating clusters can be, to some extent, estimated from cluster position on a chromosome. Levels of miRNAs are also controlled by miRNAs "sponging" by long non-coding RNAs (ncRNAs). Both types of miRNA regulation strongly influence their function. We focused on clusters of miRNAs found to be down-regulated in CRC, containing miR-1, let-7, miR-15, miR-16, miR-99, miR-100, miR-125, miR-133, miR-143, miR-145, miR-192, miR-194, miR-195, miR-206, miR-215, miR-302, miR-367 and miR-497 and analysed their genome position, regulation and functions. Only evidence provided with the use of CRC in vivo and/or in vitro models was taken into consideration. Comprehensive research revealed that down-regulated miRNA clusters in CRC are mostly located in a gene intron and, in a majority of cases, miRNA clusters possess cluster-specific transcriptional regulation. For all selected clusters, regulation mediated by long ncRNA was experimentally demonstrated in CRC, at least in one cluster member. Oncostatic functions were predominantly linked with the reviewed miRNAs, and their high expression was usually associated with better survival. These findings implicate the potential of down-regulated clusters in CRC to become promising multi-targets for therapeutic manipulation.

Expression of miR-34a-5p is up-regulated in human colorectal cancer and correlates with survival and clock gene PER2 expression.

Colorectal cancer represents a leading cause of cancer death. MicroRNAs (miRNAs) are small non-coding RNA molecules that have been extensively studied in tumours, since changes in their levels can reveal patient prognosis. Cancer progression is also influenced by the circadian system whose functioning is based on the rhythmic expression of clock genes. Therefore, we performed macroarray screening of tumour and adjacent tissues in patients undergoing surgery for colorectal carcinoma. We identified 17 miRNAs showing expression that was more than 100 times higher in tumour tissue compared to adjacent tissue. From in silico analysis, miR-34a-5p was selected as showing a computer-predicted interaction with PER2. Real-time PCR revealed a negative correlation between expression of PER2 mRNA and miR-34a in patients with more advanced cancer stage. Expression of miR-34a was up-regulated in cancer tissue compared to adjacent tissue. High miR-34a expression was associated with better survival of patients. miR-34a showed lower expression levels in male patients with lymph node involvement, and a trend towards decreased expression in male patients with distant metastases. Male patients, but not female patients, with high expression of miR-34a and who were free of distant metastases and/or lymph node involvement showed better survival. Therefore, we proposed that expression of miR-34a was regulated in a sex-dependent manner and could be considered a marker of prognosis in earlier cancer stages in male patients.

Sex-dependent correlation between survival and expression of genes related to the circadian oscillator in patients with colorectal cancer.

Recent evidence supports the important role of the circadian system in cancer progression in humans. The aim of the present study is to evaluate clock (cry1, cry2 and per2) and clock-controlled (vascular endothelial growth factor-a, early growth response protein 1 and estrogen receptor ß) gene expression in colorectal cancer and adjacent tissue and identify a possible link between survival of patients and expression of above mentioned genes. The study includes 64 patients of both sexes with previously diagnosed colorectal cancer. RNA was extracted from the tumor tissue and adjacent parts of the resected colon, and real-time PCR was used for detection of clock gene expression. Expression of cry2 and per2 was significantly downregulated in tumor tissue compared to adjacent tissues. After splitting of the cohort according to sex, we detected downregulated levels of cry2 and per2 in male patients, but not in females. Splitting of male and female sub-cohorts according to presence of metastases revealed significant donwregulation of cry2 expression in female patients without distant metastasis. Better survival rate was associated with low expression of cry2 in female patients. Moreover, we observed an increase in cry1 expression in female patients with distant metastases in tumor compared to adjacent tissue. Accordingly, women with high expression of cry1 in tumor tissue displayed worse survival, which was not observed in men. Taken together, expression of clock and clock-controlled genes in tumors of males and females clustered according to presence of distant metastases correlated with survival analysis. Studied clock-controlled genes also showed sex-dependent changes. Low expression of vegf-a in tumor correlated with better survival in men but not in women. High expression of estrogen receptor ß mRNA was related to better survival in women but not in men. Low expression of vegf-a, egr1 and estrogen receptor ß was associated with worse survival in women compared to men. Our data indicate sex-dependent associations between clock and clock-controlled gene expression in cancer tissue and patient's survival prognosis.

Expression of cell cycle regulatory factors hus1, gadd45a, rb1, cdkn2a and mre11a correlates with expression of clock gene per2 in human colorectal carcinoma tissue.

Deregulated expression of clock gene per2 has previously been associated with progression of cancer. The aim of the present study was to identify genes related to per2 expression and involved in cell cycle control. Patients surgically treated for colorectal carcinoma with up-regulated and down-regulated per2 expression in cancer versus adjacent tissue were studied. Total RNA from cancer tissue of these patients was used to specify genes associated with altered per2 expression using the Human Cell Cycle RT(2) profiler PCR array system. We identified seven genes positively correlated (hus1, gadd45α, rb1, cdkn2a, cdk5rp1, mre11a, sumo1) and two genes negatively correlated (cdc20, birc5) with per2 expression. Expression of these seven genes was subsequently measured by real time PCR in all patients of the cohort. Patients were divided into three groups according to TNM classification. We observed an increase in gene expression in cancer tissue compared to adjacent tissue in the first group of patients in all genes measured. Expression of genes positively associated with per2 gene expression was dependent on tumor staging and changes were observed preferentially in cancer tissue. For genes negatively associated with per2 expression we also detected changes in expression dependent on tumor staging. Expression of cdc20 and birc5 was increasing in the proximal tissue and decreasing in the cancer tissue. These results implicate functional involvement of per2 in the process of carcinogenesis via newly uncovered genes. The relevancy of gene expression for determination of diagnosis and prognosis should be considered in relation to tumor staging.

The impact of body position on intra-abdominal pressure measurement: a multicenter analysis.

OBJECTIVE: Elevated intra-abdominal pressure (IAP) is a frequent cause of morbidity and mortality among the critically ill. IAP is most commonly measured using the intravesicular or "bladder" technique. The impact of changes in body position on the accuracy of IAP measurements, such as head of bed elevation to reduce the risk of ventilator-associated pneumonia, remains unclear. DESIGN: Prospective, cohort study. SETTING: Twelve international intensive care units. PATIENTS: One hundred thirty-two critically ill medical and surgical patients at risk for intra-abdominal hypertension and abdominal compartment syndrome. INTERVENTIONS: Triplicate intravesicular pressure measurements were performed at least 4 hours apart with the patient in the supine, 15 degrees , and 30 degrees head of bed elevated positions. The zero reference point was the mid-axillary line at the iliac crest. MEASUREMENTS AND MAIN RESULTS: Mean IAP values at each head of bed position were significantly different (p < 0.0001). The bias between IAPsupine and IAP15 degrees was 1.5 mm Hg (1.3-1.7). The bias between IAPsupine and IAP30 degrees was 3.7 mm Hg (3.4-4.0). CONCLUSIONS: Head of bed elevation results in clinically significant increases in measured IAP. Consistent body positioning from one IAP measurement to the next is necessary to allow consistent trending of IAP for accurate clinical decision making. Studies that involve IAP measurements should describe the patient's body position so that these values may be properly interpreted.

The effect of different reference transducer positions on intra-abdominal pressure measurement: a multicenter analysis.

OBJECTIVE: To investigate the effect of different reference transducer positions on intra-abdominal pressure (IAP) measurement. Three reference levels were studied: the symphysis pubis; the phlebostatic axis; and the midaxillary line at the level of the iliac crest. DESIGN: Prospective cohort study. SETTING: The intensive care units of participating hospitals PATIENTS AND PARTICIPANTS: One hundred thirty-two critically ill patients at risk for intra-abdominal hypertension (IAH). INTERVENTIONS: In each patient, three sets of IAP measurements were obtained in the supine position, using the different reference levels. The IAP measurements obtained at the different reference levels were compared using a paired t-test and Bland-Altman statistics were calculated. MEASUREMENTS AND RESULTS: IAP(phlebostatic) (9.9 +/- 4.67 mmHg) and IAP(pubis) (8.4 +/- 4.60 mmHg) were significantly lower that IAP(midax) (12.2 +/- 4.66 mmHg; p < 0.0001 for both comparisons). The bias between the IAP(midax) and IAP(pubis) was 3.8 mmHg (95% CI 3.5-4.1) and 2.3 mmHg (95% CI 1.9-2.6) between the IAP(midax) and the IAP(phlebostatic). The precision was 3.03 and 3.40, respectively. CONCLUSIONS: In the supine position, IAP(midax) is higher than both IAP(phlebostatic) and IAP(pubis), differences found to be clinically significant; therefore, the symphysis pubis or phlebostatic axis reference lines are not interchangeable with the midaxillary level.

Deregulated expression of the per2 gene in human colorectal carcinoma.

The circadian system is involved in the control of cell proliferation and apoptosis. The aim of this study was to analyze expression of the human per2 gene in patients who underwent surgery for colorectal carcinoma. The study included 25 patients of both genders. Patients were exposed to light from 6:00 a.m. until 9:00 p.m. according to standard hospital practice. Tissue samples were taken from the tumor as well as from the proximal and distal areas of the resected colon at the time of surgery. Surgery was performed during the morning hours. Expression of per2 mRNA was measured by real-time PCR. There was a significant negative correlation between per2 gene expression and tumor staging. Expression of per2 mRNA did not correlate with whether the tumor was localized in the colon or rectum. In comparison with ectomized tissue without malignancy from patients with colorectal carcinoma, our data demonstrate per2 mRNA deregulation in tumor tissue, and suggest a way in which the circadian system can influence tumorigenesis.