miR-141-3p commonly regulates human UGT1A isoforms via different mechanisms.

UDP-Glucuronosyltransferase (UGT) 1A enzymes catalyze the glucuronidation of various compounds. Since no correlation was observed between the protein and mRNA expression of some UGT1A isoforms in the human liver, the involvement of post-transcriptional regulation was hypothesized. We examined whether microRNAs (miRNAs) regulate human UGT1A, focusing on the predicted miR-141-3p. A luciferase assay revealed that a miRNA recognition element for miR-141-3p in the 3'-untranslated region of UGT1A mRNA was functional. Overexpression of miR-141-3p in HEK293 cells stably expressing UGT1A1, UGT1A3, UGT1A4, UGT1A6, UGT1A7, or UGT1A9 significantly decreased the protein expression of all UGT1As. The mRNA levels of the UGT1As, except for UGT1A9, were also decreased by miR-141-3p. This indicated that the regulation mechanisms by miR-141-3p were different between UGT1A9 and the other UGT1A isoforms. Overexpression of miR-141-3p in HuH-7 or Caco-2 cells significantly decreased 4-methylumbelliferone O-glucuronosyltransferase activities, suggesting that miR-141-3p down-regulates endogenous UGT1As. No correlation was observed between miR-141-3p and the UGT1A mRNA levels in a panel of human liver samples, suggesting that regulation by other factors could hide the contribution of miR-141-3p to the regulation of UGT1A constitutive expression in the human liver. In conclusion, this study revealed that the miR-141-3p is an additional modulator of human UGT1A expression.

Serum microRNA profiles in patients with chronic hepatitis B, chronic hepatitis C, primary biliary cirrhosis, autoimmune hepatitis, nonalcoholic steatohepatitis, or drug-induced liver injury.

PURPOSE: Some blood biomarkers or histological examination by liver biopsy are used for the diagnosis of liver diseases in clinics. However, conventional blood biomarkers show poor specificity and sensitivity, and liver biopsy is highly invasiveness. Therefore, to overcome such disadvantages, specific/sensitive and noninvasive options are desirable. In recent years, circulating microRNAs (miRNAs) have been acknowledged for their potential as disease markers. Actually, several miRNAs have been reported to be biomarker candidates of liver diseases. However, these earlier studies were performed for one disease. Therefore, the specificity as biomarkers was not guaranteed, because they didn't study for the other types of liver injury. In this study, we examined if circulating miRNA could distinguish different types of liver diseases. METHODS: Serum miRNA profiles in 28 patients with chronic hepatitis B, chronic hepatitis C, primary biliary cirrhosis, autoimmune hepatitis, nonalcoholic steatohepatitis or drug-induced liver injury as well as 4 control subjects were determined by TaqMan MicroRNA Array analysis. Principal component analysis (PCA) of selected miRNAs was performed. RESULTS: We identified 37 miRNAs whose levels were significantly different between any of the groups. Although individual miRNAs could not distinguish different types of liver diseases, probably because of similar liver pathology, their profiling by PCA could classify different liver disease groups. CONCLUSIONS: The profiling of the selected miRNAs can be useful to distinguish different types of liver diseases.

Changes in the expression of miRNAs at the pericentral and periportal regions of the rat liver in response to hepatocellular injury: comparison with the changes in the expression of plasma miRNAs.

MicroRNAs (miRNAs) in body fluids have received attention as potential biomarkers of organ damage because miRNAs that are highly or specifically expressed in a given organ are likely released into body fluids as a result of damage to that organ. We previously determined that the plasma miRNA profile in rats was dramatically changed due to acetaminophen (APAP)-induced pericentral necrosis and methapyrilene (MP)-induced periportal necrosis in the liver. The purpose of this study was to examine whether the expression of hepatic miRNAs is differentially modulated at different zones due to injury and to examine the relationship of the hepatic miRNA profile with the changes in the plasma miRNA expression profile. Through the laser microdissection of the periportal and periportal regions of the liver and TaqMan microRNA Array analysis, we found that 49 miRNAs are differentially expressed between the pericentral and periportal regions of control rats. In both APAP- and MP-treated rats, the miRNAs that presented decreased expression dominated in both the injured and non-injured areas compared with the miRNAs that exhibited increased expression. The changes in miRNA expression in each region of the liver were compared with those observed in the plasma. Of the 301 plasma miRNAs with expression that was changed as a result of APAP administration, only 21% were changed in the injured area of the liver. Of the 263 plasma miRNAs with expression that was changed due to MP administration, only 24% were changed in the injured area of the liver. Thus, the miRNA expression profiles in the plasma do not merely reflect the release of miRNAs from the damaged cells in the liver. This report provides the first demonstration of zonal miRNA expression in the liver and of the relationship of the miRNA expression profile in a tissue with the plasma miRNA profile.

Integrated analysis of rifampicin-induced microRNA and gene expression changes in human hepatocytes.

MicroRNAs (miRNAs) post-transcriptionally regulate mRNA expression, controlling global cell function. Altered expression or function of miRNAs causes various diseases. Chemically induced changes in miRNA expression in human tissues are not fully understood. We investigated the changes in miRNA expression by rifampicin, which modulates the expression of various genes related to drug metabolism and pharmacokinetics, in human hepatocytes, and evaluated the relationship with the gene expression changes. We found that 23 miRNAs were increased (>2-fold) and 17 miRNAs were decreased (<0.5-fold) among 150 detected miRNAs, whereas 60 genes were increased and 105 genes were decreased among 22,673 detected genes upon treatment with 10 µM rifampicin. Changes in 17 intragenic miRNAs out of 40 altered miRNAs did not occur in parallel with alterations in their host genes. We searched for the target mRNAs of the miRNAs altered by rifampicin and found that the changes in expression of 16 mRNA/miRNA pairs were inversely associated. Thus, some mRNA expression altered by rifampicin may result from miRNA regulation. In conclusion, we found that rifampicin altered miRNA expression in human hepatocytes. We obtained new insight on the mechanism of the miRNA expression changes and the complicated relationship with gene transcripts.

Cigarette smoking substantially alters plasma microRNA profiles in healthy subjects.

Circulating microRNAs (miRNAs) are receiving attention as potential biomarkers of various diseases, including cancers, chronic obstructive pulmonary disease, and cardiovascular disease. However, it is unknown whether the levels of circulating miRNAs in a healthy subject might vary with external factors in daily life. In this study, we investigated whether cigarette smoking, a habit that has spread throughout the world and is a risk factor for various diseases, affects plasma miRNA profiles. We determined the profiles of 11 smokers and 7 non-smokers by TaqMan MicroRNA array analysis. A larger number of miRNAs were detected in smokers than in non-smokers, and the plasma levels of two-thirds of the detected miRNAs (43 miRNAs) were significantly higher in smokers than in non-smokers. A principal component analysis of the plasma miRNA profiles clearly separated smokers and non-smokers. Twenty-four of the miRNAs were previously reported to be potential biomarkers of disease, suggesting the possibility that smoking status might interfere with the diagnosis of disease. Interestingly, we found that quitting smoking altered the plasma miRNA profiles to resemble those of non-smokers. These results suggested that the differences in the plasma miRNA profiles between smokers and non-smokers could be attributed to cigarette smoking. In addition, we found that an acute exposure of ex-smokers to cigarette smoke (smoking one cigarette) did not cause a dramatic change in the plasma miRNA profile. In conclusion, we found that repeated cigarette smoking substantially alters the plasma miRNA profile, interfering with the diagnosis of disease or signaling potential smoking-related diseases.