Rat Pfkl (6-phosphofructokinase, liver type) ELISA Kit

MicroRNAs (miRNAs) affect cancer cell glucose metabolism by targeting mRNAs of diverse enzymes that have been implicated in oxidative phosphorylation (OXPHOS) and glycolytic pathways. However, the mechanisms that underlie miRNA-mediated regulation of phosphofructokinase (PFK), a key rate-limiting enzyme in glycolysis, remain largely unknown. Here, we show that miR-128 directly targets PFK liver type (PFKL) in lung cancer cells and regulates endogenous expression of PFKL at both the mRNA and protein levels. In line with this, overexpression of miR-128 decreased glucose uptake and lactate production, as well as increased cellular ATP content. Interestingly, knockdown of miR-128 was shown to promote lung cancer cell growth and colony formation. Moreover, we observed that miR-128 expression inversely correlated with PFKL mRNA levels in clinic lung cancer samples and that increased PFKL expression predicted poor overall survival in lung cancer patients. Mechanistically, we showed that miR-128 regulates PFKL via a feedback loop that involves inhibition of the AKT signaling pathway. Together, our results suggest that miR-128 acts as a metabolic regulator in lung cancer cells that may be therapeutically exploited.

To better understand the regulatory mechanism of glucose metabolism, sub-adult Genetically Improved Farmed Tilapia (GIFT) were sampled at 0, 1, 3, 8 and 24 h after feeding (HAF) after 36 h of food deprivation, and the time course of changes in plasma nutrients and the transcription of a series of key genes involved with hepatic glucose transport and utilization were analyzed. Compared with the fasting state (0 HAF), plasma glucose increased at 1 HAF, peaked at 3 HAF, decreased at 8 HAF and returned to basal level at 24 HAF. Genes such as glucokinase (gck) and liver type of phosphofructokinase (pfkl) were sensitive indicators for hepatic glycolysis, and their transcription was active at 1-8 HAF, especially at 3 HAF. Genes such as glucose-6-phosphatase catalytic subunit a2(g6pca2) and phosphoenolpyruvate carboxykinase 2 (pck2) were useful markers for hepatic gluconeogenesis, and their transcription was inhibited at 1-3 HAF, especially at 3 HAF. Hepatic gluconeogenesis was up-regulated with feeding time from 3 to 8 h, suggesting a nutritional status transition from anabolism to catabolism. The mRNA level of glycogen synthase 1 (gys1) increased at 3-8 HAF and liver glycogen accumulated at 8 HAF, but the principal expressed gys2 was unexpectedly down-regulated after feeding, suggesting that glycogen storage might be controlled by Gys1 rather than Gys2 in the liver of tilapia. In line with the increased plasma triglyceride and cholesterol concentrations, the transcription of hepatic lipogenic genes including acetyl-CoA carboxylase a (acca), ATP citrate lyase a (aclya) and fatty acid synthase (fas) were all up-regulated during 1-8 HAF (especially 3-8 HAF), suggesting that hepatic lipogenesis was an efficient way to dispose excess glucose in tilapia. Hepatic glucose transport was induced since the feeding time was over 8 h, indicating that hepatic glucose production was already transported to extra-hepatic tissues at 8 HAF, which further confirmed the nutritional status transition at this time. Taking the results together, it was suggested that the physiologically specific time point of postprandial nutritional status transition from anabolism to catabolism occurred at about or a little earlier than 8 HAF in tilapia. (C) 2017 Elsevier B.V. All rights reserved.