Human XIAP(X-linked inhibitor of apoptosis) ELISA Kit

L-asparagine essentially regulates growth and proliferation of cancer cells. L-asparaginase is an anticancer enzyme that deprives the cancer cells of L-asparagine. The purpose of this study was to explore the mechanism of a novel L-asparaginase from Pseudomonas fluorescens on L-asparagine deprivation mediated anti-proliferation, apoptosis in human gastric adenocarcinoma cells and to evaluate inhibition of angiogenesis. We observed that, the presence of extracellular L-asparagine was essential for the growth of AGS cells. L-asparagine deprivation by L-asparaginase induced metabolic stress, cytotoxicity and apoptosis by GO phase cell-cycle arrest, modulated the mitochondrial membrane integrity, accelerated caspase-3 activation and instigated DNA damage. The RT-PCR analysis of pro-apoptosis genes: bak1, bax, bbc3, bik, pmaip1, bnip3l, apaf1, casp3, casp7 and casp9 were significantly higher (P < 0.05), while anti-apoptotic markers xiap, bid, mcl1, and death receptor genes tnf and tradd were significantly down-regulated (P < 0.05). Additionally, higher protein expressions of p53, caspase-3 and TEM analysis showing modulations in mitochondria confirmed intrinsic apoptosis pathway. The enzyme impeded tumor progression through inhibition of cell migration and vascular remodelling of endothelial cells. Our findings suggests that the action of L-asparaginase alters mitochondrial membrane permeability and auxiliary activates intrinsic apoptosis. Therefore, this mechanistic approach might be considered as a targeted enzymotherapy against gastric adenocarcinoma. (C) 2018 Elsevier Inc. All rights reserved.

The accumulation of alpha-syn which induce endoplasmic reticulum stress (ERS) and mediate various signaling pathways involved in DA neuronal degeneration, and the apoptosis of dopamine (DA) neurons are pathological markers of Parkinson's disease (PD). High-temperature requirement protein A2 (HtrA2) is synthesized in the endoplasmic reticulum, and the expression level of HtrA2 can be upregulated by drugs or by unfolded proteins. Ucf-101 is a specific inhibitor of HtrA2, and studies have shown that Ucf-101 reduced apoptosis in PC12 cells. Our study showed that PC12 cells treated with 60 mu M 6-OHDA for 24 h had significantly decreased cell viability compared to that of controls. A low concentration (2.5 mu M) of Ucf-101 decreased the apoptosis rate of the PD cell model, but a high concentration (>= 10 mu M) increased the apoptosis rate, compared to that of controls. 6-OHDA upregulated the expression of HtrA2, alpha-syn, CHOP, Grp78 and active caspase-3 and reduced the levels of TH and XIAP. Ucf-101 reduced the level of ERS and apoptosis both in vivo and in vitro. The ratio of p-GSK3 beta (Tyr216 to Ser9) increased in PD rats. However, Ucf-101 down-regulated the activation of GSK3 beta and activated the Wnt/beta-catenin pathway that was caused by 6-OHDA. Ucf-101 activated the Wnt/beta-catenin pathway and significantly attenuated 6-OHDA-induced neurotoxicity, which was related to the inhibition of ERS and the reduction of the apoptosis rate of PC12 cells and DA neurons in the midbrain of PD rats. Ucf-101 has certain neuroprotective effects. (C) 2019 Elsevier Ltd. All rights reserved.