SPHK1 (sphingosine kinase 1) induces epithelial-mesenchymal transition by promoting the autophagy-linked lysosomal degradation of CDH1/E-cadherin in hepatoma cells

SPHK1 (sphingosine kinase 1), a regulator of sphingolipid metabolites, plays a causal role in the development of hepatocellular carcinoma (HCC) through augmenting HCC invasion and metastasis. However, the mechanism by which SPHK1 signaling promotes invasion and metastasis in HCC remains to be clarified. Here, we reported that SPHK1 induced the epithelial-mesenchymal transition (EMT) by accelerating CDH1/E-cadherin lysosomal degradation and facilitating the invasion and metastasis of HepG2 cells. Initially, we found that SPHK1 promoted cell migration and invasion and induced the EMT process through decreasing the expression of CDH1, which is an epithelial marker. Furthermore, SPHK1 accelerated the lysosomal degradation of CDH1 to induce EMT, which depended on TRAF2 (TNF receptor associated factor 2)-mediated macroautophagy/autophagy activation. In addition, the inhibition of autophagy recovered CDH1 expression and reduced cell migration and invasion through delaying the degradation of CDH1 in SPHK1-overexpressing cells. Moreover, the overexpression of SPHK1 produced intracellular sphingosine-1-phosphate (S1P). In response to S1P stimulation, TRAF2 bound to BECN1/Beclin 1 and catalyzed the lysine 63-linked ubiquitination of BECN1 for triggering autophagy. The deletion of the RING domain of TRAF2 inhibited autophagy and the interaction of BECN1 and TRAF2. Our findings define a novel mechanism responsible for the regulation of the EMT via SPHK1-TRAF2-BECN1-CDH1 signal cascades in HCC cells. Our work indicates that the blockage of SPHK1 activity to attenuate autophagy may be a promising strategy for the prevention and treatment of HCC.     

Alpha/beta‐hydrolases: A unique structural motif coordinates catalytic acid residue in 40 protein fold families

The alpha/beta‐hydrolases are a family of acid‐base‐nucleophile catalytic triad enzymes with a common fold, but using a wide variety of substrates, having different pH optima, catalyzing unique catalytic reactions and often showing improved chemical and thermo stability. The ABH enzymes are prime targets for protein engineering. Here, we have classified active sites from 51 representative members of 40 structural ABH fold families into eight distinct conserved geometries. We demonstrate the occurrence of a common structural motif, the catalytic acid zone, at the catalytic triad acid turn. We show that binding of an external ligand does not change the structure of the catalytic acid zone and both the ligand‐free and ligand‐bound forms of the protein belong to the same catalytic acid zone subgroup. We also show that the catalytic acid zone coordinates the position of the catalytic histidine loop directly above its plane, and consequently, fixes the catalytic histidine in a proper position near the catalytic acid. Finally, we demonstrate that the catalytic acid zone plays a key role in multi‐subunit complex formation in ABH enzymes, and is involved in interactions with other proteins. As a result, we speculate that each of the catalytic triad residues has its own supporting structural scaffold, similar to the catalytic acid zone, described above, which together form the extended catalytic triad motif. Each scaffold coordinates the function of its respective catalytic residue, and can even compensate for the loss of protein function, if the catalytic amino acid is mutated.     

Synthesis,structure–activity relationship and molecular docking of 3-oxoaurones and 3-thioaurones as acetylcholinesterase and butyrylcholinesterase inhibitors

The present study describes efficient and facile syntheses of varyingly substituted 3-thioaurones from the corresponding 3-oxoaurones using Lawesson’s reagent and phosphorous pentasulfide. In comparison, the latter methodology was proved more convenient, giving higher yields and required short and simple methodology. The structures of synthetic compounds were unambiguously elucidated by IR, MS and NMR spectroscopy. All synthetic compounds were screened for their inhibitory potential against in vitro acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) enzymes. Molecular docking studies were also performed in order to examine their binding interactions with AChE and BChE human proteins. Both studies revealed that some of these compounds were found to be good inhibitors against AChE and BChE.     

Leaf phenology mediates provenance differences in herbivore populations on valley oaks in a common garden

1. Plants from different populations often display a variation in herbivore resistance. However, it is rarely understood what plant traits mediate such differences. 2. It was tested how leaf phenology affects herbivore populations in a 15‐year‐old common garden of valley oaks (Quercus lobata Née) with different populations and maternal parents from throughout the Q. lobata range. 3. The abundance of leaf miners (Stigmella sp. Shrank) and leaf phenology of oaks in the common garden was measured. 4. Leaf miner abundance varied among provenance locations (population), but not among maternal parents within populations. Leaf phenology varied by provenance location and maternal parent, and trees that leafed out earlier accrued higher leaf‐miner abundance. Path analysis indicated that leaf phenology was the likely driver of provenance and parental differences in resistance to leaf miners. 5. Understanding population differences is particularly important when considering transport of genotypes for ornamental or restoration purposes. The present study suggests that similarity in leaf phenology may be one factor that could be used to find genotypes with a similar herbivore resistance to local genotypes.     

Alterations in the hepatic enzymes following experimental lead poisoning

An investigation on the influence of lead toxicity on some of the hepatic enzymes was studied in rats both after a shorter interval of 15 d and after longer intervals of 60 and 90 d. Three different doses of lead as 5, 10, and 50 mg/kg body wt were administered orally on every alternate day. Whereas significant inhibition of succinic dehydrogenase was seen following lead poisoning, the activity acid and alkaline phosphatase increased with lead intoxication. The histoarchitecture of the liver was grossly intact. Liver accumulated less lead compared to kidney at 60 and 90 d.     

Phytoplankton composition and spatial distribution of copper and zinc in the Fal Estuary (Cornwall,UK)

In the estuaries near Falmouth (Cornwall, UK) levels of dissolved copper and zinc are high, due to drainage of copper and tin mines. Phytoplankton species composition in the autumn of 1989 deviated in the metal-contaminated Restronguet Creek from that in other estuarine branches,viz. Fal, Tresillian and Percuil. In the riverine part of Restronguet Creek (Carnon River)Euglena mutabilis, known as an acidophilic (pH 3) metal-resistant flagellate, occurred at micromolar Cu and Zn, whereas in the clean riversChlamydomonas sp. andOocystis sp. occurred at nanomolar Cu and Zn. An ordination analysis revealed the following patterns in Cu, Zn and phytoplankton species composition in the poly- and euhaline waters. Seston-bound Zn and dissolved Zn were in equilibrium,Katodinium rotundatum is Zn-tolerant, andSkeletonema costatum occurred in water with high contents of Cu in seston. However, none of the variables in these patterns correlated at a significant level (p>0.05). The results show that algae-metal interactions are complicated, and that statistical correlations foundin situ need experimental verification.Communication no. 542 of the Delta Institute for Hydrobiological Research.