Preliminary exploration of the expression of acetylcholinesterase in normal human T lymphocytes and leukemic Jurkat T cells.

The classic enzymatic function of acetylcholinesterase (AChE) is the hydrolysis of acetylcholine (ACh) in the neuronal synapse. However, AChE is also present in nonneuronal cells such as lymphocytes. Various studies have proposed the participation of AChE in the development of cancer. The ACHE gene produces three mRNAs (T, H and R). AChE-T encodes amphiphilic monomers, dimers, tetramers (G1 A, G2 A and G4 A) and hydrophilic tetramers (G4 H). AChE-H encodes amphiphilic monomers and dimers (G1 A and G2 A). AChE-R encodes a hydrophilic monomer (G1 H). The present study considered the differences in the mRNA expression (T, H and R) and protein levels of AChE, as well as the molecular forms of AChE, the glycosylation pattern and the enzymatic activity of AChE present in normal T lymphocytes and leukemic Jurkat E6-1 cells. The results revealed that AChE enzymatic activity was higher in normal T lymphocytes than in Jurkat cells. Normal T cells expressed AChE-H transcripts, whereas Jurkat cells expressed AChE-H and AChE-T. The molecular forms identified in normal T cells were G2 A (5.2 S) and G1 A (3.5 S), whereas those in Jurkat cells were G2 A (5.2 S), G1 A (3.5 S) and G4 H (10.6S). AChE in Jurkat cells showed altered posttranslational maturation since a decrease in the incorporation of galactose and sialic acid into its structure was observed. In conclusion, the content and composition of AChE were altered in Jurkat cells compared with those in normal T lymphocytes. The present study opened new avenues for exploring the development of novel therapeutic strategies against T-cell leukemia and for identifying potential molecular targets for the early detection of this type of cancer.

Changes in the Acetylcholinesterase Enzymatic Activity in Tumor Development and Progression.

Acetylcholinesterase is a well-known protein because of the relevance of its enzymatic activity in the hydrolysis of acetylcholine in nerve transmission. In addition to the catalytic action, it exerts non-catalytic functions; one is associated with apoptosis, in which acetylcholinesterase could significantly impact the survival and aggressiveness observed in cancer. The participation of AChE as part of the apoptosome could explain the role in tumors, since a lower AChE content would increase cell survival due to poor apoptosome assembly. Likewise, the high Ach content caused by the reduction in enzymatic activity could induce cell survival mediated by the overactivation of acetylcholine receptors (AChR) that activate anti-apoptotic pathways. On the other hand, in tumors in which high enzymatic activity has been observed, AChE could be playing a different role in the aggressiveness of cancer; in this review, we propose that AChE could have a pro-inflammatory role, since the high enzyme content would cause a decrease in ACh, which has also been shown to have anti-inflammatory properties, as discussed in this review. In this review, we analyze the changes that the enzyme could display in different tumors and consider the different levels of regulation that the acetylcholinesterase undergoes in the control of epigenetic changes in the mRNA expression and changes in the enzymatic activity and its molecular forms. We focused on explaining the relationship between acetylcholinesterase expression and its activity in the biology of various tumors. We present up-to-date knowledge regarding this fascinating enzyme that is positioned as a remarkable target for cancer treatment.

Assessment of Cytogenetic Damage and Cholinesterases' Activity in Workers Occupationally Exposed to Pesticides in Zamora-Jacona, Michoacan, Mexico.

Pesticides have been considered as potential chemical mutagens; however, little is known about toxic and genotoxic effects during pesticide application in Zamora-Jacona, Michoacan State in Mexico. This study sought to determine DNA damage and cholinesterase activities inhibitions in 54 agricultural workers exposed to complex mixtures of pesticides vs. control group (26 individuals) using Comet assay in peripheral whole blood, micronucleus (MN) test in oral mucosa cells, Cytokinesis-blocked MN assay in lymphocytes (L-CBMNcyt) and measuring AChE and BChE activities in whole blood and plasma samples, respectively. Exposed subjects demonstrated significantly elevated levels of primary (Comet assay: tail intensity, tail length, tail moment, Olive tail moment) and permanent DNA damage (MN assay: in blood/buccal cells; frequencies of nuclear buds, binucleated cells, cells with condensed chromatin, karyorrhexis, pyknosis, and karyolysis). However, inhibition of cholinesterase activities (AChE and BChE) was not observed in the workers. Confounding factors including sex, age, BMI, working exposure period, protection level, smoking habit (cigarettes per day units), alcohol consumption (weekly), medication, were considered in the analysis. These combined techniques demonstrated usefulness in the health hazards risks pesticide exposure assessment and suggested the need for periodic monitoring together with the education and the training of occupational workers for the safe application of potentially harmful pesticides.

Over-production of IFN-gamma and IL-12 in AhR-null mice.

The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that mediates toxicity of environmental pollutants such as 2,3,7,8-tetrachlorodibenzo-p-dioxin. The exposure to AhR agonists results in profound suppression of cellular and humoral immune responses and compromises host to infectious disease. Therefore, to define the role of AhR in the immune response, spleen cells from ovalbumin (OVA)-immunized and naïve mice were removed and stimulated in vitro with either OVA or mitogen concanavalin-A (Con A), respectively. Proliferation, CD19+, F4/80+, CD4+ and CD8+ T cells expansion and cytokines production were measured in C57BL/6-AhR-/- mice (AhR-/-) and compared with immune response in similarly immunized age-matched wild type (AhR+/+) mice. In response to OVA immunization, AhR-/- mice had similar levels of serum OVA-specific IgG2a, IgG1, and IgG2b compared with AhR+/+ animals. However, AhR-/- mice showed splenomegalia and an increase in B cells. No changes were observed on proliferation and IL-4 secretion, although AhR-/- cells produced more IFN-gamma and IL-12 than AhR+/+ cells. Similar results were observed with Con A stimulation, a decrease on IL-5 and no change on IL-2 secretion were observed on AhR-/- cells compared with AhR+/+ cells in response to Con A stimulation. High levels of IFN-gamma mRNA were detected in AhR-/- lymphocytes, but IL-4 mRNA levels in AhR-/- cells were similar to those in AhR+/+ mice. These data suggest that AhR may play an important role in the normal development and function of immune system by down-regulating IFN-gamma and IL-12 expression.