MeCP2 is an abundant mammalian protein that binds to methylated CpG. We have found that native and recombinant MeCP2 repress transcription in vitro from methylated promoters but do not repress nonmethylated promoters. Repression is nonlinearly dependent on the local density of methylation, becoming significant at the density found in bulk vertebrate genomic DNA. Transient transfection using fusions with the GAL4 DNA binding domain identified a region of MeCP2 that is capable of long-range repression in vivo. Moreover, MeCP2 is able to displace histone H1 from preassembled chromatin that contains methyl-CpG. These properties, together with the abundance of MeCP2 and the high frequency of its 2 bp binding site, suggest a role as a global transcriptional repressor in vertebrate genomes.

Increasing evidence supports the involvement of cholinesterases in tumorigenesis. Several tumour cells show ChE activity, while the acetyl- (AChE) and butyrylcholinesterase (BuChE) genes are amplified in leukemias, ovarian carcinoma and other cancers. ChE activity was measured in 31 samples of tumoral breast (TB) and 20 of normal breast (NB). Despite the wide variations observed, BuChE predominated over AChE both in TB and NB. The mean AChE activity in NB was 1.61 nmol of the substrate hydrolysed per minute and per miligram protein (mU/mg), which rose to 3.09 mU/mg in TB (p = 0.041). The BuChE activity dropped from 5.24 mU/mg in NB to 3.39 mU/mg in TB (p = 0.002). Glycolipid-linked AChE dimers and monomers and hydrophilic BuChE tetramers and monomers were identified in NB and TB, and their proportions were unmodified by the neoplasia. The amount of AChE forms reacting with wheat germ agglutinin (WGA) decreased in TB while that of BuChE species was unaffected, demonstrating that the glycosylation of AChE was altered in TB. The binding of AChE and BuChE with antibodies was unaffected by the neoplasia. The difference in lectin reactivity between erythrocyte and breast AChE, the lack of AChE in blood plasma, and the finding of monomeric BuChE in breast but not in plasma suggest that breast epithelial cells produce AChE for membrane attachment and hydrophilic BuChE for secretion. Several reasons are provided to explain the altered expression of ChEs in breast cancer.

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The aberrations of cholinesterase (ChE) genes and the variation of ChE activity in cancerous tissues prompted us to investigate the expression of ChEs in colorectal carcinoma. The study of 55 paired specimens of healthy (HG) and cancerous gut (CG) showed that acetylcholinesterase (AChE) activity fell by 32% and butyrylcholinesterase (BuChE) activity by 58% in CG. Abundant AChE-H, fewer AChE-T, and even fewer AChE-R and BuChE mRNAs were observed in HG, and their content was greatly diminished in CG. The high level of the AChE-H mRNA explains the abundance of AChE-H subunits in HG, which as glycosylphosphatidylinositol (GPI)-anchored amphiphilic AChE dimers (G2(A)) and monomers (G1(A)) account for 69% of AChE activity. The identification of AChE-T and BuChE mRNAs justifies the occurrence in gut of A12, G4(H) and PRiMA-containing G4(A) AChE forms, besides G4(H), G4(A) and G1(H) BuChE. The down-regulation of ChEs might contribute to gut carcinogenesis by increasing acetylcholine availability and over-stimulating muscarinic receptors.

Because of the probable involvement of cholinesterases (ChEs) in tumorigenesis, this research was addressed to ascertaining whether breast cancer metastasis alters the content of acetylcholinesterase (AChE) and/or butyrylcholinesterase (BuChE) in axillary lymph nodes (LN). ChE activity was assayed in nine normal (NLN) and seven metastasis-bearing nodes (MLN) from women. AChE and BuChE forms were characterised by sedimentation analyses, hydrophobic chromatography and western blotting. The origin of ChEs in LN was studied by lectin interaction. AChE activity dropped from 21.6 mU/mg (nmol of the substrate hydrolysed per minute and per milligram protein) in NLN to 3.8 mU/mg in MLN (p < 0.001), while BuChE activity (3.6 mU/mg) was little affected. NLN contained globular amphiphilic AChE dimers (G2A, 35%), monomers (G1A, 30%), hydrophilic tetramers (G4H, 8%), and asymmetric species (A4, 23%, and A8, 4%); MLN displayed only G2A (65%) and G1A (35%) AChE forms. NLN and MLN contained G4H (79%), G4A (7%), and G1H (14%) BuChE components. Neither the binding of ChE forms with lectins and antibodies nor the subunit size were altered by metastasis. The higher level of AChE in NLN than in brain and the specific pattern of AChE forms in NLN support its role in immunity. The different profile of AChE forms in NLN and MLN may be useful for diagnosis.