Induction of Various Pro-inflammatory Mediators and Their Receptors in Human Astrocytoma Cell Line Stimulated by beta-amyloid Protein

BACKGROUND: The induction of production and production inhibition of alpha1-antichymotrypsin (ACT), IL-1 alpha, IL-1 alpha receptor and macrophage inflammatory protein-1 (MIP-1) receptor in A beta1-42 (A beta)-stimulated U373MG cell, the human astrocytoma cell line, have never been reported. METHODS: U373MG cells (1 x 10(6) cells in RPMI-1640 media) were incubated for overnight after administration of a single dose of 20 micro M of A beta or 0.5 ng/ml of TNF alpha or both. Actinomycin D (2.5 micro M) or cycloheximide (2.5 micro M) was also added to the cell suspension. Messenger RNA expression of ACT, IL-1 alpha, IL-1 alpha receptor and MIP-1 receptor was measured by RT-PCR. Western blot was done and nitrocellulose paper was stained with anti-ACT and anti-GFAP antibody. NF kappa B activation after treatment of A beta in U373MG cells was detected by electrophoretic mobility-shift assay. RESULTS: A beta and TNF alpha both increased production of ACT in a dose-dependent manner. TNF alpha enhanced A beta-induced mRNA had increases of ACT, IL-1 alpha, IL-1 alpha receptor and MIP-1 alpha receptor. Activated NF kappa B was demonstrated in the A beta, TNF alpha-stimulated U373MG cells. Actinomycin suppresses mRNA level of ACT and IL-1 alpha receptor but cycloheximide inhibits the expression of ACT, IL-1 alpha and MIP-1 alpha receptor. CONCLUSIONS: TNF alpha increases synthesis of ACT, IL-1 alpha, IL-1 alpha receptor and MIP-1 alpha receptor in A beta-stimulated astrocyte, which, as a result, may contribute to the neuroinflammation of Alzheimer's disease.

Apolipoprotein E Gene and Alpha 1-Antichymotrypsin Gene in Brain of Sporadic Alzheimer's Disease

BACKGROUND: This study aims to detect any causative genetic alterations and to demonstrate any correlations of these genes in the pathogenesis of mostly late-occurring sporadic type of Alzheimer's disease (AD). METHODS: A total of 67 registered cases of autopsy-confirmed brain tissues were analyzed. Included here was sporadic AD (n=41), vascular dementia (n=17), and non-demented physiologically aging control brains (n=9). ApoE genotyping was done with the enzymatic digestion, and allele specific PCR was done to analyze the -491 A/T polymorphism of ApoE. Detection of polymorphism of alpha 2-macroglobulin (A2M) was done with enzymatic digestion and DNA sequencing. RT-PCR products were electrophoresed to detect mRNA expression of alpha 1-antichymotrypsin (ACT). RESULTS: A prevalence rate of ApoE E4 genotype (E3/E4, E4/E4) showed significantly higher in patients with AD than in patients with vascular dementia (43.8% vs. 11.7%, p=0.019). Only 1 out of 4 cases of sporadic AD was associated with the E4/E4 allele. -491A/ T polymorphism of the ApoE promoter was found only in AD (2/41 cases, 4.9%). The incidence of heterozygous allelic polymorphism with 5 bp deletions in exon 18 of A2M-2 was 4.9% (2 out of 41) in AD. Messenger RNA expression of ACT, which is closely associated with the ApoE E4 allele, was increased in AD in comparison with normal control (p=0.0002). CONCLUSIONS: ApoE4 genotype and ACT are closely related to the pathogenesis of late-onset sporadic AD. Neither -491 polymorphism of ApoE promoter nor A2M-2 showed close association with AD in these brain samples.

Relation of alpha-1-antichymotrypsin and p53 Protein Expression to Bony Metastasis in Advanced Prostate Cancer / 대한비뇨기과학회지

PURPOSE: The purpose of this study was to investigate whether the expressions of alpha-1 -antichymotrypsin(ACT) and/or p53 protein in advanced prostate cancer were related with bony metastases. MATERIALS AND METHODS: The immunohistochemical study with ACT and p53 included 7 archival transurethral resection and 14 prostate biopsy specimens from patients with prostate cancer who showed high serum PSA level(>10ng/ml) and periprostatic or lymph node involvement on imaging study. Whole body bone scan was perfomed to detect bony metastatis in all patients. RESULTS: Four out of 5 cases showing strong expression patterns with ACT protein showed abnormal hot uptakes on whole body bone scan. ACT proteins were weakly expressed in seven out of eight cases without bony metastases. p53 protein was expressed in 13 cases, but there was no statistically significant relation between the expression of p53 protein and bony metastasis. Either, there was no significant relation between ACT and p53 protein expression patterns. CONCLUSIONS: Our results suggested that the strong expression of ACT protein combined with high serum PSA level(>10ng/ml) and whole body bone scan could be the useful method for confirming bony metastasis although not adequate for screening test. The expression of p53 protein appears to be associated with progression of prostate cancer, but there was no statistically significant relation with bony metastasis.

Significance of Alpha-1-antichymotrypsin in Prostate Cancer / 대한비뇨기과학회지

Predominant type of serum prostate specific antigen (PSA) is a complexed-form which is bound to alpha-1-antichymotrypsin (ACT). Major fraction of serum ACF is derived from the liver and ACT has recently been demonstrated to be produced by PSA-producing prostatic epithelium as well. However, the feature and significance of prostate-derived ACT remain ill-defined. We herein immunohistochemically studied prostatic tissues in 40 patients with prostate cancer and 20 patients with benign prostatic hyperplasia (BPH) using antibody to ACT, cytokeratin AE3 and chromogranin A. In normal portion of prostate, secretory epithelia of central zone and peripheral zone, and basa1 cells and neuroendocrine cells of peripheral zone showed positive staining for ACT. Benign hyperplastic prostatic gland did not stain for ACT. On the study of prostate cancer tissues, relatively increased staining for ACT were found in solid and infiltrative type of cancer cells and high grade cancer cells. High intensity staining for ACT were observed in normal prostatic tissues adjacent to invasive cancer cells. In conclusions, basal cells as well as secretory epithelia of the normal prostate gland may be the source of serum ACT production by prostate cancer may be closely related with its malignant and invasive potential.

Dedifferentiated Liposarcoma of the Retroperitoneum: A case report

We report a case of dedifferentiated liposarcoma of retroperitoneum as a recurrent form in a 41 year old male. The patient received a extirpation for retroperitoneal mass and diagnosed as myxoid liosarcoma 4 years ago. The patient experienced 3 recurrences over a period of 4 years and diagnosed as myxoid liposarcoma in the second, third recurrence also. Histologically, the mass was composed of several clearly distinct elements : well differentiated liposarcoma, myxoid liposarcoma, myxoid malignant fibrous histiocytoma, poorly differntiated sarcoma, and fibrosarcoma. Immunohistochemically, S-100 protein was expressed in the area of spindle cell sarcoma, well differentiated liposarcoma, and malignant fibrous histiocytoma but alpha-1-antichymotrypsin was only expressed in the area of myxoid malignant fibrous histiocytoma.