lactoferrin and Prostatic-Hyperplasia

The mass spectrometry technique of multiple reaction monitoring (MRM) was used to quantify and compare the expression level of lactoferrin in tear films among control, prostate cancer (CaP), and benign prostate hyperplasia (BPH) groups. Tear samples from 14 men with CaP, 15 men with BPH, and 14 controls were analyzed in the study. Collected tears (2 μl) of each sample were digested with trypsin overnight at 37 °C without any pretreatment, and tear lactoferrin was quantified using a lactoferrin-specific peptide, VPSHAVVAR, both using natural/light and isotopic-labeled/heavy peptides with MRM. The average tear lactoferrin concentration was 1.01 ± 0.07 μg/μl in control samples, 0.96 ± 0.07 μg/μl in the BPH group, and 0.98 ± 0.07 μg/μl in the CaP group. Our study is the first to quantify tear proteins using a total of 43 individual (non-pooled) tear samples and showed that direct digestion of tear samples is suitable for MRM studies. The calculated average lactoferrin concentration in the control group matched that in the published range of human tear lactoferrin concentration measured by enzyme-linked immunosorbent assay (ELISA). Moreover, the lactoferrin was stably expressed across all of the samples, with no significant differences being observed among the control, BPH, and CaP groups.

Topics: Aged; Aged, 80 and over; Amino Acid Sequence; Calibration; Case-Control Studies; Humans; Isotope Labeling; Lactoferrin; Limit of Detection; Male; Middle Aged; Prostatic Hyperplasia; Reproducibility of Results; Tears

To investigate the expression of low-molecular-weight PSA(lw-PSA) and lactoferrin in the expressed prostatic secretion (EPS) from both benign prostatic hyperplasia (BPH) and normal prostate.. Forty human EPS samples obtained from 20 BPH patients and 20 normal males were subjected to two-dimensional gel electrophoresis (2-DE). Mass spectrometry was performed to confirm the nature of the secreted proteins in EPS.. One uniquely expressed protein in BPH was detected and mass spectrometry determined its nature as lw-PSA (molecular weight 10x10(3), pI 8.5-9.3). More importantly, Western blotting analysis also revealed that lw-PSA detected in BPH-EPS, but was undetectable in BPH-free EPS. In addition, up-regulation of Lactoferrin (molecular weight 35x10(3), pI 7-7.5) in BPH-EPS, as compared with BPH-free EPS, was also observed. More interestingly, lactoferrin was absent in prostate cancer tissues.. Our results indicate lw-PSA may be produced specifically by BPH epithelium and it has a potential to be used as a specific biological marker for the diagnosis of BPH. In addition, benign prostatic epithelium can produce more lactoferrin while prostate cancer tissues go without its lactoferrin secretion.

Topics: Aged; Biomarkers; Blotting, Western; Electrophoresis, Gel, Two-Dimensional; Humans; Lactoferrin; Male; Middle Aged; Molecular Weight; Prostate; Prostate-Specific Antigen; Prostatic Hyperplasia; Spectrometry, Mass, Electrospray Ionization

Differences between benign prostatic hyperplasia (BPH) and normal prostate tissue at the level of mRNA expression provide an opportunity to identify candidate genes for this disease. A cDNA subtraction procedure was used to isolate differentially expressed genes in BPH. The subtraction was done by solution hybridization of BPH cDNA against excess normal prostate cDNA. We identified known, EST, and novel genes by sequence and database analysis of the subtracted cDNAs. Several of these cDNAs were used as probes in Northern blotting analysis to confirm over-expression of their corresponding mRNAs in BPH tissues. One highly upregulated sequence of interest shared identity with a known mRNA encoding human NELL2, a protein containing epidermal growth factor-like domains. NELL2 was not previously reported to be expressed in prostate and may code for a novel prostatic growth factor. In situ hybridization analysis of hyperplastic prostate specimens demonstrated that NELL2 mRNA expression is predominantly localized in basal cells of the epithelium. Disease-related changes in the levels of NELL2 may contribute to alterations in epithelial-stromal homeostasis in BPH. (J Histochem Cytochem 49:669-670, 2001)

Topics: Blotting, Northern; DNA, Complementary; Gene Expression Regulation; Humans; In Situ Hybridization; Lactoferrin; Male; Nerve Tissue Proteins; Prostatic Hyperplasia; RNA, Messenger

Using immunoperoxidase procedures, the presence of lactoferrin was investigated in benign hypertrophy (40 cases), differentiated adenocarcinomas (20 cases) and undifferentiated carcinomas (10 cases) of the prostate. In benign hypertrophy, the glandular epithelium and the secretory product were consistently negative. Strong staining for lactoferrin was always observed in neoplastic cells of differentiated adenocarcinomas and in their endoluminal material; in contrast, a very slight positivity was noted in undifferentiated carcinomas. These findings are discussed in relation to the different degree of neoplastic glandular differentiation and functional activity of prostatic carcinomas.

Topics: Adenocarcinoma; Carcinoma; Histocytochemistry; Humans; Immunoenzyme Techniques; Lactoferrin; Lactoglobulins; Male; Prostatic Hyperplasia; Prostatic Neoplasms

Lactoferrin levels have been determined by radialimmunodiffusion in homogenates of both human benign prostatic hypertrophy obtained at open surgery from patients, some of whom had been treated with oestrogens or antiandrogens, and also in prostatic adenocarcinoma tissue removed by transurethral resection. Results show that in untreated benign prostatic hyperplasia there is a statistically lower lactoferrin level in the median compared with the lateral lobes. In patients with benign prostatic hypertrophy treated before prostatectomy with oestrogens or antiandrogens the lactoferrin concentration is decreased. In neoplastic tissue removed by open surgery, the lactoferrin level is very low. Homogenates of tissue resected from prostatic cancer patients show similarly low levels. The concentration of lactoferrin in human prostate is hormone dependent. The role of the protein is considered to be bacteriostatic.

Topics: Adenocarcinoma; Androgen Antagonists; Estrogens; Humans; Hyperplasia; Lactoferrin; Lactoglobulins; Male; Prostate; Prostatic Hyperplasia; Prostatic Neoplasms