[Detection of murine tissues that express opticin]

The LRR [leucine rich proteoglycans] is a molecular recognition motif found in proteins with some roles in cell adhesion, signal transduction, DNA repair and RNA processing. Opticin is a member of this family. Takanosu et al [2001] detected messenger RNA expression of mouse opticin in the eye, heart, brain, testis, thyroid and epididymis by dot blot hybridization. In this study, expression levels of mRNA and protein of opticin was investigated by two monoclonal antibodies which were raised against opticin peptides. By this method structure of opticin in human and mouse has been studied. Mouse tissues including, kidney, testis, liver, lung, heart, brain, muscle, spleen and eye were isolated. Opticin expression was identified at mRNA and protein levels by RTPCR and Western blot. PCR analysis revealed that opticin mRNA is expressed in all the tissues studied except for the lung. However, opticin protein was detected in all tissues analyzed. Expression of opticin in the adult murine tissues may suggest functions other than that of putative regulation of vitreous collagen fibrillogenesis for this molecule

[Analysis of gene and protein expression of opticin in human cancer cell lines]

Opticin, as a member of small leucine rich proteoglycans [SLRPs], mainly expressed in human eyes. It is also expressed in human brain, ligament, liver and skin, but at lower levels than the eyes. In the present study, expression of opticin was studied in a variety of human cancer cell lines. In this basic study, human cancer cell lines including, A-172 [brain cancer], T47D [mammary gland carcinoma], Paca-2 [pancreas carcinoma], Ej-138 [bladder carcinoma], Calu6 [lung carcinoma], ACHN [kidney cancer], SKOV3 [ovarian cancer], LS-180 [colon carcinoma], A-375 [skin cancer], PC3 [prostate adenocarcinoma], CLL-CII [chronic lymphocytic leukemia] were cultured under standard conditions. Opticin expression in the cell lines was detected by RT-PCR and Western blot. All cancer cell lines, except A-172, expressed opticin at both mRNA and protein levels. In the current study, human cancer cell lines were shown to express opticin whereas previous studies did not detect opticin in normal tissues. This finding may represent opticin as a novel tumor marker in a wide variety of cancers and future studies can be performed to evaluate function of this protein in carcinogenesis

Modeling breast acini in tissue culture for detection of malignant phenotype reversion to non-malignant phenotype

Evidence is accumulating to support disruption of tissue architecture as a powerful event in tumor formation. For the past four decades, intensive cancer research with the premise of "cancer as a cell based-disease" focused on finding oncogenes or tumor suppressor genes. However, the role of the tissue architecture was neglected. Three dimensional [3D] cell cultures which can recapitulate major aspects of the microenvironment are appropriate models for exploring cancer. For the first time in Iran, we have launched Matrigel based non-malignant, tumorigenic and reverted breast 3D cell cultures. Non-tumorigenic MCF-10A and tumorigenic MCF-7 breast cell lines were cultured on plastic and Matrigel. MCF-7 cell lines were reverted to normal phenotype via AIIB2 and LY 294002 inhibitors against beta1 integrin and class I phosphatidylinositol 3-kinase, respectively. MCF-10A acini were distinguishably different from MCF-7 on Matrigel. MCF-10A formed organized hollow spherical structures which were in stark contrast to the MCF-7 disorganized cluster of cells. Matrigel allowed visual monitoring of MCF-7 cells treated with inhibitors. After treatment of MCF-7 cells, we observed reversion of MCF-7 phenotype toward normal, comparable to MCF-10A acini. The 3D culture provides a microenvironment which allows malignant and non-malignant cells to demonstrate near physiological behavior and this can distinguish nonmalignant from malignant cells. The 3D culture also allows visual monitoring of malignant phenotype reversion to organized spheres

Anti-zona pellucida antibodies in infertile patients in relation to multiple puncture of ovaries and unexplained infertility

Auto antibodies to zona-pellucida [AZA] seem to be important autoantibodies implicated in reproduction, with substantial role in both endocrine and reproductive functions of the human ovary. There are some debates on the relation of AZA with infertility, repeated In Vitro Fertilization [IVF] attempts, and outcome of it. In this study, we assessed the presence of AZA in the follicular fluids [FFs] of women who underwent intra cytoplasmic sperm injection [ICSI], in relation to etiology of infertility and multiple puncture of ovaries. In this prospective study, follicular fluids were evaluated from 96 infertile women, [19-40 years old, 31.5 +/- 5.1], who were c and idates for ICSI based on the etiology of infertility. From these 80 women had explained infertility whereas 16 had unexplained infertility. All FFs were evaluated for presence of AZA by ELISA test. Twenty patients [20.8%] were positive for AZA in follicular fluid. In patients with unexplained infertility, AZA antibody in follicular fluid, was significantly higher than the group with proven etiology of infertility [p=0.001]. In addition, 20.4% of patients who had been punctured previously showed AZA in their FFs which is statistically similar to the patients who were punctured for the first time. The high incidence of AZA in infertile women, especially women with unexplained infertility has to be considered. Relation of the presence AZA and repeated puncture of ovaries is still debatable. Determinations of AZA are highly recommended in evaluation of infertile couples especially in patient with unexplained infertility