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کاربرد نوع شرط:
- جایگاه : پژوهشی
- مجله: Iranian Journal of Biotechnology
- نوع مقاله: Journal Article
- کلمات کلیدی: eCG, Ouchterlony,Polyoclonal Antibody,Solid-Phase Extraction (SPE)
- چکیده:
- چکیده انگلیسی: Background: Equine chorionic gonadotropin (eCG) is commonly used in association with a progestagen treatment to synchronize estrus of goats and ewes during breeding and non-breeding seasons. Classical purification of the eCG from serum includes pH fractionation with metaphosphoric acid two ethanol precipitation steps as well as dialysis followed by fixed-bed chromatography. Objectives: The aim of the current study was to develop an accurate and fast method for production and purification of eCG using polyoclonal antibody assay. Materials and Methods: The blood samples (300 ml) were taken from jugular vein of 17 mares on days 50, 70 and 90 of pregnancy. Plasma of the samples was siphoned and phenol solution has been added to the plasma and stored in the refrigerator until eCG extraction. To prepare polyoclonal antibody against eCG, four male rabbits, about 4-months-old and 2-kg weights, were chosen. A basic immunization was done by injecting 25 IU of eCG to the rabbits. Ouchterlony assay or double immunodiffusion test was used to assess the immunization and the titer of antiserum against eCG. eCG has been purified from the plasma via Solid-Phase Extraction (SPE) method. Results: Results based on agarose-gel double immunodiffusion test showed that rabbits have been completely immunized. SDS-PAGE analysis showed purified eCG is extracted without any significant contamination. Conclusions: The extraction of eCG with polyclonal antibody using SPE method and production of anti- eCG antiserum in rabbits is suitable and may be a cost effective method for large scale production of eCG and anti- eCG antiserum in Iran.
- انتشار مقاله: 09-07-1392
- نویسندگان: Mohammad Ali Sharif,Hamid Kohram,Ahmad Zare Shahneh,Hossein Zolfagharian,Bahman Abedi Kiasari,Mehdi Hedayati
- مشاهده
- جایگاه : پژوهشی
- مجله: International Journal of Advanced Biological and Biomedical Research
- نوع مقاله: Journal Article
- کلمات کلیدی: Apoptosis,Thyroid cancer,Carcinoma,NVP-BEZ235,PD-332991
- چکیده:
- چکیده انگلیسی: Introduction
Thyroid cancer is a malignant disease with poor prognosis. The PI3K/Akt/mTOR and Cyclin-Dependent Kinase 4 (CDK4) pathways are vital regulators of tumor cell proliferation and survival. Therefore the present study was designed to use dual inhibition of such pathways to kill thyroid cancer cells.
Methods and materials
The effects of each inhibitors on human ATC and BCPAP cell lines were evaluated by MTT assay. The suitable concentrations of inhibitors were determined and synergistic effects of such inhibitors were evaluated by bax/bcl-2 mRNA ratio, Caspase-3, and Caspase-9 activity assay as well as Akt, mTOR, and CDK4.
Results:
Our finding showed that both ATC and BCPAP cell proliferation is significantly inhibited by PD-332991(PD) and NVP-BEZ235 (NVP) in a time and concentration-dependent manner (P- انتشار مقاله: 05-04-1398
- نویسندگان: Neda Hamidipour,Mehdi Fazeli,Mehdi Hedayati,Mohsen Dehghani,Razieh Gerami
- مشاهده
- جایگاه : پژوهشی
- مجله: International Journal of Advanced Biological and Biomedical Research
- نوع مقاله: Journal Article
- کلمات کلیدی: Apoptosis,Thyroid cancer,Carcinoma,NVP-BEZ235,PD-332991
- چکیده:
- چکیده انگلیسی: Introduction
Thyroid cancer is a malignant disease with poor prognosis. The PI3K/Akt/mTOR and Cyclin-Dependent Kinase 4 (CDK4) pathways are vital regulators of tumor cell proliferation and survival. Therefore the present study was designed to use dual inhibition of such pathways to kill thyroid cancer cells.
Methods and materials
The effects of each inhibitors on human ATC and BCPAP cell lines were evaluated by MTT assay. The suitable concentrations of inhibitors were determined and synergistic effects of such inhibitors were evaluated by bax/bcl-2 mRNA ratio, Caspase-3, and Caspase-9 activity assay as well as Akt, mTOR, and CDK4.
Results:
Our finding showed that both ATC and BCPAP cell proliferation is significantly inhibited by PD-332991(PD) and NVP-BEZ235 (NVP) in a time and concentration-dependent manner (P- انتشار مقاله: 05-04-1398
- نویسندگان: Neda Hamidipour,Mehdi Fazeli,Mehdi Hedayati,Mohsen Dehghani,Razieh Gerami
- مشاهده
- جایگاه : پژوهشی
- مجله: Asian Pacific Journal of Cancer Prevention
- نوع مقاله: Journal Article
- کلمات کلیدی: Anaplastic thyroid carcinoma,CD133 antigen,Phosphoinositide-3-kinase,Sodium-iodide symporter
- چکیده:
- چکیده انگلیسی:
Background: Thyroidectomy, radioactive iodine therapy, chemotherapy, or their combination are treatments of choice for thyroid cancers. However, cancer stem cells (CSCs) may become resistant to therapy, and mutations in somatic genes affect radioiodine uptake. This study determined the effect of a phosphoinositide-3-kinase (PI3K) inhibitor on anaplastic thyroid CSCs. Materials and Methods: The magnetic-activated cell sorting assay was used for segregating CD133-positive CSCs from three anaplastic thyroid carcinoma (ATC) cell lines (C643, SW1736, and 8305C). After confirming the cells’ purity by flow cytometry, they were treated with 5, 10, 20, or 25 μM LY294002, a PI3K inhibitor, and then evaluated at 24 and 48 h. The sodium-iodide symporter (NIS) mRNA level was determined using the quantitative real-time polymerase chain reaction. NIS protein expression was evaluated using western blotting. Results: The PI3K inhibitor, at different concentrations and times, increased the NIS mRNA level (1.30-6.17-fold, P < 0.0001). If the NIS mRNA level in LY294002-treated CD133-positive CSCs was increased more than 2-fold, the NIS protein content was detectable. Conclusions: CD133-positive CSCs isolated from ATC cell lines expressed NIS mRNA and protein after PI3K inhibition. Our findings suggest that molecularly targeted CSC therapy may improve the treatment efficacy of aggressive cancers like ATC.- انتشار مقاله: 27-03-1396
- نویسندگان: Farzaneh Bozorg-Ghalati,Mehdi Hedayati,Mehdi Dianatpour,Fereidoun Azizi,Nariman Mosaffa,Davood Mehrabani
- مشاهده
- جایگاه : پژوهشی
- مجله: Asian Pacific Journal of Cancer Prevention
- نوع مقاله: Journal Article
- کلمات کلیدی: Medullary thyroid carcinoma,RET proto-oncogene,Calcitonin,microRNA-323,FFPE
- چکیده:
- چکیده انگلیسی:
Background: Medullary thyroid cancer (MTC) is an endocrine tumor featuring parafollicular or C-cell differentiation, with calcitonin as a specific biomarker in MTC diagnosis. Germline mutations in the RET proto-oncogene are considered responsible for its familial occurrence and somatic mutations can cause sporadic lesions. MicroRNAs can act as oncogenes or tumor suppressors by inhibiting the expression of target genes.. The aim of this study was to investigate relationships between plasma levels of calcitonin and miRNA323 expression in MTC patients with or without RET mutation. Methods: In this cross-sectional study, MTC lesions (based on pathological confirmation) were investigated. Genomic DNA was extracted and Exons 10 and 11 of RET were genotyped using PCR-sequencing. Division was into two groups of 43 cases each with or without mutation. Plasma levels of calcitonin were determined in both. Results: miRNA323 was measured using real-time-PCR. After performing normality tests, independent T-tests and Mann Whitney tests were used for the statistical comparison of parametric and nonparametric data, respectively. Plasma levels of calcitonin were significantly higher in MTC cases without a RET mutation compared to those with a mutation. Conclusion: There was no significant difference between the two groups regarding the expression of miRNA323 so that this parameter could not be used as a bio-index germ line mutations in MTCs. However, determination of calcitonin levels in plasma might be helpful in this regard.- انتشار مقاله: 02-02-1396
- نویسندگان: Samira Ehyaei,Mehdi Hedayati,Marjan Zarif Yeganeh,Sara Sheikholeslami,Mahsa Ahadi,Sayed Asadollah Amini
- مشاهده