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Showing 3 results for Gene Expression
Isolation of IFN- gamma gene from olive baboon and designed semi-quantitative competitive PCR method to quantify gene expression
Dr. K. Azadmanesh, Dr. F. Roohvand, Dr. S. Amini, S. Andalibi Mahmoodabadi, Dr. M. Kazanji,
Volume 10, Issue 3 (10-2004)
Abstract
Background and Aim: Primates have long been used as the most resembling animal model to human. Primates are natural hosts of HTLV/STLV retroviruses. From this family, HTLV-I is a health problem in Khorasan. Recently a new HTLV-I resembling virus has been detected in olive baboon, which suggests this animal as an ideal animal model for therapeutic studies of this disease, for such studies inventing methods and tools to quantify cytokines of this animal, especially IFN-, has high priority. Materials and Methods: Here in, we report methods for isolating cDNA of IFN- from peripheral blood, comparison of its sequence with other primates, construction of competitive plasmids and primers for quantifying this cytokine. In order to standardize the results of quantification, we needed to measure the expression level of a house keeping gene too, so we designed a competitor plasmid and set of primers to quantify the expression of Glycer Aldehyde 3 Phosphate DeHydrogenase (G3PDH) too. Results: Two new cDNA sequences of baboon IFN- and G3PDH were registered in NCBI GENBANK. The competitive PCRs of a sample of this animal measured 7 pg and 0.455 pgof IFN- and G3PDH transcripts respectively. Conclusion: Phylogenetic analysis of IFN- well classified this baboon among old world monkeys, while the structure of this cytokine seems to be same as human. According to the methodology presented here, it is possible to use the ratio of IFN-/G3PDH expression as a tool to compare levels of IFN- production in different samples.
8 Weeks of Resistance Training Effect on Myostatin Gene Expression of Myocardium in Healthy Male Wistar Rats
Amir Rashidlamir, Mohammad Reza Basami, Seyyed Reza Attarzadeh Hosseini, Keyvan Hejazi, Seyyed Mohamad Motevalli Anberani,
Volume 22, Issue 2 (4-2016)
Abstract

Aims: Based on the clinical studies, there are higher myostatin gene expression and serum level in cardiovascular patients than the healthy persons. In addition, based on some studies, regular resistance exercises is in line with a reduction in the myostatin gene expression. The aim of this study was to investigate the effects of 8-week resistance exercises on the myostatin gene expressions of myocardium in the healthy male Wistar rats. 

Materials & Methods: In the experimental study, 13 male adult Wistar rats, aged 15 weeks, were divided into experimental (n=7) and control (n=6) groups. In experimental group, 8-week exercises were done as climbing up a 26-step 1-meter ladder, inclined 85°, with a sinker. (There were 3 sessions a week exercises; each session consisted of 3 sets of 5 repetitions.) 24 hours after the last exercise session and after 12-hour fasting, the rats being dissected, muscle tissue samples were prepared to assess mRNA levels. Data was analyzed in SPSS 16 software using independent T test.

Findings: After 8-week resistance exercises, there was no significant difference in the myostatin mRNA expression level in the myocardium between experimental (1.56±0.44) and control (1.94±0.55) groups (p=0.197).

Conclusion: 8-week resistance exercises leads to no significant change in the myostatin gene expression of myocardium in the healthy Wistar male rats.


Investigation of the Effect of Lactobacillus Brevis Bacteria on the Expression of Rel A, IKB, and Casp3 Genes in HT29 Colon Cancer Cells
Hojat Adeli, Changiz Ahmadizadeh, Mohammad Hossein Sadeghi Zali,
Volume 28, Issue 3 (6-2022)
Abstract
Aims Studies have shown that probiotic bacteria inhibit the onset and progression of carcinogenesis through different pathways. Our objective in this study was to determine the effect of probiotic bacteria on the expression of growth-related genes Rel A, IKB, and Casp3 in HT29 colon cancer cells
Methods & Materials In this study, the Lactobacillus brevis probiotic bacteria were first cultured, and after the supply of media conditioning, they were treated on HT29 cancer cells. The bacterium’s cytotoxic effect (bacterial T cells) was investigated using a microculture tetrazolium test (MTT). DNA was extracted from the treated cells, and a DNA Ladder assay was performed. Also, the 4′,6-diamidino-2-phenylindole (DAPI) test was performed to show cell apoptosis. After ribonucleic acid  (RNA) extraction and complementary DNA (cDNA) preparation to determine the mechanism of the effect of this bacterium on cellular signaling, the expression of growth-related genes Rel A, IKB, and Cas3 was measured using a real-time polymerase chain reaction (PCR) method.
Findings The microculture tetrazolium (MTT) test showed that L. b bacteria inhibit HT29 cells’ proliferation, induce apoptosis in these cells, and inhibit Rel A gene proliferation by increasing IKB gene expression. Also, 4′,6-diamidino-2-phenylindole (DAPI), and DNA ladder assay following the treatment of HT29 cells regarding the mentioned bacteria showed qualitative changes in cell apoptosis. In addition, real-time polymerase chain reaction (PCR) results showed that L. b increased Casp3 gene expression in HT29 colon cancer cells (P=0.038).
Conclusion Our findings indicate that L. b stimulates the apoptotic cell signaling pathway in HT29 colon cancer cells. It can be used as a new treatment strategy or therapy for colon cancer treatment.

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