Showing 3 results for Rt-Pcr
M Azarnoosh, S Zeinali, Sm Tabatabaei, A Ziaee,
Volume 12, Issue 3 (11-1998)
Abstract
Human growth hormone (hGH) genomic sequence containing 5 exons and 4
introns was cloned in pcDNA-3 and the constructed plasmid was subsequently used
for transfection ofNlli-3T3 cell line using lipofection technique. Expression of hGH
in stably transfected cells was assayed using ELISA. Total RNA was extracted from
transfected cells and hGH cDNA was amplified by RT-PCR using specific primers.
The product thus obtained was cloned in pGEM-T vector and the presence of the
growth hormone coding region was verified by restriction enzyme analysis and
Southern blotting. The hGH cDNA fragment was cloned in pQE-30 and the
expression of hGH gene in E. coli was assayed using ELISA and immunoblotting.
In this experiment 20.9 µg/mL purified rhGH was obtained.
Maryam Mansoori, Alireza Mirzaei, Isa Abdi Rad, Rahim Mahmodlou, Fatemeh Mansouri, Leili Saeednejad Zanjani, Zeynab Asadi- Lari, Zahra Madjd,
Volume 35, Issue 1 (1-2021)
Abstract
Background: GD2 synthase (GD2S) is the key enzyme required for ganglioside GD2 synthesis. It is commonly expressed in normal tissues and various cancers. Ganglioside GD2 is identified as a breast cancer stem cells (BCSCs) marker that promotes tumorigenesis. As GD2S has been found to be a useful molecular marker in neuroblastoma and retinoblastoma tumors, we suggest that it can be considered as a suitable candidate for the detection of CSCs in breast cancer tissues.
Methods: Expression of GD2S was examined in 65 breast tumors compared to adjacent normal tissues, applying quantitative real-time PCR (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA). The association between GD2S expression level and patients’ clinical characteristics was also assessed.
Results: Our findings showed that GD2S mRNA expression was significantly higher in breast cancer tissues in comparison to normal adjacent tissue samples (4.92-fold change, p<0.001) in advanced grades (p<0.001) and stages (p<0.001). It was also shown that GD2S protein expression was significantly higher in breast cancer tissues in comparison to normal adjacent tissues (4.86-fold change, p=0.010) in advanced grades (p=0.010), stages (p=0.005) and larger tumor size (p=0.002).
Conclusion: The current study showed that increased expression of GD2S in advanced breast cancer potentiates it as a promising tumor marker in these patients.
Samaneh Boroumand-Noughabi, Zahra Khoshnegah, Saeid Amel Jamehdar, Hossein Ayatollahi, Maryam Sheikhi, Mehrdad Rostami, Mohammad Reza Keramati,
Volume 36, Issue 1 (1-2022)
Abstract
Background: The autophagy machinery is reported to be employed by Coronaviruses during their replication. Beclin-1 (BECN1) and protein 1 light chain 3 (LC3) are two key elements in the autophagy process, and their inhibition can prevent the replication of some coronaviruses in vitro. Here, we aimed to investigate the expression levels of Beclin-1 and LC3 in COVID-19 patients and healthy controls, hoping to find new therapeutic targets.
Methods: This cross-sectional study was conducted in Imam Reza and Ghaem University Hospitals, Mashhad, Iran. Nasopharyngeal samples of 68 consecutive Covid-19 patients and 61 healthy controls, who have been referred to the laboratories for COVID-19 PCR testing between 21 March to 21 September 2021, were used in order to evaluate the expression of BECN1 and LC3 genes using the Real-time quantitative PCR method. Demographic and other laboratory findings of patients were extracted from the hospital electronic system. SPSS Statistics 16.0 and Graph Pad Prism 8.4.2 soft wares were used for statistical analysis. Non-parametric tests were used.
Results: BECN1 expression was significantly higher in COVID-19 patients compared to the controls (14.37±18.84 vs. 4.26±7.39, p=0.001). The expression of LC3 gene was significantly lower in patients compared to the controls (1.01±1.06 vs. 1.49±1.12, p=0.007). There was no significant correlation between the expression levels of BECN1 and LC3. Patients with lower BECN1 expression showed significantly higher RBC counts, higher Urea and lower HCO3 levels. The patients in LC3Low group showed significantly lower MCH, MCHC and PH levels compared to the others.
Conclusion: Regarding the significant difference in the expression of BECN1 and LC3 in COVID-19 patients compared to the controls, these molecules may have a role in the pathogenesis of this disease. In case of further confirmation of this role, these molecules may be used as possible therapeutic targets.
