No abstract

Expression of bacterial luciferase enzyme (lux) in eukaryotic cells would provide a new bioreporter system for in vivo imaging and diagnostics technology. In spite of this, until now only a few efforts have been made to express bacterial luciferase enzyme in eukaryotic cells. We attempted to synthesize an expression construct of luxA and luxB genes from Vibrio fischeri. The luxA and luxB genes were cloned into the MCS of pTZ57R via the 5' kpnI, BamHI and BamHI, EcoRI restriction sites to generate pTZ57R/luxA and pTZ57R/luxB respectively, then newly synthesized constructs were cleaved with the same enzymes and respectively cloned into the pcDNA3.1(+) (Hyg) and pcDNA3.1(+) (Neo) expression vectors to create pcDNA3.1(+) (Hyg)/luxA and pcDNA3.1(+) (neo)/luxB. Recombinant constructs were cotransfected to the NIH3T3 cell line. Gene expression was confirmed by reverse transcription-polymerase chain reaction, sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting; in addition, bioluminescence characteristics of transfected NIH3T3 cell lines were evaluated by decanal supplement. In conclusion, in the current research, separate vector systems were constructed, which are composed of bacterial luciferase genes (luxA and luxB) that accordingly have not already been reported. These results hold promise toward the potential development of an autonomous light-generating lux reporter system in eukaryotic cells.

Background: Breast cancer (BC) is one of the most common cancers among women and is the main cause of cancer-related mortalities in the female population. The main cause of BC is not fully understood yet; however, many genes have been identified as risk factors that increase susceptibility to this disease. Objectives: The aim of this study was to evaluate the expression of ABCC12 gene in patients with ductal breast carcinoma and its relationship with other biomarkers including estrogen receptor (ER), progesterone receptor (RP) and human epidermal growth factor receptor 2 (HER-2). Methods: This study included nine women diagnosed with ductal breast carcinoma as cases and five healthy women as controls. RNA extraction from breast tissue and cDNA synthesis were performed, and the expression of ABCC12 gene was evaluated using the real time polymerase chain reaction (PCR) method. After preparing formalin-fixed paraffin-embedded breast tissues, immunohistochemical evaluation of these samples and chromogenic in situ hybridization (CISH) of ER, PR and HER-2 were performed. Results: The obtained results showed that the expression of ABCC12 gene in all the breast cancer tissues was 3.74 times higher than in controls (P = 0.007). Also, the expression of ABCC12 gene increased by 3.94 times in luminal B (P = 0.002), 3.54 times in HER-2 (1+) (P = 0.0006), 3.86 times in ER (+) (P = 0.002) and 3.63 times in PR (+) (P = 0.0005). Conclusions: Our study showed that the evaluation of the ABCC12 gene expression as a one of multi-drug resistance protein members can be used as a prognostic factor to identify the stages of BC and its therapeutic approaches.

Introduction: Several genetic loci are suspected to be involved in prostate cancer, including the hereditary prostate cancer. RNASEL regulates cell proliferation and apoptosis through the IFN-regulated 2-5A pathway that mediates anti proliferative activities and has been suggested to be a candidate tumor suppressor gene. There are numerous nucleotide variants identified in the RNASEL gene. The two most commonly found variants in the U.S. non-Hispanic Caucasian population are the non-synonymous variants: 1385 G>A; Arg462Gln; and 1623T>G; Asp 541 Glu has been reported. The Arg 462 Gln variant reduces the ability of the cell to cause apoptosis in response to activation by 2-5 (A) and also has three times less enzymatic activity than normal, whereas the Asp 541 Glu variant has no known effect on RNASEL protein function. There continues to be much debate over whether these common variants increase the risk of prostate cancer. It has been also reported that the Arg 462 Gln AA genotype associated with both increased prostate cancer in U.S. Caucasian sample groups and decreased prostate cancer risk in Caucasian and Japanese sample groups Moreover, the Asp 541 Glu variant within RNASEL has been reported that the GG and TT genotypes were associated with an increased risk for prostate cancer in Japanese and European-American samples, respectively. On the other hand, a significant negative association of the TT genotype with prostate cancer in Swedish Caucasian samples has been reported .However; the role of these common mutations is debatable. The aim of this study was to investigate the role of common variants of RNASE L gene; Arg 462 Gln and Asp 541 Glu in Iranian men with prostate cancer. Patients and Methods:Blocks of paraffin-embedded tissue from 46 patients with adenoma carcinoma prostate cancer and 50 patients with hyperplasia prostate tissue and also fifty healthy blood controls with PSA<4 was enrolled in this current research study. ARMS-PCR amplification was performed with specific primers. Results and Conclusions: This is the first study that investigates the association of prostate cancer risk with RNASEL variants in Iranian prostate cancer patients. Our result indicated that the frequency of the AA, GG and AG genotypes in the RNASEL 462 SNP in Tumors were 30%, 13.3% and 56.66% respectively, in hyperplasia was 30.3% ,15.15% and 54.54% respectively and in controls was 26%, 14% and 60% respectively .The frequency of the TT, GG and TG genotypes in the RNASEL 541 SNP in Tumors were 3%, 0 % and 93.47% respectively, in hyperplasia were 10 %,0% and 90% and in blood controls were 8%, 0% and 92% respectively. Our finding indicated that these variations were not a significant association with prostate cancer risk in Iranian patients and the other variations of this gene maybe involve in Iranian prostate cancer patients. Note: This abstract was not presented at the meeting. Citation Format: Sirweh Molla, Massoud Ghaffarpour, Somayeh Saeeda, Seyed Werya Hossieni, Sina Mirzaahmadi, Mohammad Reza Rahmany. Evaluation of the frequency of RNASE L in 462 and 541 variants in Iranian prostate cancer patients. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1848. doi:10.1158/1538-7445.AM2014-1848

Background: Infertility is considered as a common problem appears in about 10-12% of couples in their reproductive ages. Ring finger protein 38 (RNF38) gene is a ubiquitinprotein ligase that can regulate Protein 53 (P53) and affect cellular motility. Objective: Considering the role of P53 on cellular motility and RNF38 on the regulation of P53, the present study aimed to assess the difference between RNF38 and P53 genes expression in normozoospermic and asthenospermic samples as a diagnostic biomarker in males. Materials and Methods: The present study was conducted among 21 asthenospermics and 63 healthy individuals. First, the real-time polymerase chain reaction technique was applied to measure the expression level of the P53 and RNF38 genes extracted from sperm samples, and the glyceraldehyde-3phosphate dehydrogenase gene was selected as the reference gene. Results: An increase and a decrease occurred in the level of P53 and RNF38 genes expressions in asthenospermic and normozoospermic samples, respectively. In addition, a significant difference was observed between increasing P53 gene expression (p < 0.001), reducing RNF38 one, and decreasing sperm motility (p < 0.001) in asthenospermic cells compared to that of normozoospermic ones. Conclusion: Based on the results, an increase in the expression of the P53 gene and a decrease in the expression of the RNF38 gene had a significant relationship with asthenospermia in men. Therefore, it is expected that an effective step should be adopted to diagnose the asthenospermia expression pattern by using these results. Key words: RNF38, P53, Asthenozoospermia, Motility.