Journal of Cell and Molecular Research
Volume:12 Issue: 2, Winter and Spring 2021

Patients with ovarian cancer are mostly diagnosed at advanced stages which leads to poor prognosis and high mortality rate. Deregulation of lncRNA HOXD-AS1 expression associates with cancer development and metastasis. However, the expression level of this lncRNA in ovarian cancer is not determined.50paired ovarian tumors and their adjusted normal tissues were included in the study. Total RNA was extracted by TRIzol® Reagent and reverse-transcribed to cDNA using PrimeScript II cDNA synthesis kit. The expression levels of HOXD-AS1 were quantified by qRT-PCR and compared. The Roc curve analysis was used to evaluate the capacity of HOXD-AS1 as a biomarker for ovarian cancer. We observed that lncRNA HOXD-AS1 was significantly upregulated in ovarian tumors compared to their adjusted normal tissues (p <0.003). Moreover, the ROC curve analysis revealed that the lncRNA HOXD-AS1 expression level could discriminate tumoral and non-tumoral tissues with 85% sensitivity and 88% specificity. The lncRNA HOXD-AS1 expression level might be considered as a potential biomarker for ovarian cancer development.

Clostridium perfringens and novyi species are two important toxin-producing pathogens which pose a risk to the livestock health. Epsilon and alpha toxins are major toxins of these two pathogens, respectively. Advances in current vaccine industrialization lead to the utilization of toxin epitopes instead of the whole pathogen/toxoids to produce novel vaccines. In the present study, bioinformatics approaches were applied to design a fused protein containing both toxin fragments of interest with the highest antigenicity score for B-cells. To do so different specialized algorithms including I-TASSER, IEDB, ElliPro, PyDock and CLC Main Workbench were applied. The chimeric protein was successfully cloned, expressed, and purified using an immobilized-metal affinity chromatography for His-tagged proteins. During in vivo experiments on rabbits, the levels of immunization provided by the recombinant protein or native alpha and epsilon toxins were compared based on serological studies. Results indicated that the designed protein was able to stimulate effective immune responses against both alpha and epsilon toxins. This can be used as a proper strategy to design novel peptide-based subunit vaccines. Clostridium perfringens and novyi species are two important toxin-producing pathogens which pose a risk to the livestock health. Epsilon and alpha toxins are major toxins of these two pathogens, respectively. Advances in current vaccine industrialization lead to the utilization of toxin epitopes instead of the whole pathogen/toxoids to produce novel vaccines. In the present study, bioinformatics approaches were applied to design a fused protein containing both toxin fragments of interest with the highest antigenicity score for B-cells. To do so different specialized algorithms including I-TASSER, IEDB, ElliPro, PyDock and CLC Main Workbench were applied. The chimeric protein was successfully cloned, expressed, and purified using an immobilized-metal affinity chromatography for His-tagged proteins. During in vivo experiments on rabbits, the levels of immunization provided by the recombinant protein or native alpha and epsilon toxins were compared based on serological studies. Results indicated that the designed protein was able to stimulate effective immune responses against both alpha and epsilon toxins. This can be used as a proper strategy to design novel peptide-based subunit vaccines.

Small GTPases of RAS act as central regulators of intracellular signal transduction and translate external stimuli to the various cellular responses. Embryonic stem cell expressed RAS (ERAS) is a member of the RAS family that is specifically expressed in undifferentiated mouse embryonic stem cells, hepatic stellate cells and diverse human tumors, such as gastric, breast, brain, pancreatic, and colorectal tumors. Although ERAS belongs to GTPase family, it is an inefficient enzyme to hydrolyze GTP to GDP. Therefore, it remains mainly in its GTP-bound active form and contributes to sustained signal transduction. In comparison with classical members (HRAS, NRAS and KRAS4B), ERAS is known as a unique member, due to its temporal expression, remarkable amino acid sequence deviations and functional differences. Notably, ERAS has been recently proposed as a potential marker for drug resistance in several human tumors. In this minireview, I compare in great detail the biochemical properties of ERAS with conventional members of the RAS family, and discuss the main ERAS function in the control of the PI3K-AKT-mTORC survival pathway. Targeting this pathway may sensitize ERAS expressing cell populations to chemotherapy.

Acute myocardial infarction (MI) describes as an irreversible death of heart muscle which is initiated by a shortage of myocardium oxygen supply and accompanies by a complex of pro- and anti-inflammatory events. During the last decades, innate and adaptive immune responses are considered more serious for controlling myocardial infarction. As, it was confirmed that deregulated immune system which triggers excessive local and systemic inflammatory events is responsible for serious adverse effects associated with acute MI. Bone marrow activation, spleen monocytopoiesis, a remarkable increase of circulating cytokines and adhesion molecules, in addition to elevated levels of active peripheral leukocytes and platelets are playing significant roles in determining the clinical outcome of patients with MI. The previous experience demonstrated the failure of traditional harsh anti-inflammatory strategies. High mortality rate and poor quality of life observed for survivors of MI despite current progress in the field highlight the urgent need for such interdisciplinary studies in the context of molecular cardiology. Hence, unraveling the cellular and molecular events which are involved in the management of inflammatory responses post-MI is of special focus. The concept of immune regulation after myocardial infarction is not new, but our perception for dealing with the challenge has been changed during the last decades with gaining more in-depth molecular/immunological knowledge. It seems that fine-tuning the interplay between innate and adaptive immune responses and regulating their cross-talk should be in special focus to establish effective therapeutic strategies.

In this study, putative interactions between all of the retinoic acid (RA) ligands (all-trans (At), 9-cis (9c), and 13-cis (13c)), and VEGF receptors (VEGFR-1, -2 and -3) were investigated. It was performed considering the glycosylation status of the receptors to achieve a more reliable mode of interactions based on glycomics. We found that RAs may have a higher affinity for ligand-binding domains in VEGFRs. Furthermore, all RA isomers can strongly attach to VEGFR-3 receptor in comparison to other ones. It was also demonstrated that receptor dimerization of RAs may be less targeted. Moreover, regarding post-translational modifications, glycosylated structures showed conflicting binding energies. RAs may target the human vasculature, specifically lymph vessels, through VEGFR-3. In addition, the ligand binding-mediated activation of VEGFRs may be affected by these agents. Also, the glycosylation status of the receptors can interfere with these manners. Furthermore, our results confirmed that the consideration of carbohydrates in crystal structures is essential for a better interpretation of ligand/receptor interactions during drug discovery studies. Even though these observations improved our understanding of the binding patterns of RAs to VEGFRs, validation of these results needs further analysis to introduce these biomolecules as anti-VEGF remedies.

Proteomics is a powerful tool to identify effective proteins in the biochemical reactions of the insect body. Many proteins were reported in the gut lumen and tissues which are essential to complete the physiological role of the alimentary canal of Sunn pest. The gut microbiome of insects has a key role in the digestive process. In this study, for the first time, gut proteins of adult Sunn pest were extracted. These proteins were visualized and identified with two-dimensional polyacrylamide gel electrophoresis and mass spectrometry, respectively. Newly identified proteins include pyruvate dehydrogenase, oxidoreductase FAD-binding protein, hypothetical proteins, glycerol-3 phosphate dehydrogenase, conserved hypothetical proteins, ABC excinucleases, ABC-type transport systems and molecular chaperones. The accumulation of these metabolism proteins in the gut of Sunn pest indicates the importance of symbiotic proteins in the improvement of digestive activities and insect-bacteria interactions. Results suggest that the identified bacterial proteins can be considered as effective proteins in the process of nutrition and provide more gut-derived targets for enzyme engineering and development of biopesticides.

Real-time quantitative PCR (qRT-PCR) is often used as an effective experimental method for analyzing gene expression. In this method, normalization of target gene expression levels must be performed using housekeeping genes (HKGs). HKGs are used to compensate for difference between samples due to diverse quality and quality of RNAs and different reverse transcription yield. For an ideal reference gene, constant expression levels across different samples of one experiment is necessary. In the current study, expression stability of four candidate references genes including Beta actin (ACTB), glyceraldeyde-3-phosphate dehydrogenase (GAPDH), hypoxanthine guanine phosphoribosyl transferase (HPRT1) and Beta-2-Microglobulin (β2M) following retinoic acid (RA) treatment in embryonal carcinoma NCCIT cells were evaluated.NCCIT cells were exposed to RA (10 µM) for 14 days to induce differentiation. RT-qPCR for candidate references genes was performed and normalization between untreated and RA-treated cells was performed using identical sample input amounts. Expression of OCT4, SOX2, NANOG during RA-induced differentiation was assessed by quantitative real-time PCR. RT-qPCR results indicated significant difference in expression level of GAPDH between untreated (Ct mean: 19.36667± 0.28) and RA-treated (Ct mean: 28.94± 0.18) NCCIT cells. However, transcriptional level of ACTB, HPRT and β2M remained unchanged after RA treatment. qRT-PCR analysis using ACTB, HPRT and β2M showed treatment of NCCIT cells with RA lead to significant down regulation of OCT4 (79%), NANOG (71%) and SOX2 (96%) transcript. ACTB, HPRT and β2M were recognized as valid reference genes for analysis of gene expression during RA-induced differentiation of NCCIT cells, while GAPDH was not suitable.