Attenuated Salmonella carrying plasmid co-expressing HPV16 L1 and siRNA-E6 for cervical cancer therapy

Human papillomavirus (HPV) infection is the major etiological factor for cervical cancer. HPV prophylactic vaccines based on L1 virus-like particles have been considered as an effective prevention method. However, existing recombination vaccines are too expensive for developing countries. DNA vaccines might be a lower-cost and effective alternative. In this study, a plasmid (pcDNA3.1-HPV16-L1) and a co-expressing plasmid (pcDNA3.1-HPV16-L1-siE6) carried by attenuated Salmonella were constructed and their prevention and treatment effect on cervical cancer were observed, respectively. The results showed that pcDNA3.1-HPV16-L1 carried by attenuated Salmonella could induce the production of HPV16-L1 antibodies, IL-2 and INF-γ in mice serum, which presented its prevention effect on HPV. Subsequently, E6 and E7 gene silencing by pCG-siE6 inhibited the growth of cervical cancer both in vitro and in vivo. Furthermore, L1 up-regulation and E6/E7 down-regulation caused by co-expressing plasmid (pcDNA3.1-HPV16-L1-siE6) contributed to a significant anti-tumor effect on the mice. This study suggests that pcDNA3.1-HPV16-L1-siE6 carried by attenuated Salmonella has a synergistic effect of immune regulation and RNA interference in cervical cancer treatment.

Bioinformatics Prediction of miRNAs Targeting E6 and E7 Genes in Human Papillomavirus Types 16 and 18 in Cervical Cancer

Background and Aim: Cervical Cancer (CC) is the third most common malignancy in the women, the main cause of which is human papillomavirus (HPV). Both E6 and E7 oncogenes of the virus play an important role in its tumorigenesis. Today, methods available for screening CC are not capable of detecting the disease at an early stage. Therefore, it is important to identify new biomarkers for early detection of this cancer. For this purpose, in the present study, miRNAs targeting the two oncogenes E6 and E7 of human papillomavirus (types 16 and 18) were studied in CC by bioinformatics. Methods & Materials: First, using the NCBI database, the E6 and E7 gene sequences were obtained for both human papillomavirus types 16 and 18. Then, using the miRBase and RNA22 bioinformatics databases, the most appropriate targeting miRNAs for these genes were selected. Ethical Considerations: This study was approved by Ethics Committee of Arak University of Medical Sciences. Results: Based on the P obtained from bioinformatics databases, miRNA including miR-92a-5p (P=7.51e-2), miR-195-3p (P=2.24e-1), miR-34a-5p (P=2.73e-1) and miR-155-5p (P=4.95e-2) were introduced for the two genes E6 and E7. Conclusion: Results from bioinformatics studies revealed that of the four miRNAs identified, miR-155-5p and miR-92a-5p are probably the targeting miRNAs specific for the E6 and E7 genes, respectively. Therefore, it seems that these miRNAs can be a suitable candidate for in vitro studies in CC patients.

Possible Biomarkers and Therapeutic Targets for the Management of Cervical Cancer

Introduction: Cervical cancer is the most prevalent cancer in the world due to unusual extension of cervical cell. Cervical cancer occurs due to exposure of HPV (Human papillomavirus). According to WHO, it is the 4th most ordinary cancer in women. In 2018, approx 6.6% of population was affected around the world and 570,000 new cases were reported. In low and middle-income countries, 90% of cervical cancer deaths occur. Methods: Despite various factors that cause cervical cancer are included exposure to HPV, dysregulation of CASPASE enzyme, elevated expression of IAPs (Inhibitor apoptotic protein), E6 and E7 gene of HPV, inhibition of p53, BAK, p16 upregulation, CDK-inactivation causing cervical cancer, role of VEGF, role of estrogen and its receptor in cervical cancer. Results: Cervical cancer can be screened by Pep test. There are various therapies that can be used to treat cervical cancer. As these therapies have various side effects, so the world is moving to herbal formulations to treat cervical cancer. Conclusion: In this study, we will discuss cervical cancer, its cause, symptoms, pathophysiology and treatments. Early screening and detection can help in reducing the overall burden of cervical cancer in the near future.

Characteristics of mutant soybean lines at Satt100 and Satt319 loci linked with E7 gene

Aim. Аnalysis of genetic polymorphism of microsatellite loci Satt100 and Satt319 in 10 lines obtained by chemical mutagenesis, and 4 parental varieties – Oksana, Femida, Zolotysta, Podil’s’ka 416. Satt100 and Satt319 are flanking E7gene, which determines sensitivity of soybean to the length of the day in ripening phase. Methods. DNA were isolated from soybean seeds using the kit NeoPrep100 DNA. PCR were performed with microsatellites Satt100 and Satt319. The PCR products were fractionated in 7% polyacrylamide gels. Results. In the investigated lines were detected 5 alleles at the Satt100 locus and 3 alleles at the Satt319 locus. 42.9 % of the varieties and lines in this study were carriers of the «E» allele for Satt100. According to analysis of locus Satt319 64.2 % of varieties and lines were the carriers of «B» allele. Conclusions. Variety Oksana, mutant lines Oksana M №12, Femida M №29 – are carriers of the dominant allele E7. Variety Zolotysta and five mutant lines have recessive allele e7. In 75 % cases we have detected changes of alleles at microsatellite loci in mutant lines in comparison with parental forms. We assume that mutagenic reagents could affect at generative organs of soybean and lead totheir open flowering and crosspollinations. Keywords: Glycine max (L.), soybean, gene E7, microsatellite markers, photoperiodic sensitivity.