New Molecular Markers for Prostate Cancer Diagnosis.

PURPOSE: Prostate cancer (PCa) is the second most commonly diagnosed cancer and the sixth leading cause of cancer death among men worldwide. Biomarkers are an important tool in the early detection of PCa. Prostate-specific antigen (PSA) is one of the oldest biomarkers for the early detection of PCa. Digital rectal exam (DRE) is another screening test for PCa detection, which is considered as an irritating experience for patients. Biopsy is still the most reliable method for PCa diagnosis; however, patients are prone to complications. Therefore, developing non-invasive and accurate methods for PCa screening seems urgent to avoid unnecessary biopsies. There has been remarkable development in PCa molecular biomarkers discovery, largely through progress in omics technologies. Due to the many benefits of liquid biopsies, a significant set of PCa diagnostic kits have been developed using urine samples. Despite the unique benefits of these kits, there are still many challenges to their widespread use in clinics. Here, we have reviewed the latest developments of PCa biomarkers in liquid biopsies. METHODS: Literature on biomarkers for diagnosis of PCa was reviewed during the past two decades. RESULTS: PSA, PHI, PCA3, and 4K score are among the commonly used markers for PCa diagnosis which have been used over a long-moderate length of time with multiple studies on their performance. We performed a review of their performance. Newer markers are among RNA and DNA markers. Multiple non-coding RNAs (mi-RNAs) were reviewed and their performance on Pca diagnosis was reviewed. Long noncoding RNAs (Lnc RNAs) including PlncRNA-1, HOTAIR, SchLAP-1, MALAT1, MEG3, and PRCAT17.3 were summarized. mRNA markers including TMPRSS2:ERG, and HOXC6 were presented. DNA-based markers including PTEN, HOXB13, and BRCA2 were reviewed. Finally, the use of CircRNAs was reviewed for PCa diagnosis. CONCLUSION: Many reviewed RNA-based biomarkers have promising results in the diagnosis of PCa.

In vitro Inhibition of Border Disease Virus Replication With Lentivirus-Mediated shRNAs.

Background: Border disease is believed to be one of the most important diseases in the animal husbandry industry, which has not yet been eradicated in Iran. The development of approaches based on the application of interfering RNA (RNAi) for antiviral therapy has attracted a great deal of attention over the recent years. The present research was conducted to design, construct, and apply shRNA against the NS3 gene of BDV to evaluate the prevention of BDV proliferation in the cell culture system. For this purpose, the suitable oligonucleotide sequence of NS3 gene coding was selected utilizing BDV- X818 strain. Afterwards, using shRNA design software, shRNA molecules were designed and synthesized. These shRNAs were cloned into the desired vectors and were finally transfected in HEK293T cells employing the third generation of lentiviral packaging system. Subsequently, these shRNA expressing lentiviruses were transduced to the MDBK cell line to challenge to border virus. In order to evaluate the efficacy of shRNAs, the viral infectious titer and RNA copy number were calculated with TCID50 and Real-time RT-PCR tests, respectively. Results: The results revealed that shRNAs 1, 2, and 3 decreased viral RNA by more than 90% compared to the control groups. BDV titer noticeably decreased after the challenge with shRNAs 1, 2, and 3 from ~88% up to 99% in comparison with the control groups. Conclusions: Overall, it could be concluded that RNAi may be considered as a strong treatment proposal against viruses, such as BDV.

Lentivirus expressing shRNAs inhibit the replication of contagious ecthyma virus by targeting DNA polymerase gene.

BACKGROUND: Contagious ecthyma or Orf is known as a zoonotic disease remains prevalently worldwide despite the application of some control strategies against it. RNAi particularly shRNA provides us with the chance to tackle this obstacle by an encouraging new approach. The current study indicates the design and experiment of third-generation lentivirus packaging systems delivering shRNAs to inhibit Orf virus (ORFV) replication and infection. Given the importance of DNA-pol gene in virus replication, in this study, three shRNAs against this gene were designed and cloned into lentiviral vectors to stabilize the expression of shRNAs. After producing lentivectors expressing ORFV-DNA- pol in HEK293T cells, the synthesized shRNAs were applied to downregulate viral replication and gene expression. The reduction in viral titer and RNA was evaluated by TCID50 test as well as real-time RT-PCR. The results were then analyzed in comparison with the control group. RESULTS: Designed shRNAs significantly reduced virus yield approximately 90 to 97% and 96.8 to 99.4%, respectively compared to the control groups (cells infected with ORFV and infected with ORFV and scrambled vector) by TCID50 test. Real-time RT-PCR revealed a dramatic reduction in the expression of viral RNA approximately 99% compared to cells infected with ORFV and from 92.6 to 99%, respectively compared to cells infected with ORFV and scrambled vector. CONCLUSIONS: Therefore, it can be stated that RNAi is capable of being used as a potent therapeutically option against viruses like ORFV.