yrammuS 要概87Sunchai Payungporn88Viroj BoonyaratanakornkitEnhanced Propagation Yield of Influenza Viruses for Vaccine Production through Cellular MicroRNAs Regulation(Project 2016)Production and Characterization of Novel Cell Penetrating Peptides for Targeted Therapy of Non-small Cell Lung Cancer(Project 2016)65Rep. Grant. Res., Asahi Glass Foundation (2018)Annual vaccination lessens the severity of clinical symptoms as well as dramatically reduces the risk of mortality from seasonal influenza viruses. Although chicken egg-based production is currently a major vaccine supply, this traditional system still faces various challenges, e.g., less flexible and scalable manufacturing, the possibility of an allergic effect, and changes in antigenicity. Therefore, cell culture-based system is approved as an alternative for seasonal influenza vaccine production. However, cellular microRNAs might play an important role in controlling the replication of viruses. The objective of this study is to improve the yield of seasonal influenza virus production via manipulation of the host microRNAs. Firstly, high-throughput microRNA profiles of Madin-Darby Canine Kidney (MDCK) cells upon infection with different subtypes/ lineages of seasonal influenza viruses were investigated using Next-Generation Sequencing (NGS) technology. After validated by reverse transcriptase polymerase chain reaction (qRT-PCR), the result revealed that four microRNAs, including cfa-miR-340, cfa-miR-146b, cfa-miR-197, and cfa-miR-215 were upregulated. Then, MDCK cells were transfected with the inhibitors of candidate microRNAs, followed by the infection of influenza viruses. The supernatants were collected at 48 hours post infection, and the amount of viruses was quantified by absolute quantitative qRT-PCR and haemagglutination (HA) method. The results demonstrated that cfa-miR-146b might be a candidate target of microRNA inhibitor in order to increase the production of pH1N1 and B/Yamagata viruses. Besides, cfa-miR-215 could be another good candidate for enhancing pH1N1 and B/Yamagata viral production. However, the microRNA inhibitor which targets cfa-miR-197 would be useful for the propagation of the H3N2 virus. According to our results, it might be inferred that microRNAs tend to target viral genes in a strain-specific manner, leading to suppressing viral replication. Conversely, propagation of influenza viruses could be enhanced by the utilization of microRNA inhibitors.Non-small cell lung cancer (NSCLC) constitutes the majority of all lung cancers. All currently available treatments have limited efficacy. Epidermal growth factor receptor or EGFR plays a critical role in the development and progression of NSCLC. High EGFR expression is associated with increased poor prognosis. Thus, EGFR served as an important therapeutic target for NSCLC. Previous studies demonstrated that progesterone receptor (PR) contains a polyproline domain (PPD) which directly interacts with proteins containing Src homology 3 (SH3) domains. Expression of PR-PPD inhibits NSCLC cell proliferation. In this study, we tested whether treatment of NSCLC with PR-PPD tagged with cecropin-B (CB) cell penetrating peptide (CB-PPD) inhibited EGFR signaling and NSCLC cell growth. Our results suggested that CB-PPD dose-dependently inhibited NSCLC cell viability. These data suggested that PR-PPD is the minimal domain sufficient in inhibiting NSCLC growth and could be used to further develop as novel NSCLC anticancer treatment in the future.
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