Anti-myeloma STAT3 inhibitors and discovery strategies / 中国生化药物杂志

The STAT3 protein is a key transcription factor that delivers extracellular signals into nuclei in which it binds to a specific recognition element on DNA sequences and thus regulates the transcription of its target genes.STAT3 is almost expressed in all tissues and cells, therefore, it is extensively involved in both normal and aberrant cellular activities.For example, STAT3 plays an essential role in inflammatory reaction and tumorigenesis.STAT3 has been established as a therapeutic target of various cancers including multiple myeloma,a hematological malignancy from plasma cells.Some STAT3 inhibitors have been developed and extensively studied.In this article, we review the roles of STAT3 in multiple myeloma.The advances and strategies in the development of STAT3 inhibitors are also being discussed.

A cell-based high-throughput approach to identify inhibitors of influenza A virus

Influenza is one of the most common infections threatening public health worldwide and is caused by the influenza virus. Rapid emergence of drug resistance has led to an urgent need to develop new anti-influenza inhibitors. In this study we established a 293T cell line that constitutively synthesizes a virus-based negative strand RNA, which expresses Gaussia luciferase upon influenza A virus infection. Using this cell line, an assay was developed and optimized to search for inhibitors of influenza virus replication. Biochemical studies and statistical analyses presented herein demonstrate the sensitivity and reproducibility of the assay in a high-throughput format (Z' factor value>0.8). A pilot screening provides further evidence for validation of the assay. Taken together, this work provides a simple, convenient, and reliable HTS assay to identify compounds with anti-influenza activity.

Constructing a p53-fused dual luciferase reporter and verifying its function / 中南大学学报(医学版)

Objective:To construct a p53-fused dual luciferase reporter and to test whether this reporter can mimic wild-type p53 activities in a high-throughput screen.Methods:A restriction endonuclease site was added to each terminus and the stop codon of the wild-type full-length p53 open reading frame (ORF) was removed by PCR. A restriction endonuclease site was added to each terminus and the start codon of the ifrelfy luciferase ORF was removed by PCR. The two modified ORFs were inserted upstream of the IRES-induced renilla luciferase ORF in a CMV-derived vector. hTe p53 fusion protein was expressed in cells to test its MDM2-mediated degradation, subcellular localization, and induction of p53-responsive promoter. Results:hTe p53-fused dual luciferase reporter was successfully constructed. Atfer transfection into the host cells, the reporter expressing the p53 fusion protein that was degraded by oncoprotein MDM2, was mainly located inside the nucleus, and induced the p53-responsive promoter, respectively. Conclusion:hTe p53-fused dual luciferase reporter (p53FL/IRES/RL) can identify modulators of P53 protein level in a high-throughput screen of genetic or chemical libraries.

Technologies for Surface Display of Lipase / 中国生物工程杂志

Lipase is a kind of widely used hydrolase.Surface display is an efficient method of highthroughput screening for protein engineering of lipase.Besides,lipase displaying on surface of microorganism has many advantages,such as higher stability against high temperature and organic solvent,compared with free lipase,so the host strain displaying lipase can be used as wholecell biocatalyst,which has some advantages compared with traditional immobilization of lipase.There are three kinds of host strain for displaying lipase:phage,bacteria and yeast.The surface display of lipase in the three display systems were systematically discribed,and their current uses and possible trends in the future were discussed also.