Application of next generation sequencing technology in Mendelian movement disorders / 中南大学学报(医学版)

Next generation sequencing (NGS) has developed very rapidly in the last decade. Compared with Sanger sequencing, NGS has the advantages of high sensitivity and high throughput. Movement disorders are a common type of neurological disease. Although traditional linkage analysis has become a standard method to identify the pathogenic genes in diseases, it is getting difficult to find new pathogenic genes in rare Mendelian disorders, such as movement disorders, due to a lack of appropriate families with high penetrance or enough affected individuals. Thus, NGS is an ideal approach to identify the causal alleles for inherited disorders. NGS is used to identify genes in several diseases and new mutant sites in Mendelian movement disorders. This article reviewed the recent progress in NGS and the use of NGS in Mendelian movement disorders from genome sequencing and transcriptome sequencing. A perspective on how NGS could be employed in rare Mendelian disorders is also provided.

Advances in regulation of gene expression mediated by lncRNAs / 中南大学学报(医学版)

Long non-coding RNAs (lncRNAs) are a group of endogenous RNA molecules which exceed 200 nt in length, lack complete specific open reading frame, and completely lack or possessvery limited protein-coding capacity. Recent studies have revealed that lncRNAs participate in critical processes such as genomic imprinting, cell differentiation, and immune reaction, etc. lncRNAs regulate gene expression at the epigenetic, transcriptional and post-transcriptional levels by modulating chromatin remodeling and histone modifications, interfering the transcription, regulating patterns of alternative splicing, generating small RNAs, and modulating protein activation and localization. Through their numerous functions, lncRNAs play critical roles in the growth, development, senescence, death, and other important physiological and pathological processes. Further investigation into the regulation of gene expression mediated by lncRNAs will be of great value in the thorough understanding of pathogenies and provide new molecular markers and drug targets of diseases.

MicroRNAs and nonresolving inflammation-related cancer / 中南大学学报(医学版)

The link between nonresolving inflammation and cancer is well documented. On the one hand, epidemiologic evidence supports that approximately 25% of all human cancer worldwide is caused by nonresolving inflammation. On the other hand, inflammatory cells are found in the microenvironment of most, if not all, tumors. In the tumor micro-environment, inflammatory cells and molecules influence almost every aspect of cancer. MicroRNAs (miRNAs) participate in the initiation and progression of nonresolving inflammation-related cancer by regulating the key genes and related signaling pathways. Further investigation into the molecular mechanisms by which miRNAs carry out their functions will be of great value in the prevention, early diagnosis, and treatment of tumors.

Optimization of prokaryotic expression condition and purification of anti-cancer protein NOR1 in E.coli / 中南大学学报(医学版)

OBJECTIVE@#To optimize the induction condition of human NOR1 gene expression in E.coli. and purify NOR1 recombinant proteins.@*METHODS@#A full-length cDNA of human NOR1 was inserted into the corresponding region of pET28b expression vector to yield recombinant prokaryotic expression vector pET28b-NOR1. The prokaryotic expression vector pET28b-NOR1 was introduced into the bacterial host E.coli Rosettablue(DE3). Recombinant NOR1 protein was induced at different conditions. Induction condition was optimized to obtain high yield of recombinant protein. At last, the recombinant NOR1 protein was purified by Ni-IDE chromatography resin.@*RESULTS@#Recombinant NOR1 protein was induced by IPTG in a dose-dependent manner. Increase of kanamycin concentration and induction temperature resulted in high yield of recombinant protein. The most recombinant protein was found in inclusion bodies. The recombinant His-NOR1 protein was purified with Ni-IDE chromatography resin under denature condition.@*CONCLUSION@#IPTG, kanamycin concentration and temperature can affect the expression of recombinant NOR1 protein in pET28b system. High yield of recombinant NOR1 protein is achieved by inducing 1 mmol/L IPTG and 200 μg/mL kanamycin at 37 degree. Recombinant His-NOR1 protein with high purity is purified.

Transcriptomic regulation and molecular mechanism of polygenic tumor at different stages / 中南大学学报(医学版)

The research team on the National Key Scientific Program of China: "Transcriptomic regulation and molecular mechanism research of polygenic tumor at different stages" has focused on the field of transcriptomics of 4 common polygenic tumors, including nasopharyngeal carcinoma(NPC), breast cancer, colorectal cancer, and glioma. Extensive laboratory work has been carried out on the expression and regulation of tumor transcriptomics; identification of tumor suppressor/susceptible genes; mechanism of tumor epigenetics including miRNAs, and comparative study of specific gene/protein cluster of tumor transcriptomics and proteomics. Genes including SPLUNC1, LTF, BRD7, NOR1, BRCA1/2, PALB2, AF1Q, SOX17, NGX6, SOX7, and LRRC4 have been identified as the key transcriptional regulation genes during the stage of tumor initiation and invasion. Accordingly,the NPC gene signal regulation network of "SPLUNC1-miR-141-target genes", the breast cancer interaction signal pathway of "miR-193b-uPA",the glioma signal network of "miR-381- LRRC4-MEK/ERK/AKT", and the miRNA-target gene network of colorectal cancer metastasis related gene NGX6 have been thoroughly elucidated. These fruitful Results imply that the changes of key molecules in crucial signal pathway will cause severe dysfunction in signal transduction and gene regulation network in polygenic tumors, indicating that in the category of pathogenesis,these tumors may further classify as the "Disease of gene signal transduction and gene regulation network disorder". The researches have laid solid foundation for revealing the molecular mechanism and transcriptomic regulation of polygenic tumors at different stages.

The distinguishing of gastric cancer associated genes by digital gene expression displayer using serial analysis of gene expression database / 中国医师杂志

Objective To proceed deep data mining in large database obtained by Cancer Genome Anatomy Project and to distinguish the difference expression genes of gastric carcinoma.Methods The 300,783 serial analysis of gene expression records from 4 libraries of gastric cancer and 2 libraries of normal gastric tissue in database of the Cancer Genome Anatomy Project were analyzed by digital gene expression displayer.Results The 201 difference expression tags represented 136 genes(54 up-regulated and 82 down-regulated) and 65 ESTs(24 up-regulated and 41 down-regulated) were distinguished,and these genes were involved in biological process and signal transduction pathways,such as cell cycle regulation,cell proliferation and apoptosis.The gene expression profile in whole body normal and cancer tissues of PTMA were constructed by digital Northern based on serial analysis of gene expression database and by virtual Northern based on cDNA database,respectively.Conclusion The difference expression genes of gastric cancer can be distinguished effectively with bioinformatics,and the results will guide our further molecular biology research.If validated by molecular biology experiment,the difference expression genes will be used as molecular targets of gastric carcinoma.