Screening of Nucleostemin p53-Independent Signal Pathway in NB4 Leukemia Cell

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Screening of Nucleostemin p53-Independent Signal Pathway in NB4 Leukemia Cell

Journal Title: International Journal of Contemporary Medical Research - Year 2017, Vol 4, Issue 4

Abstract

Introduction: Nucleostemin is a newly discovered nucleoprotein which is found to be highly expressed in the undifferentiated cells like stem cells and cancer cells. It is an important G1/S checkpoint regulator. The aim of study was to find the specific mechanism of nucleostemin p-53 independent signal pathway. Finding NS p53- independent signal pathway can be an important tool use in gene therapy of cancer patients. Material and Methods: The present study comprised of sample size of 2719 differentially expressed genes from NB4 experimental group, 847 genes were up-regulated and 1872 genes were down-regulated. RNA quantity and quality were measured by NanoDrop ND-1000. For DNA microarray, the Whole Human Genome Oligo Microarray was used and sequences were compiled from a broad source survey, and then verified and optimized by alignment to the assembled human genome. Agilent Feature Extraction software (version 11.0.1.1) was used to analyze the acquired array images. Quantile normalization and subsequent data processing were performed with using the GeneSpring GX v12.0 software package (AgilentTechnologies). Pathway analysis and GO analysis were applied to determine the roles of these differentially expressed genes played in these biological pathways or GO terms. Finally, Hierarchical Clustering was performed to show the distinguishable gene expression profiling among samples. Results: Inhibition of Nucleostemin mRNA expression succeeded in human leukemia cell lines NB4 by using lentiviral technology, the rate of Inhibition in NB4 cells was 58%, Real-time PCR demonstrated the reliability of the data. These differentially expressed genes may involve in great kinds of functions and signal pathways through GO analysis and Pathway analysis, in which the association with MAPK signaling pathway and NODlike receptor signaling pathway for up-regulated the most related and significant. RNA transport and mRNA surveillance pathway is the most significant and related pathways for down-regulated. RNA transport from the nucleus to the cytoplasm is fundamental for gene expression. Conclusion: The data reveals that MAPK signaling pathway is one of the most effected pathway for upregulated genes, thus it is one of the important pathways, that is p53 independent and modifying it can cause NB4 cell apoptosis. Similarly, RNA transport signaling pathway as a most effected pathway of downregulated genes.

Authors and Affiliations

Abdallah Dlykan, Sun Xiaoli, Atul Dwivedi, Wang Ningning

Keywords

Nucleostemin; Leukemia Cell; Lentivirus Vector; DNA Microarray; RNA Transport; MAPK Signaling Pathway; Cell Apoptosis

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