To study the effects of Shenqifuzheng injection (SFI) on micro-RNAs in human dendritic cells

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To study the effects of Shenqifuzheng injection (SFI) on micro-RNAs in human dendritic cells

 

Author: Yu, Wai Hong
Title: To study the effects of Shenqifuzheng injection (SFI) on micro-RNAs in human dendritic cells
Degree: D.H.Sc.
Year: 2014
Subject: Dendritic cells.
Medicine, Chinese.
Cancer -- Treatment
Hong Kong Polytechnic University -- Dissertations
Department: Faculty of Health and Social Sciences
Pages: 286 leaves : color illustrations ; 30 cm
Language: English
InnoPac Record: http://library.polyu.edu.hk/record=b2760484
URI: http://theses.lib.polyu.edu.hk/handle/200/7619
Abstract: Background: SHENQIFUZHENG Injection (SFI) has been used in China as a clinically-useful traditional Chinese medicine (TCM) in cancer management. Our study was to explore if SFI would enhance the immunomodulatory effects of the professional antigen presenting cells -- dendritic cells (DCs). DCs are significant in natural immunity against cancer. In this study, expression levels of HLA-DR and CD80 on DCs induced by SFI were measured by flow cytometric analysis. It had been reported that microRNA (miRNA) was central in the epigenetic control of gene expression at the cellular post-translational level. We hypothesized that by understanding the changes of miRNA profile expression during the treatment of DCs by the SFI, we might identify a few critical miRNA that would be instrumental in the DC activation and in turn leading to effective T cell priming. Method: This study used a 2-step approach. By first we were using a robust DC cell line to identify those miRNA that might be involved in DC activation; then followed by confirmation using monocyte-derived DC which was closer to real-life physiological contexts. THP-1 cell, a human monocytic leukaemia cell line, has previously been shown to be useful for the in vitro study of DC. In this study, we identified a number of potential miRNA candidates in the DCs activated by SFI using MicroRNA Array of 381 known human miRNAs (Sanger miRBase, version 14). miR-22, miR-107 and miR-200a were then selected from the over 30 potential miRNA candidates with relative expression fold change >2 indentified in accordance to the computational evidences retrieved in the annotation database pipeline miRBase. These three miRNAs were further examined for any change in expression in a confirmatory study using the monocyte-derived DCs (MDDC) from peripheral blood mononuclear cells of healthy normal subjects (n=7) through the quantification by Real Time RT-PCR quantification. These results were correlated with the expression levels of HLA-DR and CD80 on the MDDCs using flow cytometry.
Results: The confirmatory results did not show any consistent upregulation in the expressions of miR22, miR107 and miR-200a due to SFI across the seven MDDC samples. In addition, test results on MDDCs upon treatment of SFI in two different dilutions of 1/20 and 1/40 respectively for 24 hours incubation also demonstrated insignificant expression changes of HLA-DR and CD80 (p>0.05). Furthermore, the correlation of the expression of the three mi-RNAs and the surface expression levels of HLA-DR and CD80 were not statistically significant for the MDDCs with treatment of SFI diluted in 1/20 for 1 hour, SFI diluted in 1/20 for 4 hours, and SFI diluted in 1/40 for 1 hour (p>0.05). Conclusions: Increased expression of miRNA of miR-22, miR-107 and miR-200a were found only in the cell line of THP-1, but failed to be shown consistently in the MDDCs from seven human peripheral blood samples. Thus whether these three mi-RNAs were really important epigenetic regulators during the process of SFI on DC remain unknown; and will require more future investigations by using multiple authentic DC sources.

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