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. 2015 Dec 29;6(42):44758-80.
doi: 10.18632/oncotarget.5815.

"V体育ios版" Radiation and SN38 treatments modulate the expression of microRNAs, cytokines and chemokines in colon cancer cells in a p53-directed manner

Surajit Pathak  1 , Wen-Jian Meng  1   2 , Suman Kumar Nandy  3 , Jie Ping  1 , Atil Bisgin  1 , Linda Helmfors  4 , Patrik Waldmann  5 , Xiao-Feng Sun  1
Affiliations

Radiation and SN38 treatments modulate the expression of microRNAs, cytokines and chemokines in colon cancer cells in a p53-directed manner

Surajit Pathak et al. Oncotarget. .

VSports手机版 - Abstract

Aberrant expression of miRNAs, cytokines and chemokines are involved in pathogenesis of colon cancer. However, the expression of p53 mediated miRNAs, cyto- and chemokines after radiation and SN38 treatment in colon cancer remains elusive. Here, human colon cancer cells, HCT116 with wild-type, heterozygous and a functionally null p53, were treated by radiation and SN38. The expression of 384 miRNAs was determined by using the TaqMan® miRNA array, and the expression of cyto- and chemokines was analyzed by Meso-Scale-Discovery instrument. Up- or down-regulations of miRNAs after radiation and SN38 treatments were largely dependent on p53 status of the cells VSports手机版. Cytokines, IL-6, TNF-α, IL-1β, Il-4, IL-10, VEGF, and chemokines, IL-8, MIP-1α were increased, and IFN-γ expression was decreased after radiation, whereas, IL-6, IFN-γ, TNF-α, IL-1β, Il-4, IL-10, IL-8 were decreased, and VEGF and MIP-1α were increased after SN38 treatment. Bioinformatic analysis pointed out that the highly up-regulated miRNAs, let-7f-5p, miR-455-3p, miR-98, miR-155-5p and the down-regulated miRNAs, miR-1, miR-127-5p, miR-142-5p, miR-202-5p were associated with colon cancer pathways and correlated with cyto- or chemokine expression. These miRNAs have the potential for use in colon cancer therapy as they are related to p53, pro- or anti-inflammatory cyto- or chemokines after the radiation and SN38 treatment. .

Keywords: chemokines; colon cancer cells; cytokines; miRNAs; p53 V体育安卓版. .

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Conflict of interest statement

COMPETING INTEREST

The authors declare that they have no competing interests.

"VSports app下载" Figures

Figure 1
Figure 1. Distribution of the density of raw Ct values for the individual samples without normalization
The expression levels of 379 miRNAs and 5 control assays were analyzed in HCT116p53+/+, HCT116p53+/− and HCT116p53−/−. A. is for the expression of miRNA without or with the radiation treatment. B. is for the expression of miRNA without or with the SN38 treatment. The both A and B illustrate that the different cell lines had almost the same trends of raw Ct values.
Figure 1
Figure 1. Distribution of the density of raw Ct values for the individual samples without normalization
The expression levels of 379 miRNAs and 5 control assays were analyzed in HCT116p53+/+, HCT116p53+/− and HCT116p53−/−. A. is for the expression of miRNA without or with the radiation treatment. B. is for the expression of miRNA without or with the SN38 treatment. The both A and B illustrate that the different cell lines had almost the same trends of raw Ct values.
Figure 2
Figure 2. Heatmaps for the expression of miRNAs in HCT116p53+/+, HCT116p53+/− and HCT116p53−/− cell lines with the radiation and SN38 treatment, the cell lines grouped in vertical columns and miRNAs arranged horizontally by the similarity of the expression to one another
The clusters were based on the Euclidean distance between log2 (T/N) values. T/N= Treatment/Normal, and > 0.5 of miRNA expression was considered as up-regulation. A. is for the expression of miRNA after the radiation treatment, and B. is for the expression of miRNAs after SN38 treatment. The more white color shows the more up-regulation, and the more green color is for the more down-regulation. Here only those miRNAs with log2 (T/N) values from all the three cell lines were considered in the heatmap. HCT116p53+/+ cells had more up-regulated miRNAs and HCT116p53−/− cells had more down-regulated miRNAs of the each treatment.
Figure 3
Figure 3. Venn diagram showing up-regulated A. and down-regulated B. miRNAs after radiation and SN38 treatment in HCT116p53+/+, HCT116p53+/− and HCT116p53−/− cell lines
The Empirical Bayes mixture method implemented in the R-package EBarrays was used for analyzing differential expression (DE) of miRNAs in the cells. The numbers of overlapping significant miRNAs between the cell lines were visualized with Venn-diagrams.
Figure 4
Figure 4. Colorectal cancer pathway with interaction between significant up-regulated miRNAs, let-7f-5p, miR-455-3p, miR-98, miR-155-5p, and down-regulated miRNAs, miR-1, miR-127-5p, miR-142-5p, miR-202-5p after radiation and SN38 treatments and their target genes
Figure 5
Figure 5. Heatmap of four most significant up-regulated miRNA, let-7f-5p, miR-455-3p, miR-98 and miR-155-5p, as well as four most down-regulated miRNA, miR-1, miR-127-5p, miR-142-5p and miR-202-5p after radiation and SN38 treatments, and different pathways by clustering from pathway union
Each colorful square means that pathway contains target genes of the miRNA. The more red color shows the more significant relationship between the miRNA and the pathway.
Figure 6
Figure 6. The expression of cytokine and chemokine in HCT116p53+/+, HCT116p53+/− and HCT116p53−/− cell lines after radiation A. and SN38 treatments B
Cytokines, IL-6, TNF-α, IL-1β, Il-4, IL-10, VEGF, chemokines, IL-8, MIP-1α were increased, and IFN-γ expression was decreased after 48 hr of 2GY radiation, whereas, IL-6, IFN-γ, TNF-α, IL-1β, Il-4, IL-10, IL-8 expression were decreased, and VEGF MIP-1α were increased after SN38 treatment. For all of the experimental samples and all of the cyto- and chemokines a z-score was calculated as z=X−μσ. If z-score was greater than 1.96 then it up-regulated and down-regulated if the z-score was less than 1.96, corresponding to a p-value of 0.05.
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