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Circulating plant miRNAs can regulate human gene expression in vitro

View Article: PubMed Central - PubMed

ABSTRACT

While Brassica oleracea vegetables have been linked to cancer prevention, the exact mechanism remains unknown. Regulation of gene expression by cross-species microRNAs has been previously reported; however, its link to cancer suppression remains unexplored. In this study we address both issues. We confirm plant microRNAs in human blood in a large nutrigenomics study cohort and in a randomized dose-controlled trial, finding a significant positive correlation between the daily amount of broccoli consumed and the amount of microRNA in the blood. We also demonstrate that Brassica microRNAs regulate expression of human genes and proteins in vitro, and that microRNAs cooperate with other Brassica-specific compounds in a possible cancer-preventive mechanism. Combined, we provide strong evidence and a possible multimodal mechanism for broccoli in cancer prevention.

No MeSH data available.


Related in: MedlinePlus

Effect of miR160 transfection on targets expression.Ratio of expression of tested genes after transfection of miR160 in two NSCLC cell lines compared to expression in negative control (AllStars, scrambled sequence). siPORT = transfection reagent. NT = not treated. p-values (t-test): *0.05–0.01, **0.01–0.001.
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f3: Effect of miR160 transfection on targets expression.Ratio of expression of tested genes after transfection of miR160 in two NSCLC cell lines compared to expression in negative control (AllStars, scrambled sequence). siPORT = transfection reagent. NT = not treated. p-values (t-test): *0.05–0.01, **0.01–0.001.

Mentions: We have selected two NSCLC cell lines with the highest expression of at least one of the genes identified as putative broccoli miRNA targets. We then transfected the two cancer cell lines with miR160 mimic, and measured the potential downregulation of the targets. Figure 3 shows that 8 out of 9 tested genes were significantly downregulated by miR160 transfection ((p < 0.05); MCM5 was not changed). Supplementary Figure 5 shows the amount of transfected miR160 per cell line, while Supplementary Table 6 lists p-values for comparisons of all samples against AllStars Negative Control. There was no significant difference between AllStars Negative Control, transfection reagent and not treated cells (Supplementary Table 6). NCBP2 and SHMT2 were significantly reduced both as transcript and protein, while MTHFD2 was reduced as transcript but not protein, after miR160 transfection (Supplementary Figure 6). Human microRNA regulation of CDC6 has already been described as a mechanism of prevention of lung cancer formation20, while PYCR1 and MTHFD2 have been shown to be required for breast tumorigenesis in vitro21.


Circulating plant miRNAs can regulate human gene expression in vitro
Effect of miR160 transfection on targets expression.Ratio of expression of tested genes after transfection of miR160 in two NSCLC cell lines compared to expression in negative control (AllStars, scrambled sequence). siPORT = transfection reagent. NT = not treated. p-values (t-test): *0.05–0.01, **0.01–0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5015063&req=5

f3: Effect of miR160 transfection on targets expression.Ratio of expression of tested genes after transfection of miR160 in two NSCLC cell lines compared to expression in negative control (AllStars, scrambled sequence). siPORT = transfection reagent. NT = not treated. p-values (t-test): *0.05–0.01, **0.01–0.001.
Mentions: We have selected two NSCLC cell lines with the highest expression of at least one of the genes identified as putative broccoli miRNA targets. We then transfected the two cancer cell lines with miR160 mimic, and measured the potential downregulation of the targets. Figure 3 shows that 8 out of 9 tested genes were significantly downregulated by miR160 transfection ((p < 0.05); MCM5 was not changed). Supplementary Figure 5 shows the amount of transfected miR160 per cell line, while Supplementary Table 6 lists p-values for comparisons of all samples against AllStars Negative Control. There was no significant difference between AllStars Negative Control, transfection reagent and not treated cells (Supplementary Table 6). NCBP2 and SHMT2 were significantly reduced both as transcript and protein, while MTHFD2 was reduced as transcript but not protein, after miR160 transfection (Supplementary Figure 6). Human microRNA regulation of CDC6 has already been described as a mechanism of prevention of lung cancer formation20, while PYCR1 and MTHFD2 have been shown to be required for breast tumorigenesis in vitro21.

View Article: PubMed Central - PubMed

ABSTRACT

While Brassica oleracea vegetables have been linked to cancer prevention, the exact mechanism remains unknown. Regulation of gene expression by cross-species microRNAs has been previously reported; however, its link to cancer suppression remains unexplored. In this study we address both issues. We confirm plant microRNAs in human blood in a large nutrigenomics study cohort and in a randomized dose-controlled trial, finding a significant positive correlation between the daily amount of broccoli consumed and the amount of microRNA in the blood. We also demonstrate that Brassica microRNAs regulate expression of human genes and proteins in vitro, and that microRNAs cooperate with other Brassica-specific compounds in a possible cancer-preventive mechanism. Combined, we provide strong evidence and a possible multimodal mechanism for broccoli in cancer prevention.

No MeSH data available.


Related in: MedlinePlus