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miR-34a screened by miRNA profiling negatively regulates Wnt/β-catenin signaling pathway in Aflatoxin B1 induced hepatotoxicity.

Zhu L, Gao J, Huang K, Luo Y, Zhang B, Xu W - Sci Rep (2015)

Bottom Line: Here, we applied Illumina deep sequencing technology for high-throughout profiling of microRNAs in HepG2 cells lines after treatment with AFB1.Anti-miR-34a can significantly relieved the down-regulated β-catenin and its downstream genes, c-myc and Cyclin D1, and the S-phase arrest in cell cycle induced by AFB1 can also be relieved.These results suggested that AFB1 might down-regulate Wnt/β-catenin signaling pathway in HepG2 cells by up-regulating miR-34a, which may involve in the mechanism of liver tumorigenesis.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Food Safety, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, China, 100083.

ABSTRACT
Aflatoxin-B1 (AFB1), a hepatocarcinogenic mycotoxin, was demonstrated to induce the high rate of hepatocellular carcinoma (HCC). MicroRNAs (miRNAs) participate in the regulation of several biological processes in HCC. However, the function of miRNAs in AFB1-induced HCC has received a little attention. Here, we applied Illumina deep sequencing technology for high-throughout profiling of microRNAs in HepG2 cells lines after treatment with AFB1. Analysis of the differential expression profile of miRNAs in two libraries, we identified 9 known miRNAs and 1 novel miRNA which exhibited abnormal expression. KEGG analysis indicated that predicted target genes of differentially expressed miRNAs are involved in cancer-related pathways. Down-regulated of Drosha, DGCR8 and Dicer 1 indicated an impairment of miRNA biogenesis in response to AFB1. miR-34a was up-regulated significantly, down-regulating the expression of Wnt/β-catenin signaling pathway by target gene β-catenin. Anti-miR-34a can significantly relieved the down-regulated β-catenin and its downstream genes, c-myc and Cyclin D1, and the S-phase arrest in cell cycle induced by AFB1 can also be relieved. These results suggested that AFB1 might down-regulate Wnt/β-catenin signaling pathway in HepG2 cells by up-regulating miR-34a, which may involve in the mechanism of liver tumorigenesis.

No MeSH data available.


Related in: MedlinePlus

qRT-PCR analyses of mRNA expression levels.Expression levels were normalized using β-actin. Data are presented as the means ± SD. Three independent experiments are conducted in HepG2 cells. *p < 0.05, **p < 0.01.
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f7: qRT-PCR analyses of mRNA expression levels.Expression levels were normalized using β-actin. Data are presented as the means ± SD. Three independent experiments are conducted in HepG2 cells. *p < 0.05, **p < 0.01.

Mentions: The targets were predicted according to the method above by different miRNA target prediction algorithms, which can be helpful in minimizing the number of putative or false positive targets. The mRNA expression of AGO/IGF1/mTOR (miR-99a), E2F3/Cyclin D1(miR-16), PTEN (miR-19a/b), Cyclin E/β-catenin/Bcl2 (miR-34a) were strongly correlated with its corresponding miRNAs shown in the parentheses. The mRNAs were decreased significantly after AFB1 treatment. All the primers used in the RT-PCR analyses were listed in supplementary Table 1B. The expression tendency of these mRNA targets was opposite to the expression of their corresponding miRNAs, as shown in Fig. 7.


miR-34a screened by miRNA profiling negatively regulates Wnt/β-catenin signaling pathway in Aflatoxin B1 induced hepatotoxicity.

Zhu L, Gao J, Huang K, Luo Y, Zhang B, Xu W - Sci Rep (2015)

qRT-PCR analyses of mRNA expression levels.Expression levels were normalized using β-actin. Data are presented as the means ± SD. Three independent experiments are conducted in HepG2 cells. *p < 0.05, **p < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4645126&req=5

f7: qRT-PCR analyses of mRNA expression levels.Expression levels were normalized using β-actin. Data are presented as the means ± SD. Three independent experiments are conducted in HepG2 cells. *p < 0.05, **p < 0.01.
Mentions: The targets were predicted according to the method above by different miRNA target prediction algorithms, which can be helpful in minimizing the number of putative or false positive targets. The mRNA expression of AGO/IGF1/mTOR (miR-99a), E2F3/Cyclin D1(miR-16), PTEN (miR-19a/b), Cyclin E/β-catenin/Bcl2 (miR-34a) were strongly correlated with its corresponding miRNAs shown in the parentheses. The mRNAs were decreased significantly after AFB1 treatment. All the primers used in the RT-PCR analyses were listed in supplementary Table 1B. The expression tendency of these mRNA targets was opposite to the expression of their corresponding miRNAs, as shown in Fig. 7.

Bottom Line: Here, we applied Illumina deep sequencing technology for high-throughout profiling of microRNAs in HepG2 cells lines after treatment with AFB1.Anti-miR-34a can significantly relieved the down-regulated β-catenin and its downstream genes, c-myc and Cyclin D1, and the S-phase arrest in cell cycle induced by AFB1 can also be relieved.These results suggested that AFB1 might down-regulate Wnt/β-catenin signaling pathway in HepG2 cells by up-regulating miR-34a, which may involve in the mechanism of liver tumorigenesis.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Food Safety, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, China, 100083.

ABSTRACT
Aflatoxin-B1 (AFB1), a hepatocarcinogenic mycotoxin, was demonstrated to induce the high rate of hepatocellular carcinoma (HCC). MicroRNAs (miRNAs) participate in the regulation of several biological processes in HCC. However, the function of miRNAs in AFB1-induced HCC has received a little attention. Here, we applied Illumina deep sequencing technology for high-throughout profiling of microRNAs in HepG2 cells lines after treatment with AFB1. Analysis of the differential expression profile of miRNAs in two libraries, we identified 9 known miRNAs and 1 novel miRNA which exhibited abnormal expression. KEGG analysis indicated that predicted target genes of differentially expressed miRNAs are involved in cancer-related pathways. Down-regulated of Drosha, DGCR8 and Dicer 1 indicated an impairment of miRNA biogenesis in response to AFB1. miR-34a was up-regulated significantly, down-regulating the expression of Wnt/β-catenin signaling pathway by target gene β-catenin. Anti-miR-34a can significantly relieved the down-regulated β-catenin and its downstream genes, c-myc and Cyclin D1, and the S-phase arrest in cell cycle induced by AFB1 can also be relieved. These results suggested that AFB1 might down-regulate Wnt/β-catenin signaling pathway in HepG2 cells by up-regulating miR-34a, which may involve in the mechanism of liver tumorigenesis.

No MeSH data available.


Related in: MedlinePlus