Limits...
cir-ITCH plays an inhibitory role in colorectal cancer by regulating the Wnt/β-catenin pathway.

Huang G, Zhu H, Shi Y, Wu W, Cai H, Chen X - PLoS ONE (2015)

Bottom Line: We found that cir-ITCH expression was typically down-regulated in CRC compared to the peritumoral tissue.This result, as well as several follow-up experiments, showed that cir-ITCH could increase the level of ITCH, which is involved in the inhibition of the Wnt/β-catenin pathway.Therefore, our results showed that cir-ITCH plays a role in CRC by regulating the Wnt/β-catenin pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgical Oncology, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, P.R. China.

ABSTRACT
Noncoding RNAs (ncRNAs) are the dominant product of eukaryotic transcription. These products range from short microRNAs (miRNAs) to long intergenic noncoding RNAs (lincRNAs). Circular RNAs composed of exonic sequences represent an understudied form of ncRNA that was discovered more than 20 years ago. Using a TaqMan-based reverse transcriptase polymerase chain reaction assay, we analyzed the relationship between cir-ITCH expression and colorectal cancer (CRC) in a total of 45 CRCs and paired adjacent non-tumor tissue samples. We found that cir-ITCH expression was typically down-regulated in CRC compared to the peritumoral tissue. This result, as well as several follow-up experiments, showed that cir-ITCH could increase the level of ITCH, which is involved in the inhibition of the Wnt/β-catenin pathway. Therefore, our results showed that cir-ITCH plays a role in CRC by regulating the Wnt/β-catenin pathway.

No MeSH data available.


Related in: MedlinePlus

The sponges role of cir-ITCH in CRC cells.(A) Relative luciferase activity of the psiCHECK-2-ITCH constructs co-transfected with miR-20a, miR-7, miR-214 and inhibitor in HCT116 and SW480 cells. In cir-ITCH hyper-expression cells, there were no significant differences in luciferase activity when psiCHECK-2-ITCH-binding site with miRNAs were cotransfected into HCT116 and SW480 cells. Six replicates for each group and the experiment repeated at least three times. Data are mean±SEM. (B) HCT116 and SW480 cells after transfected with cir-ITCH and Control cells were respectively transfected with miR-20a, miR-7 and inhibitor for 24 h and were then further exposed to actinomycin D for 1, 2 and 3 h. Cells were harvested and the stability of cir-ITCH mRNA was analyzed by qRT-PCR relative to time 0 after blocking new RNA synthesis with actinomycin D; data are mean±SEM, normalized to GAPDH.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4482251&req=5

pone.0131225.g002: The sponges role of cir-ITCH in CRC cells.(A) Relative luciferase activity of the psiCHECK-2-ITCH constructs co-transfected with miR-20a, miR-7, miR-214 and inhibitor in HCT116 and SW480 cells. In cir-ITCH hyper-expression cells, there were no significant differences in luciferase activity when psiCHECK-2-ITCH-binding site with miRNAs were cotransfected into HCT116 and SW480 cells. Six replicates for each group and the experiment repeated at least three times. Data are mean±SEM. (B) HCT116 and SW480 cells after transfected with cir-ITCH and Control cells were respectively transfected with miR-20a, miR-7 and inhibitor for 24 h and were then further exposed to actinomycin D for 1, 2 and 3 h. Cells were harvested and the stability of cir-ITCH mRNA was analyzed by qRT-PCR relative to time 0 after blocking new RNA synthesis with actinomycin D; data are mean±SEM, normalized to GAPDH.

Mentions: We performed the same experiment in cir-ITCH hyper-expression cells. The results showed that there were no significant differences in luciferase activity when the psiCHECK-2-ITCH-binding site with miR-7 and miR-20a was transfected into HCT116 and SW480 cells but that there was a significant difference for miR-214 (1 pmol miRNA-7: 1.147 ± 0.041 versus 1.236 ± 0.042, P = 0.069; 40 pmol miRNA-7: 1.138 ± 0.015 versus 1.236 ± 0.042, P = 0.12; 1 pmol miRNA-20a: 2.424 ± 0.017 versus 2.526 ± 0.058, P = 0.11; 40 pmol miRNA-20a: 2.345 ± 0.04 versus 2.526 ± 0.058, P = 0.069; 1 pmol miRNA-214: 1.539 ± 0.038 versus 1.877 ± 0.039, P = 0.001; 40 pmol miRNA-214: 1.407 ± 0.051 versus 1.877 ± 0.039, P = 0.004) (Fig 2A).


cir-ITCH plays an inhibitory role in colorectal cancer by regulating the Wnt/β-catenin pathway.

Huang G, Zhu H, Shi Y, Wu W, Cai H, Chen X - PLoS ONE (2015)

The sponges role of cir-ITCH in CRC cells.(A) Relative luciferase activity of the psiCHECK-2-ITCH constructs co-transfected with miR-20a, miR-7, miR-214 and inhibitor in HCT116 and SW480 cells. In cir-ITCH hyper-expression cells, there were no significant differences in luciferase activity when psiCHECK-2-ITCH-binding site with miRNAs were cotransfected into HCT116 and SW480 cells. Six replicates for each group and the experiment repeated at least three times. Data are mean±SEM. (B) HCT116 and SW480 cells after transfected with cir-ITCH and Control cells were respectively transfected with miR-20a, miR-7 and inhibitor for 24 h and were then further exposed to actinomycin D for 1, 2 and 3 h. Cells were harvested and the stability of cir-ITCH mRNA was analyzed by qRT-PCR relative to time 0 after blocking new RNA synthesis with actinomycin D; data are mean±SEM, normalized to GAPDH.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4482251&req=5

pone.0131225.g002: The sponges role of cir-ITCH in CRC cells.(A) Relative luciferase activity of the psiCHECK-2-ITCH constructs co-transfected with miR-20a, miR-7, miR-214 and inhibitor in HCT116 and SW480 cells. In cir-ITCH hyper-expression cells, there were no significant differences in luciferase activity when psiCHECK-2-ITCH-binding site with miRNAs were cotransfected into HCT116 and SW480 cells. Six replicates for each group and the experiment repeated at least three times. Data are mean±SEM. (B) HCT116 and SW480 cells after transfected with cir-ITCH and Control cells were respectively transfected with miR-20a, miR-7 and inhibitor for 24 h and were then further exposed to actinomycin D for 1, 2 and 3 h. Cells were harvested and the stability of cir-ITCH mRNA was analyzed by qRT-PCR relative to time 0 after blocking new RNA synthesis with actinomycin D; data are mean±SEM, normalized to GAPDH.
Mentions: We performed the same experiment in cir-ITCH hyper-expression cells. The results showed that there were no significant differences in luciferase activity when the psiCHECK-2-ITCH-binding site with miR-7 and miR-20a was transfected into HCT116 and SW480 cells but that there was a significant difference for miR-214 (1 pmol miRNA-7: 1.147 ± 0.041 versus 1.236 ± 0.042, P = 0.069; 40 pmol miRNA-7: 1.138 ± 0.015 versus 1.236 ± 0.042, P = 0.12; 1 pmol miRNA-20a: 2.424 ± 0.017 versus 2.526 ± 0.058, P = 0.11; 40 pmol miRNA-20a: 2.345 ± 0.04 versus 2.526 ± 0.058, P = 0.069; 1 pmol miRNA-214: 1.539 ± 0.038 versus 1.877 ± 0.039, P = 0.001; 40 pmol miRNA-214: 1.407 ± 0.051 versus 1.877 ± 0.039, P = 0.004) (Fig 2A).

Bottom Line: We found that cir-ITCH expression was typically down-regulated in CRC compared to the peritumoral tissue.This result, as well as several follow-up experiments, showed that cir-ITCH could increase the level of ITCH, which is involved in the inhibition of the Wnt/β-catenin pathway.Therefore, our results showed that cir-ITCH plays a role in CRC by regulating the Wnt/β-catenin pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgical Oncology, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, P.R. China.

ABSTRACT
Noncoding RNAs (ncRNAs) are the dominant product of eukaryotic transcription. These products range from short microRNAs (miRNAs) to long intergenic noncoding RNAs (lincRNAs). Circular RNAs composed of exonic sequences represent an understudied form of ncRNA that was discovered more than 20 years ago. Using a TaqMan-based reverse transcriptase polymerase chain reaction assay, we analyzed the relationship between cir-ITCH expression and colorectal cancer (CRC) in a total of 45 CRCs and paired adjacent non-tumor tissue samples. We found that cir-ITCH expression was typically down-regulated in CRC compared to the peritumoral tissue. This result, as well as several follow-up experiments, showed that cir-ITCH could increase the level of ITCH, which is involved in the inhibition of the Wnt/β-catenin pathway. Therefore, our results showed that cir-ITCH plays a role in CRC by regulating the Wnt/β-catenin pathway.

No MeSH data available.


Related in: MedlinePlus