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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

cir-ITCH involves in the regulation Wnt/b-catenin signaling pathway in vivo.(A) The linear correlations between the cir-ITCH expression levels and linear ITCH were tested. The relative expression value was normalized by GAPDH expression level. (B) A TCF luciferase reporter assay was performed. The luciferase activity was normalized to the Renilla luciferase activity. (C) The protein levels of Wnt3a and β-catenin was assessed in CRC cells (HCT116 cells and SW480 cells) by Western blot. (D) The mRNA level of c-myc and cyclinD1was detected by quantitative RT-PCR after transfected with cir-ITCH or Control cells in CRC cells. (the upper is c-myc and the lower is cyclinD1) Data are mean±SEM and representative of three independent experiments. (E) HCT116 and SW480 cells were seeded in 96-well plates after been transfected, and cell proliferation was performed daily for 3 days using the CCK-8 assay. Six replicates for each group and the experiment repeated three times. Data are mean±SEM. *P<0.05 compared with controls.
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pone.0131225.g003: cir-ITCH involves in the regulation Wnt/b-catenin signaling pathway in vivo.(A) The linear correlations between the cir-ITCH expression levels and linear ITCH were tested. The relative expression value was normalized by GAPDH expression level. (B) A TCF luciferase reporter assay was performed. The luciferase activity was normalized to the Renilla luciferase activity. (C) The protein levels of Wnt3a and β-catenin was assessed in CRC cells (HCT116 cells and SW480 cells) by Western blot. (D) The mRNA level of c-myc and cyclinD1was detected by quantitative RT-PCR after transfected with cir-ITCH or Control cells in CRC cells. (the upper is c-myc and the lower is cyclinD1) Data are mean±SEM and representative of three independent experiments. (E) HCT116 and SW480 cells were seeded in 96-well plates after been transfected, and cell proliferation was performed daily for 3 days using the CCK-8 assay. Six replicates for each group and the experiment repeated three times. Data are mean±SEM. *P<0.05 compared with controls.

Mentions: Recently, cir-ITCH was reported to regulate gene expression through its role as an miRNA sponge, thus protecting the miRNAs’ target genes. Therefore, we tested the correlation between cir-ITCH and ITCH in an additional 15 pairs of CRC adjuvant non-cancerous tissues. The results showed that patients with higher cir-ITCH expression levels in the CRC tissues had a substantial up-regulation of linear ITCH (R2 = 0.32, P < 0.01; Fig 3A).


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)

cir-ITCH involves in the regulation Wnt/b-catenin signaling pathway in vivo.(A) The linear correlations between the cir-ITCH expression levels and linear ITCH were tested. The relative expression value was normalized by GAPDH expression level. (B) A TCF luciferase reporter assay was performed. The luciferase activity was normalized to the Renilla luciferase activity. (C) The protein levels of Wnt3a and β-catenin was assessed in CRC cells (HCT116 cells and SW480 cells) by Western blot. (D) The mRNA level of c-myc and cyclinD1was detected by quantitative RT-PCR after transfected with cir-ITCH or Control cells in CRC cells. (the upper is c-myc and the lower is cyclinD1) Data are mean±SEM and representative of three independent experiments. (E) HCT116 and SW480 cells were seeded in 96-well plates after been transfected, and cell proliferation was performed daily for 3 days using the CCK-8 assay. Six replicates for each group and the experiment repeated three times. Data are mean±SEM. *P<0.05 compared with controls.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4482251&req=5

pone.0131225.g003: cir-ITCH involves in the regulation Wnt/b-catenin signaling pathway in vivo.(A) The linear correlations between the cir-ITCH expression levels and linear ITCH were tested. The relative expression value was normalized by GAPDH expression level. (B) A TCF luciferase reporter assay was performed. The luciferase activity was normalized to the Renilla luciferase activity. (C) The protein levels of Wnt3a and β-catenin was assessed in CRC cells (HCT116 cells and SW480 cells) by Western blot. (D) The mRNA level of c-myc and cyclinD1was detected by quantitative RT-PCR after transfected with cir-ITCH or Control cells in CRC cells. (the upper is c-myc and the lower is cyclinD1) Data are mean±SEM and representative of three independent experiments. (E) HCT116 and SW480 cells were seeded in 96-well plates after been transfected, and cell proliferation was performed daily for 3 days using the CCK-8 assay. Six replicates for each group and the experiment repeated three times. Data are mean±SEM. *P<0.05 compared with controls.
Mentions: Recently, cir-ITCH was reported to regulate gene expression through its role as an miRNA sponge, thus protecting the miRNAs’ target genes. Therefore, we tested the correlation between cir-ITCH and ITCH in an additional 15 pairs of CRC adjuvant non-cancerous tissues. The results showed that patients with higher cir-ITCH expression levels in the CRC tissues had a substantial up-regulation of linear ITCH (R2 = 0.32, P < 0.01; Fig 3A).

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