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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 is correlated with CRC.(A) Convergent primers can amplify circular RNAs and linear RNAs. Divergent primers amplify circular RNAs only in cDNA compared with genomic DNA (gDNA). GAPDH is linear control. (B) The cir-ITCH was expressed at a higher level in approximately 75.6% of the CRC adjacent tissues compared to match CRC tissues. The expression level of cir-ITCH was analyzed by qRT-PCR based on Taq-man and normalized to GAPDH. Data are represented as mean±SEM from three independent experiments. (C) Random primers and oligodT primers were used respectively in the reverse transcription experiments. The predicted cir-ITCH is absent in poly (A) enriched samples. (D) The predicted cir-ITCH is react against to RNase R treatment. 2-tailed student’s t-test were used in test the differences between groups *p < 0.05 compared to control.
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pone.0131225.g001: cir-ITCH is correlated with CRC.(A) Convergent primers can amplify circular RNAs and linear RNAs. Divergent primers amplify circular RNAs only in cDNA compared with genomic DNA (gDNA). GAPDH is linear control. (B) The cir-ITCH was expressed at a higher level in approximately 75.6% of the CRC adjacent tissues compared to match CRC tissues. The expression level of cir-ITCH was analyzed by qRT-PCR based on Taq-man and normalized to GAPDH. Data are represented as mean±SEM from three independent experiments. (C) Random primers and oligodT primers were used respectively in the reverse transcription experiments. The predicted cir-ITCH is absent in poly (A) enriched samples. (D) The predicted cir-ITCH is react against to RNase R treatment. 2-tailed student’s t-test were used in test the differences between groups *p < 0.05 compared to control.

Mentions: Two sets of ITCH primers were designed for this study: a divergent set to amplify only the circular form and a convergent set to amplify the linear forms. We confirmed that the circular form was amplified using the divergent primers. cDNA and genomic DNA were used as the controls. As expected, no amplification was observed using the divergent primers on genomic DNA. GAPDH was used as the linear control (Fig 1A).


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 is correlated with CRC.(A) Convergent primers can amplify circular RNAs and linear RNAs. Divergent primers amplify circular RNAs only in cDNA compared with genomic DNA (gDNA). GAPDH is linear control. (B) The cir-ITCH was expressed at a higher level in approximately 75.6% of the CRC adjacent tissues compared to match CRC tissues. The expression level of cir-ITCH was analyzed by qRT-PCR based on Taq-man and normalized to GAPDH. Data are represented as mean±SEM from three independent experiments. (C) Random primers and oligodT primers were used respectively in the reverse transcription experiments. The predicted cir-ITCH is absent in poly (A) enriched samples. (D) The predicted cir-ITCH is react against to RNase R treatment. 2-tailed student’s t-test were used in test the differences between groups *p < 0.05 compared to control.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0131225.g001: cir-ITCH is correlated with CRC.(A) Convergent primers can amplify circular RNAs and linear RNAs. Divergent primers amplify circular RNAs only in cDNA compared with genomic DNA (gDNA). GAPDH is linear control. (B) The cir-ITCH was expressed at a higher level in approximately 75.6% of the CRC adjacent tissues compared to match CRC tissues. The expression level of cir-ITCH was analyzed by qRT-PCR based on Taq-man and normalized to GAPDH. Data are represented as mean±SEM from three independent experiments. (C) Random primers and oligodT primers were used respectively in the reverse transcription experiments. The predicted cir-ITCH is absent in poly (A) enriched samples. (D) The predicted cir-ITCH is react against to RNase R treatment. 2-tailed student’s t-test were used in test the differences between groups *p < 0.05 compared to control.
Mentions: Two sets of ITCH primers were designed for this study: a divergent set to amplify only the circular form and a convergent set to amplify the linear forms. We confirmed that the circular form was amplified using the divergent primers. cDNA and genomic DNA were used as the controls. As expected, no amplification was observed using the divergent primers on genomic DNA. GAPDH was used as the linear control (Fig 1A).

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