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Downregulation of microRNA-409-3p promotes aggressiveness and metastasis in colorectal cancer: an indication for personalized medicine.

Liu M, Xu A, Yuan X, Zhang Q, Fang T, Wang W, Li C - J Transl Med (2015)

Bottom Line: Aberrant miR-409-3p expression has been reported in several cancers.However, the clinical significance and functions of miR-409-3p in human CRC were not entirely clear. miR-409-3p expression levels were determined in 45 pairs of primary CRC and their corresponding adjacent non-tumor tissues by qPCR.Notably, we found the NLK could be a potential target of miR-409-3p.

View Article: PubMed Central - PubMed

Affiliation: Department of Gastrointestinal Surgery, The First Affiliated Hospital of Bengbu Medical College, Bengbu, 233030, China. mulinliu11@163.com.

ABSTRACT

Background: MicroRNAs play an ess ential role in colorectal cancer development and progression. Aberrant miR-409-3p expression has been reported in several cancers. However, the clinical significance and functions of miR-409-3p in human CRC were not entirely clear.

Methods: miR-409-3p expression levels were determined in 45 pairs of primary CRC and their corresponding adjacent non-tumor tissues by qPCR. The effects of ectopic expression of miR-409-3p on CRC cells proliferation, wound healing, metastasis were investigated by CCK-8, transwell assay and peritoneal spreading nude mice model.

Results: Statistical analysis of clinical cases revealed that low miR-409-3p expression had inclinations towards lager tumor size and local invasion. Ectopic expression of miRNA mimics suggested that miR-409-3p could inhibits the abilities of proliferation, wound healing, metastasis and invasion in CRC cells. Notably, we found the NLK could be a potential target of miR-409-3p.

Conclusion: Our results suggest that miR-409-3p functions as a tumor suppressor by inhibiting the development and metastasis of CRC, suggesting that miR-409-3p is expected to become a new diagnostic marker and a new target of the treatment of CRC.

No MeSH data available.


Related in: MedlinePlus

miR-409-3p targets the 3′-UTR of the oncogene NLK.a The putative binding sites of miR-409-3p in NLK 3′-UTR region were predicted. The matched seed sequences were indicated by vertical lines. b The minimum free energy (mfe) required for RNA hybridization was predicted by RNAhybrid software (mfe: −9.3 kcal/mol). c Schematic graph of the putative binding sites of miR-409-3p in the NLK 3′UTR and the mutation in miR-409-3p-binding sites. d miR-409-3p mimics down-regulated luciferase activities controlled by wild-type NLK 3′UTR (*P < 0.01), but did not affect luciferase activity controlled by mutant NLK 3′UTR. The results are means of three independent experiments ± SD.
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Fig7: miR-409-3p targets the 3′-UTR of the oncogene NLK.a The putative binding sites of miR-409-3p in NLK 3′-UTR region were predicted. The matched seed sequences were indicated by vertical lines. b The minimum free energy (mfe) required for RNA hybridization was predicted by RNAhybrid software (mfe: −9.3 kcal/mol). c Schematic graph of the putative binding sites of miR-409-3p in the NLK 3′UTR and the mutation in miR-409-3p-binding sites. d miR-409-3p mimics down-regulated luciferase activities controlled by wild-type NLK 3′UTR (*P < 0.01), but did not affect luciferase activity controlled by mutant NLK 3′UTR. The results are means of three independent experiments ± SD.

Mentions: To identify how miR-409-3p functions in CRC cells, computational prediction of miR-409-3p targets was performed. We searched miR-409-3p’s putative targets using online search tools (e.g. TargetScan, Microrna.org and RNAhybrid) [13–17]. The genes predicted by all the used programs were considered candidate targets of miR-409-3p. Among all the hits, NLK caused our attention (Figure 7a, b). We then performed luciferase reporter assays to verify a direct interaction between miR-409-3p and the 3′UTR of NLK. Luciferase reporters were constructed containing either a wild-type NLK 3′UTR sequence containing the miR-409-3p binding site (NLK-3′UTRwt), or a mutated 3′UTR (NLK-3′UTRmut) (Figure 7c). The relative luciferase activity of the NLK-3′UTRwt reporter was markedly suppressed by miR-409-3p mimics compared with that of NLK-3′UTRmut in a miR-409-3p dependent manner (Figure 7d). This result strongly indicates that 3′UTR of NLK carries the direct binding seed of miR-409-3p.Figure 7


Downregulation of microRNA-409-3p promotes aggressiveness and metastasis in colorectal cancer: an indication for personalized medicine.

Liu M, Xu A, Yuan X, Zhang Q, Fang T, Wang W, Li C - J Transl Med (2015)

miR-409-3p targets the 3′-UTR of the oncogene NLK.a The putative binding sites of miR-409-3p in NLK 3′-UTR region were predicted. The matched seed sequences were indicated by vertical lines. b The minimum free energy (mfe) required for RNA hybridization was predicted by RNAhybrid software (mfe: −9.3 kcal/mol). c Schematic graph of the putative binding sites of miR-409-3p in the NLK 3′UTR and the mutation in miR-409-3p-binding sites. d miR-409-3p mimics down-regulated luciferase activities controlled by wild-type NLK 3′UTR (*P < 0.01), but did not affect luciferase activity controlled by mutant NLK 3′UTR. The results are means of three independent experiments ± SD.
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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Fig7: miR-409-3p targets the 3′-UTR of the oncogene NLK.a The putative binding sites of miR-409-3p in NLK 3′-UTR region were predicted. The matched seed sequences were indicated by vertical lines. b The minimum free energy (mfe) required for RNA hybridization was predicted by RNAhybrid software (mfe: −9.3 kcal/mol). c Schematic graph of the putative binding sites of miR-409-3p in the NLK 3′UTR and the mutation in miR-409-3p-binding sites. d miR-409-3p mimics down-regulated luciferase activities controlled by wild-type NLK 3′UTR (*P < 0.01), but did not affect luciferase activity controlled by mutant NLK 3′UTR. The results are means of three independent experiments ± SD.
Mentions: To identify how miR-409-3p functions in CRC cells, computational prediction of miR-409-3p targets was performed. We searched miR-409-3p’s putative targets using online search tools (e.g. TargetScan, Microrna.org and RNAhybrid) [13–17]. The genes predicted by all the used programs were considered candidate targets of miR-409-3p. Among all the hits, NLK caused our attention (Figure 7a, b). We then performed luciferase reporter assays to verify a direct interaction between miR-409-3p and the 3′UTR of NLK. Luciferase reporters were constructed containing either a wild-type NLK 3′UTR sequence containing the miR-409-3p binding site (NLK-3′UTRwt), or a mutated 3′UTR (NLK-3′UTRmut) (Figure 7c). The relative luciferase activity of the NLK-3′UTRwt reporter was markedly suppressed by miR-409-3p mimics compared with that of NLK-3′UTRmut in a miR-409-3p dependent manner (Figure 7d). This result strongly indicates that 3′UTR of NLK carries the direct binding seed of miR-409-3p.Figure 7

Bottom Line: Aberrant miR-409-3p expression has been reported in several cancers.However, the clinical significance and functions of miR-409-3p in human CRC were not entirely clear. miR-409-3p expression levels were determined in 45 pairs of primary CRC and their corresponding adjacent non-tumor tissues by qPCR.Notably, we found the NLK could be a potential target of miR-409-3p.

View Article: PubMed Central - PubMed

Affiliation: Department of Gastrointestinal Surgery, The First Affiliated Hospital of Bengbu Medical College, Bengbu, 233030, China. mulinliu11@163.com.

ABSTRACT

Background: MicroRNAs play an ess ential role in colorectal cancer development and progression. Aberrant miR-409-3p expression has been reported in several cancers. However, the clinical significance and functions of miR-409-3p in human CRC were not entirely clear.

Methods: miR-409-3p expression levels were determined in 45 pairs of primary CRC and their corresponding adjacent non-tumor tissues by qPCR. The effects of ectopic expression of miR-409-3p on CRC cells proliferation, wound healing, metastasis were investigated by CCK-8, transwell assay and peritoneal spreading nude mice model.

Results: Statistical analysis of clinical cases revealed that low miR-409-3p expression had inclinations towards lager tumor size and local invasion. Ectopic expression of miRNA mimics suggested that miR-409-3p could inhibits the abilities of proliferation, wound healing, metastasis and invasion in CRC cells. Notably, we found the NLK could be a potential target of miR-409-3p.

Conclusion: Our results suggest that miR-409-3p functions as a tumor suppressor by inhibiting the development and metastasis of CRC, suggesting that miR-409-3p is expected to become a new diagnostic marker and a new target of the treatment of CRC.

No MeSH data available.


Related in: MedlinePlus