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MicroRNA‑205 promotes the tumorigenesis of nasopharyngeal carcinoma through targeting tumor protein p53-inducible nuclear protein 1.

Nie G, Duan H, Li X, Yu Z, Luo L, Lu R, Ji Z, Zhang W - Mol Med Rep (2015)

Bottom Line: The present study aimed to explore the potential role of miR‑205 in NPC.The expression of miR‑205 was increased in NPC tissues compared with that in normal tissues.Overexpression of miR‑205 was found to promote the proliferation, migration and invasion of NPC‑derived cells, while apoptosis was suppressed.

View Article: PubMed Central - PubMed

Affiliation: Department of Otolaryngology, Peking University Shenzhen Hospital, Shenzhen, Guangdong 518036, P.R. China.

ABSTRACT
Nasopharyngeal carcinoma (NPC) is a common type of cancer in southern China, miRNAs have been shown to be involved in the tumorigenesis of multiple cancer types. The present study aimed to explore the potential role of miR‑205 in NPC. Reverse transcription quantitative polymerase chain reaction was used to determine the expression levels of miR‑205 in 20 fresh NPC specimens and 20 normal nasopharyngeal tissues. The function of miR‑205 in the proliferation, migration, invasion and apoptosis of NPC‑derived cells was detected by MTT assay, colony formation assay, wound healing assay, Transwell assay and flow cytometry. Furthermore, a target gene of miR‑205 was identified using the luciferase reporter assay. The expression of miR‑205 was increased in NPC tissues compared with that in normal tissues. Overexpression of miR‑205 was found to promote the proliferation, migration and invasion of NPC‑derived cells, while apoptosis was suppressed. Tumor protein p53-inducible nuclear protein 1 was identified as a target gene of miR‑205. Overall, the present study demonstrated that miR‑205 may function as an oncogene in NPC tumorigenesis.

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TP53IPN1 is a target gene of miR-205. (A) Fragments of TP53INP1 3′UTR, which contained the WT potential binding sites was synthesized and the MU fragments were generated by replacing G with T and A with C on the putative binding sites. (B) The target gene was verified by luciferase reporter assay. Co-transfection with miR-205 and TP53INP1-3′UTR significantly decreased the luciferase activity by 50.9% (P=0.006), 36.4% (P=0.03) and 43.9% (P=0.006) in CNE2, 6-10B and 9-4E cells, respectively. However, the activity of the reporter containing the mutated seed sequence was not obviously changed. Values are expressed as the mean ± standard deviation. *P<0.05 vs. NC. LUC, luciferase; WT, wild-type; Mut; mutant; UTR, untranslated region; miR, microRNA; NC, negative control; TP53INP1, tumor protein p53-inducible nuclear protein 1.
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f6-mmr-12-04-5715: TP53IPN1 is a target gene of miR-205. (A) Fragments of TP53INP1 3′UTR, which contained the WT potential binding sites was synthesized and the MU fragments were generated by replacing G with T and A with C on the putative binding sites. (B) The target gene was verified by luciferase reporter assay. Co-transfection with miR-205 and TP53INP1-3′UTR significantly decreased the luciferase activity by 50.9% (P=0.006), 36.4% (P=0.03) and 43.9% (P=0.006) in CNE2, 6-10B and 9-4E cells, respectively. However, the activity of the reporter containing the mutated seed sequence was not obviously changed. Values are expressed as the mean ± standard deviation. *P<0.05 vs. NC. LUC, luciferase; WT, wild-type; Mut; mutant; UTR, untranslated region; miR, microRNA; NC, negative control; TP53INP1, tumor protein p53-inducible nuclear protein 1.

Mentions: Bioinformatics analysis with miRWalk (http://www.umm.uni-heidelberg.de/apps/zmf/mirwalk/) was used to identify potential targets of miR-205, and from these results, TP53INP1 was selected as a potential target for further study. To verify whether miR-205 directly targets TP53INP1, dual-luciferase reporter assays were performed. The sequences of the short fragments are shown in Fig. 6A. As shown in Fig. 6B, co-transfection of CNE2, 6-10B and 9-4E cells with TP53INP1-3′UTR and miR-205 mimic caused a significant decrease in the luciferase activity compared with that of the negative control. By contrast, the activity of the reporter containing the mutated seed sequence showed no obvious change in fluorescence. This result indicated that miR-205 exerts an inhibitory effect on TP53INP1 expression via binding to the 3′-UTR of TP53INP1, and that TP53INP1 is therefore a direct target of miR-205.


MicroRNA‑205 promotes the tumorigenesis of nasopharyngeal carcinoma through targeting tumor protein p53-inducible nuclear protein 1.

Nie G, Duan H, Li X, Yu Z, Luo L, Lu R, Ji Z, Zhang W - Mol Med Rep (2015)

TP53IPN1 is a target gene of miR-205. (A) Fragments of TP53INP1 3′UTR, which contained the WT potential binding sites was synthesized and the MU fragments were generated by replacing G with T and A with C on the putative binding sites. (B) The target gene was verified by luciferase reporter assay. Co-transfection with miR-205 and TP53INP1-3′UTR significantly decreased the luciferase activity by 50.9% (P=0.006), 36.4% (P=0.03) and 43.9% (P=0.006) in CNE2, 6-10B and 9-4E cells, respectively. However, the activity of the reporter containing the mutated seed sequence was not obviously changed. Values are expressed as the mean ± standard deviation. *P<0.05 vs. NC. LUC, luciferase; WT, wild-type; Mut; mutant; UTR, untranslated region; miR, microRNA; NC, negative control; TP53INP1, tumor protein p53-inducible nuclear protein 1.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4581759&req=5

f6-mmr-12-04-5715: TP53IPN1 is a target gene of miR-205. (A) Fragments of TP53INP1 3′UTR, which contained the WT potential binding sites was synthesized and the MU fragments were generated by replacing G with T and A with C on the putative binding sites. (B) The target gene was verified by luciferase reporter assay. Co-transfection with miR-205 and TP53INP1-3′UTR significantly decreased the luciferase activity by 50.9% (P=0.006), 36.4% (P=0.03) and 43.9% (P=0.006) in CNE2, 6-10B and 9-4E cells, respectively. However, the activity of the reporter containing the mutated seed sequence was not obviously changed. Values are expressed as the mean ± standard deviation. *P<0.05 vs. NC. LUC, luciferase; WT, wild-type; Mut; mutant; UTR, untranslated region; miR, microRNA; NC, negative control; TP53INP1, tumor protein p53-inducible nuclear protein 1.
Mentions: Bioinformatics analysis with miRWalk (http://www.umm.uni-heidelberg.de/apps/zmf/mirwalk/) was used to identify potential targets of miR-205, and from these results, TP53INP1 was selected as a potential target for further study. To verify whether miR-205 directly targets TP53INP1, dual-luciferase reporter assays were performed. The sequences of the short fragments are shown in Fig. 6A. As shown in Fig. 6B, co-transfection of CNE2, 6-10B and 9-4E cells with TP53INP1-3′UTR and miR-205 mimic caused a significant decrease in the luciferase activity compared with that of the negative control. By contrast, the activity of the reporter containing the mutated seed sequence showed no obvious change in fluorescence. This result indicated that miR-205 exerts an inhibitory effect on TP53INP1 expression via binding to the 3′-UTR of TP53INP1, and that TP53INP1 is therefore a direct target of miR-205.

Bottom Line: The present study aimed to explore the potential role of miR‑205 in NPC.The expression of miR‑205 was increased in NPC tissues compared with that in normal tissues.Overexpression of miR‑205 was found to promote the proliferation, migration and invasion of NPC‑derived cells, while apoptosis was suppressed.

View Article: PubMed Central - PubMed

Affiliation: Department of Otolaryngology, Peking University Shenzhen Hospital, Shenzhen, Guangdong 518036, P.R. China.

ABSTRACT
Nasopharyngeal carcinoma (NPC) is a common type of cancer in southern China, miRNAs have been shown to be involved in the tumorigenesis of multiple cancer types. The present study aimed to explore the potential role of miR‑205 in NPC. Reverse transcription quantitative polymerase chain reaction was used to determine the expression levels of miR‑205 in 20 fresh NPC specimens and 20 normal nasopharyngeal tissues. The function of miR‑205 in the proliferation, migration, invasion and apoptosis of NPC‑derived cells was detected by MTT assay, colony formation assay, wound healing assay, Transwell assay and flow cytometry. Furthermore, a target gene of miR‑205 was identified using the luciferase reporter assay. The expression of miR‑205 was increased in NPC tissues compared with that in normal tissues. Overexpression of miR‑205 was found to promote the proliferation, migration and invasion of NPC‑derived cells, while apoptosis was suppressed. Tumor protein p53-inducible nuclear protein 1 was identified as a target gene of miR‑205. Overall, the present study demonstrated that miR‑205 may function as an oncogene in NPC tumorigenesis.

Show MeSH
Related in: MedlinePlus