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Up-regulation of miR-9 target CBX7 to regulate invasion ability of bladder transitional cell carcinoma.

Xie D, Shang C, Zhang H, Guo Y, Tong X - Med. Sci. Monit. (2015)

Bottom Line: The expression of miR-9 increased significantly in human TCC specimens compared to that in NBTC specimens.The pcDNA-CBX7 transfection could up-regulate the CBX7 protein expression, and the invasion ability of T24 cells with CBX7 over-expression decreased significantly.Aberrantly expressed miR-9 contributes to T24 cells invasion, partly through directly down-regulating CBX7 protein expression in TCC.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, College of Basic Medicine, China Medical University, Shenyang, Liaoning, China (mainland).

ABSTRACT

Background: Bladder urothelial carcinoma is the most common genitourinary system cancer in China. The objective of this study was to investigate whether the miR-9 can regulate the invasion ability of human bladder transitional cell carcinoma cells by down-regulation of CBX7.

Material/methods: The expression of miR-9 was detected by quantitative real-time PCR in bladder transitional cell carcinomas (TCC) and normal bladder transitional cell (NBTC) samples. Bioinformatics software was used to predict some potential target genes of miR-9. T24 cells were transfected with pre-miR-9, and the CBX7 protein expression was detected by Western blot. Luciferase activities assay was selected to verify that CBX7 was a direct and specific gene of miR-9. T24 cells were transfected with pcDNA-CBX7, and the expression of CBX7 gene was detected. Then, the transwell assay was used to detect the invasion ability of T24 cells with CBX7 over-expression.

Results: The expression of miR-9 increased significantly in human TCC specimens compared to that in NBTC specimens. TargetScan and PicTar software programs predicted CBX7 gene was a target gene of miR-9. The pre-miR-9 could up-regulate the miR-9 expression and down-regulate CBX7 protein expression. The luciferase activities assay verified that CBX7 gene was a direct and specific target gene of miR-9. The pcDNA-CBX7 transfection could up-regulate the CBX7 protein expression, and the invasion ability of T24 cells with CBX7 over-expression decreased significantly.

Conclusions: Aberrantly expressed miR-9 contributes to T24 cells invasion, partly through directly down-regulating CBX7 protein expression in TCC. This miRNA signature offers a new potential therapeutic target for TCC.

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Related in: MedlinePlus

(A) 3′UTR of CBX7 is a target of miR-9 predicted by TargetScan and PicTar. (B) The luciferase reporter assay results, with each bar representing values from 3 independent experiments. The transfection efficiency was normalized by co-transfected Renilla luciferase and the light units were calculated by relative luciferase activity of firefly to Renilla. * P<0.01. (C) Representative image of the protein level of CBX7. GAPDH was used as a reference control. (D) Quantitative analysis of the relative protein levels of CBX7 normalized to those of GAPDH is shown. Data are mean ± SD of 3 independent experiments. * P<0.01.
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f2-medscimonit-21-225: (A) 3′UTR of CBX7 is a target of miR-9 predicted by TargetScan and PicTar. (B) The luciferase reporter assay results, with each bar representing values from 3 independent experiments. The transfection efficiency was normalized by co-transfected Renilla luciferase and the light units were calculated by relative luciferase activity of firefly to Renilla. * P<0.01. (C) Representative image of the protein level of CBX7. GAPDH was used as a reference control. (D) Quantitative analysis of the relative protein levels of CBX7 normalized to those of GAPDH is shown. Data are mean ± SD of 3 independent experiments. * P<0.01.

Mentions: TargetScan and PicTar predicted that the CBX7 gene was a target gene of miR-9. To determine the interaction between miR-9 and CBX7 gene, the luciferase reporter assay was used. The pre-miR-9 and reporter vectors were cotransfected into T24 cells. The relative luciferase activity in T24 cells decreased with WT vector by enhanced miR-9 level, and the above-mentioned inhibitive effect could be recovered with Mut vector. These results suggest that the CBX7 gene was a specific and direct target gene of miR-9 (Figure 2A, 2B).


Up-regulation of miR-9 target CBX7 to regulate invasion ability of bladder transitional cell carcinoma.

Xie D, Shang C, Zhang H, Guo Y, Tong X - Med. Sci. Monit. (2015)

(A) 3′UTR of CBX7 is a target of miR-9 predicted by TargetScan and PicTar. (B) The luciferase reporter assay results, with each bar representing values from 3 independent experiments. The transfection efficiency was normalized by co-transfected Renilla luciferase and the light units were calculated by relative luciferase activity of firefly to Renilla. * P<0.01. (C) Representative image of the protein level of CBX7. GAPDH was used as a reference control. (D) Quantitative analysis of the relative protein levels of CBX7 normalized to those of GAPDH is shown. Data are mean ± SD of 3 independent experiments. * P<0.01.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4307688&req=5

f2-medscimonit-21-225: (A) 3′UTR of CBX7 is a target of miR-9 predicted by TargetScan and PicTar. (B) The luciferase reporter assay results, with each bar representing values from 3 independent experiments. The transfection efficiency was normalized by co-transfected Renilla luciferase and the light units were calculated by relative luciferase activity of firefly to Renilla. * P<0.01. (C) Representative image of the protein level of CBX7. GAPDH was used as a reference control. (D) Quantitative analysis of the relative protein levels of CBX7 normalized to those of GAPDH is shown. Data are mean ± SD of 3 independent experiments. * P<0.01.
Mentions: TargetScan and PicTar predicted that the CBX7 gene was a target gene of miR-9. To determine the interaction between miR-9 and CBX7 gene, the luciferase reporter assay was used. The pre-miR-9 and reporter vectors were cotransfected into T24 cells. The relative luciferase activity in T24 cells decreased with WT vector by enhanced miR-9 level, and the above-mentioned inhibitive effect could be recovered with Mut vector. These results suggest that the CBX7 gene was a specific and direct target gene of miR-9 (Figure 2A, 2B).

Bottom Line: The expression of miR-9 increased significantly in human TCC specimens compared to that in NBTC specimens.The pcDNA-CBX7 transfection could up-regulate the CBX7 protein expression, and the invasion ability of T24 cells with CBX7 over-expression decreased significantly.Aberrantly expressed miR-9 contributes to T24 cells invasion, partly through directly down-regulating CBX7 protein expression in TCC.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, College of Basic Medicine, China Medical University, Shenyang, Liaoning, China (mainland).

ABSTRACT

Background: Bladder urothelial carcinoma is the most common genitourinary system cancer in China. The objective of this study was to investigate whether the miR-9 can regulate the invasion ability of human bladder transitional cell carcinoma cells by down-regulation of CBX7.

Material/methods: The expression of miR-9 was detected by quantitative real-time PCR in bladder transitional cell carcinomas (TCC) and normal bladder transitional cell (NBTC) samples. Bioinformatics software was used to predict some potential target genes of miR-9. T24 cells were transfected with pre-miR-9, and the CBX7 protein expression was detected by Western blot. Luciferase activities assay was selected to verify that CBX7 was a direct and specific gene of miR-9. T24 cells were transfected with pcDNA-CBX7, and the expression of CBX7 gene was detected. Then, the transwell assay was used to detect the invasion ability of T24 cells with CBX7 over-expression.

Results: The expression of miR-9 increased significantly in human TCC specimens compared to that in NBTC specimens. TargetScan and PicTar software programs predicted CBX7 gene was a target gene of miR-9. The pre-miR-9 could up-regulate the miR-9 expression and down-regulate CBX7 protein expression. The luciferase activities assay verified that CBX7 gene was a direct and specific target gene of miR-9. The pcDNA-CBX7 transfection could up-regulate the CBX7 protein expression, and the invasion ability of T24 cells with CBX7 over-expression decreased significantly.

Conclusions: Aberrantly expressed miR-9 contributes to T24 cells invasion, partly through directly down-regulating CBX7 protein expression in TCC. This miRNA signature offers a new potential therapeutic target for TCC.

Show MeSH
Related in: MedlinePlus