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. 2018 Oct 4;18(1):952.
doi: 10.1186/s12885-018-4863-y.

Uc.416 + A promotes epithelial-to-mesenchymal transition through miR-153 in renal cell carcinoma

Yohei Sekino  1   2 , Naoya Sakamoto  1 , Keisuke Goto  3 , Ririno Honma  1 , Yoshinori Shigematsu  1   2 , Thang Pham Quoc  1 , Kazuhiro Sentani  1 , Naohide Oue  1 , Jun Teishima  2 , Fumi Kawakami  4 , Jose A Karam  5 , Kanishka Sircar  4   6 , Akio Matsubara  2 , Wataru Yasui  7
Affiliations

Uc.416 + A promotes epithelial-to-mesenchymal transition through miR-153 in renal cell carcinoma

Yohei Sekino et al. BMC Cancer. .

Abstract

Background: The transcribed ultraconserved regions (T-UCRs) are a novel class of non-coding RNAs that are absolutely conserved across species and are involved in carcinogenesis in some cancers. However, the expression and biological role of T-UCRs in renal cell carcinoma (RCC) remain poorly understood. This study aimed to examine the expression and functional role of Uc.416 + A and analyze the association between Uc.416 + A and epithelial-to-mesenchymal transition in RCC.

Methods: Expression of Uc.416 + A in 35 RCC tissues, corresponding normal kidney tissues and 13 types of normal tissue samples was determined by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). We performed a cell growth and migration assay in RCC cell line 786-O transfected with negative control and siRNA for Uc.416 + A. We evaluated the relation between Uc.416 + A and miR-153, which has a complimentary site of Uc.416 + A.

Results: qRT-PCR analysis revealed that the expression of Uc.416 + A was higher in RCC tissues than that in corresponding normal kidney tissues. Inhibition of Uc.416 + A reduced cell growth and cell migration activity. There was an inverse correlation between Uc.416 + A and miR-153. Western blot analysis showed Uc.416 + A modulated E-cadherin, vimentin and snail. The expression of Uc.416 + A was positively associated with the expression of SNAI1, VIM and inversely associated with the expression of CDH1.

Conclusions: The expression of Uc.416 + A was upregulated in RCC and especially in RCC tissues with sarcomatoid change. Uc.416 + A promoted epithelial-to-mesenchymal transition through miR-153. These results suggest that Uc.416 + A may be a promising therapeutic target.

Keywords: Epithelial-to-mesenchymal transition; Renal cell carcinoma; Sarcomatoid change; Uc.416 + A; miR-153.

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Conflict of interest statement

Ethics approval and consent to participate

35 RCC tissue samples were collected from patients at Hiroshima University Hospital or an affiliated hospital under an institutional review board-approved protocol (IRB# E912). The written comprehensive informed consent was obtained from all of the patients. This study was conducted in accordance with the Ethical Guidance for Human Genome/Gene Research of the Japanese Government. We also obtained 15 frozen sarcomatoid RCC tissue samples and adjacent normal kidney samples from the tissue bank of The University of Texas MD Anderson Cancer Center (Houston, TX) under approval of the written informed consent and using an institutional review board-approved protocol (IRB# LAB 08–670).

Consent for publication

Not applicable.

Competing interests

The author declare that they have no competing interests.

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Figures

Fig. 1
Fig. 1
Overexpression of Uc.416 + A in renal cell carcinoma tissues (RCC). a The results of qRT-PCR analysis for the expression of Uc.416 + A in 35 RCC tissues and corresponding normal kidney tissues (N). Statistical differences were evaluated with the paired T test. b Scatter plot diagrams showing the association between the expression of Uc.416 + A and metastatic status. Statistical differences were evaluated with the Mann-Whitney U-test. NM: non-metastatic RCC tissues. M: metastatic RCC tissues. c Results of qRT-PCR analysis for the expression of Uc.416 + A in 35 RCC tissues and 13 types of normal samples
Fig. 2
Fig. 2
Uc.416 + A is involved in cell proliferation and migration in 786-O cells. a The results of qRT-PCR analysis for the expression of Uc.416 + A in 786-O and Caki-1 cells. The results are expressed as the mean ± S.D. of triplicate measurements. b The results of qRT-PCR analysis for the expression of Uc.416 + A in 786-O cells transfected with negative control or two different siRNAs. The results are expressed as the mean ± S.D. of triplicate measurements. ***P < 0.001. c Cell proliferation assay in 786-O cells transfected with negative control or two different siRNAs. Cell growth was assessed by MTT assays at 1, 2, and 4 days after seeding on 96-well plates. Bars and error bars indicate the mean and S.D., respectively, of 3 independent experiments. **P < 0.01. d Wound healing assay in 786-O cells transfected with negative control or two different siRNAs. Wound closures were evaluated by wound contraction percentage and closure time at 0, 12, and 24 h after scratching. The results are expressed as the mean and S.D. of triplicate measurements. **P < 0.01
Fig. 3
Fig. 3
Interaction between Uc.416 + A and miR-153. a The results of qRT-PCR analysis for the expression of miR-153 in 32 renal cell carcinoma (RCC) tissues and corresponding normal kidney tissues (N). Statistical differences were evaluated with the paired T test. b The correlation between Uc.416 + A and miR-153 in RCC tissues. Spearman correlation coefficient and P-values are indicated. c qRT-PCR analysis for the expression of miR-153 in 786-O cells transfected with negative control or two different siRNAs. d qRT-PCR analysis for the expression of miR-153 in 786-O cells transfected with negative control or miR-153 mimics. The results are expressed as the mean and S.D. of triplicate measurements. ***P < 0.001. e qRT-PCR analysis for the expression of Uc.416 + A in 786-O cells transfected with negative control or miR-153 mimics. The results are expressed as the mean and S.D. of triplicate measurements. ***P < 0.001. f The luciferase activity of 786-O cells co-transfected with a control pGL3-Promoter vecor containing a wild type Uc.416 + A sequence or a pGL3-Promoter vector containing a mutated Uc.416 + A sequence. The results are expressed as the mean and S.D. of triplicate measurements. ***P < 0.001. N.S., not significant; WT: wild type; Mut: sequence with a mutated miR-153 binding site
Fig. 4
Fig. 4
Uc.416 + A modulates epithelial-to-mesenchymal transition. a Western blot analysis of E-cadherin, snail and vimentin in 786-O cells transfected with negative control or two different siRNAs for Uc.416 + A. β-actin was used as a loading control. b The relative expression level of Uc.416 + A in 35 RCC samples lacking sarcomatoid change and in 15 RCC samples with sarcomatoid change. c The correlation between the expression of Uc.416 + A and the ratio of sarcomatoid components in RCC samples with sarcomatoid change. Spearman correlation coefficient and P-values are indicated. d The correlation between the expression of Uc.416 + A and the expression of SNAI1 in RCC samples with sarcomatoid change. Spearman correlation coefficient and P-values are indicated. e The correlation between the expression of Uc.416 + A and the expression of VIM in RCC samples with sarcomatoid change. Spearman correlation coefficient and P-values are indicated. f The correlation between the expression of Uc.416 + A and the expression of CDH1 in RCC samples with sarcomatoid change. Spearman correlation coefficient and P-values are indicated
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