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MicroRNA-1246 enhances migration and invasion through CADM1 in hepatocellular carcinoma.

Sun Z, Meng C, Wang S, Zhou N, Guan M, Bai C, Lu S, Han Q, Zhao RC - BMC Cancer (2014)

Bottom Line: The aberrant expression of microRNAs has been demonstrated to play a crucial role in the initiation and progression of hepatocarcinoma. miR-1246 expression in High invasive ability cell line than significantly higher than that in low invasive ability cell line.Inhibition of miR-1246 effectively reduced migration and invasion of hepatocellular carcinoma cell lines.Our study showed that miR-1246, by down-regulation CADM1, enhances migration and invasion in HCC cells.

View Article: PubMed Central - PubMed

Affiliation: Institute of Basic Medical Sciences Chinese Academy of Medical Sciences, School of Basic Medicine Peking Union Medical College, Beijing, People's Republic of China. hanqinhanqin@126.com.

ABSTRACT

Background: The aberrant expression of microRNAs has been demonstrated to play a crucial role in the initiation and progression of hepatocarcinoma. miR-1246 expression in High invasive ability cell line than significantly higher than that in low invasive ability cell line.

Methods: Transwell chambers (8-uM pore size; Costar) were used in the in vitro migration and invison anssay. Dual luciferase reporter gene construct and Dual luciferase reporter assay to identify the target of miR-1246. CADM1 expression was evaluated by immunohistochemistric staining. The clinical manifestations, treatments and survival were collected for statistical analysis.

Results: Inhibition of miR-1246 effectively reduced migration and invasion of hepatocellular carcinoma cell lines. Bioinformatics and luciferase reporter assay revealed that miR-1246 specifically targeted the 3'-UTR of Cell adhesion molecule 1 and regulated its expression. Down-regulation of CADM1 enhanced migration and invasion of HCC cell lines. Furthermore, in tumor tissues obtained from liver cancer patients, the expression of miR-1246 was negatively correlated with CADM1 and the high expression of miR-1246 combined with low expression of CADM1 might serve as a risk factor for stage1 liver cancer patients.

Conclusions: Our study showed that miR-1246, by down-regulation CADM1, enhances migration and invasion in HCC cells.

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Related in: MedlinePlus

CADM1 regulated HCC cell migration and invasion. (A, B) Transwell migration (n = 4) and invasion (n = 4) assays showed that SMMC7721 cells that were transfected with the CADM1 siRNA (200 nM) had greater invasive and migratory potentials than the control (siRNA control). (A) is a microscopic image of Crystal violet staining; (B) shows the statistical results. (C, D) Transwell migration (n = 4) and invasion (n = 4) assays showed that BEL7402 cells that were transfected with the CADM1 siRNA (200 nM) had greater invasive and migratory potentials than the control (siRNA control). (C) is a microscopic image of Crystal violet staining; (D) shows the statistical results. Data represent the mean ± SD of four independent experiments. *P < 0.01.
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Fig4: CADM1 regulated HCC cell migration and invasion. (A, B) Transwell migration (n = 4) and invasion (n = 4) assays showed that SMMC7721 cells that were transfected with the CADM1 siRNA (200 nM) had greater invasive and migratory potentials than the control (siRNA control). (A) is a microscopic image of Crystal violet staining; (B) shows the statistical results. (C, D) Transwell migration (n = 4) and invasion (n = 4) assays showed that BEL7402 cells that were transfected with the CADM1 siRNA (200 nM) had greater invasive and migratory potentials than the control (siRNA control). (C) is a microscopic image of Crystal violet staining; (D) shows the statistical results. Data represent the mean ± SD of four independent experiments. *P < 0.01.

Mentions: We next evaluated whether downregulation of CADM1 levels might have a reverse effect on cell motility as observed following miR-1246 inhibition. Small interfering RNA (siRNA) designed to target at CADM1 was employed. Transfection of CADM1 siRNA caused a more than 50% reduction in CADM1 protein level compared to the Si-control (Additional file 9: Figure S8). We then compared expression of CADM1 in different HCC cell lines and found SMMC7721 and BEL7402 cells have high CADM1 expression levels (Additional file 10: Figure S9). These two cell lines were transfected with CADM1 siRNA and subjected to cell migration assay and cell invasion assay. As shown in Figure 4, cell motility was significantly enhanced after CADM1 downregulation compared with the control. Similar results were obtained in Hep11 cells (Additional file 11: Figure S10).Figure 4


MicroRNA-1246 enhances migration and invasion through CADM1 in hepatocellular carcinoma.

Sun Z, Meng C, Wang S, Zhou N, Guan M, Bai C, Lu S, Han Q, Zhao RC - BMC Cancer (2014)

CADM1 regulated HCC cell migration and invasion. (A, B) Transwell migration (n = 4) and invasion (n = 4) assays showed that SMMC7721 cells that were transfected with the CADM1 siRNA (200 nM) had greater invasive and migratory potentials than the control (siRNA control). (A) is a microscopic image of Crystal violet staining; (B) shows the statistical results. (C, D) Transwell migration (n = 4) and invasion (n = 4) assays showed that BEL7402 cells that were transfected with the CADM1 siRNA (200 nM) had greater invasive and migratory potentials than the control (siRNA control). (C) is a microscopic image of Crystal violet staining; (D) shows the statistical results. Data represent the mean ± SD of four independent experiments. *P < 0.01.
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Related In: Results  -  Collection

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Fig4: CADM1 regulated HCC cell migration and invasion. (A, B) Transwell migration (n = 4) and invasion (n = 4) assays showed that SMMC7721 cells that were transfected with the CADM1 siRNA (200 nM) had greater invasive and migratory potentials than the control (siRNA control). (A) is a microscopic image of Crystal violet staining; (B) shows the statistical results. (C, D) Transwell migration (n = 4) and invasion (n = 4) assays showed that BEL7402 cells that were transfected with the CADM1 siRNA (200 nM) had greater invasive and migratory potentials than the control (siRNA control). (C) is a microscopic image of Crystal violet staining; (D) shows the statistical results. Data represent the mean ± SD of four independent experiments. *P < 0.01.
Mentions: We next evaluated whether downregulation of CADM1 levels might have a reverse effect on cell motility as observed following miR-1246 inhibition. Small interfering RNA (siRNA) designed to target at CADM1 was employed. Transfection of CADM1 siRNA caused a more than 50% reduction in CADM1 protein level compared to the Si-control (Additional file 9: Figure S8). We then compared expression of CADM1 in different HCC cell lines and found SMMC7721 and BEL7402 cells have high CADM1 expression levels (Additional file 10: Figure S9). These two cell lines were transfected with CADM1 siRNA and subjected to cell migration assay and cell invasion assay. As shown in Figure 4, cell motility was significantly enhanced after CADM1 downregulation compared with the control. Similar results were obtained in Hep11 cells (Additional file 11: Figure S10).Figure 4

Bottom Line: The aberrant expression of microRNAs has been demonstrated to play a crucial role in the initiation and progression of hepatocarcinoma. miR-1246 expression in High invasive ability cell line than significantly higher than that in low invasive ability cell line.Inhibition of miR-1246 effectively reduced migration and invasion of hepatocellular carcinoma cell lines.Our study showed that miR-1246, by down-regulation CADM1, enhances migration and invasion in HCC cells.

View Article: PubMed Central - PubMed

Affiliation: Institute of Basic Medical Sciences Chinese Academy of Medical Sciences, School of Basic Medicine Peking Union Medical College, Beijing, People's Republic of China. hanqinhanqin@126.com.

ABSTRACT

Background: The aberrant expression of microRNAs has been demonstrated to play a crucial role in the initiation and progression of hepatocarcinoma. miR-1246 expression in High invasive ability cell line than significantly higher than that in low invasive ability cell line.

Methods: Transwell chambers (8-uM pore size; Costar) were used in the in vitro migration and invison anssay. Dual luciferase reporter gene construct and Dual luciferase reporter assay to identify the target of miR-1246. CADM1 expression was evaluated by immunohistochemistric staining. The clinical manifestations, treatments and survival were collected for statistical analysis.

Results: Inhibition of miR-1246 effectively reduced migration and invasion of hepatocellular carcinoma cell lines. Bioinformatics and luciferase reporter assay revealed that miR-1246 specifically targeted the 3'-UTR of Cell adhesion molecule 1 and regulated its expression. Down-regulation of CADM1 enhanced migration and invasion of HCC cell lines. Furthermore, in tumor tissues obtained from liver cancer patients, the expression of miR-1246 was negatively correlated with CADM1 and the high expression of miR-1246 combined with low expression of CADM1 might serve as a risk factor for stage1 liver cancer patients.

Conclusions: Our study showed that miR-1246, by down-regulation CADM1, enhances migration and invasion in HCC cells.

Show MeSH
Related in: MedlinePlus