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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

Inhibition of miR-1246 reduced migration and invasion of SMMC7721 and BEL7402. (A, B) Transwell migration (n = 4) and invasion (n = 4) assays showed that SMMC7721 cells transfected with the miR-1246 inhibitor (800 nM) had lower invasive and migratory potentials than the control (inhibitor 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 transfected with the miR-1246 inhibitor (800 nM) had lower invasive and migratory potentials than the control (inhibitor 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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Fig1: Inhibition of miR-1246 reduced migration and invasion of SMMC7721 and BEL7402. (A, B) Transwell migration (n = 4) and invasion (n = 4) assays showed that SMMC7721 cells transfected with the miR-1246 inhibitor (800 nM) had lower invasive and migratory potentials than the control (inhibitor 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 transfected with the miR-1246 inhibitor (800 nM) had lower invasive and migratory potentials than the control (inhibitor 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: The higher expression of miR-1246 in more metastatic HCC line Hep12 promoted us to investigate its effect on the migration and invasion of HCC. BEL7402 and SMMC7721 cells were transfected with either miR-1246 mimic, mimic control, miR-1246 inhibitor or inhibitor control and then subjected to cell migration assay and cell invasion assay, respectively As expected, cell motility in both cell lines was significantly reduced after transfection of the miR-1246 inhibitor compared with inhibitor control (Figure 1). Similar results were obtained in Hep12 cells. Their motility was significantly reduced after transfection of the miR-1246 inhibitor compared with inhibitor control (Additional file 4: Figure S3). However, no difference was observed between miR-1246 mimic and mimic control in HCC line SMMC7721 and BEL7402 (Additional file 5: Figure S4). We speculate that this unexpected phenomenon may be caused by the relatively high expression of miR-1246 in SMMC7721 and BEL7402. So Hep11 with low expression of miR-1246 was chosen to evaluate the effects of miR-1246 mimic. Compared with control group, Hep11 transfected with miR-1246 mimic have significantly higher migration and invasion capacity (Figure 2). These results suggested that down-regulation of miR-1246 impaired migration and invasion of HCC while up-regulation of miR-1246 promoted migration and invasion.Figure 1


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)

Inhibition of miR-1246 reduced migration and invasion of SMMC7721 and BEL7402. (A, B) Transwell migration (n = 4) and invasion (n = 4) assays showed that SMMC7721 cells transfected with the miR-1246 inhibitor (800 nM) had lower invasive and migratory potentials than the control (inhibitor 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 transfected with the miR-1246 inhibitor (800 nM) had lower invasive and migratory potentials than the control (inhibitor 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.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4150976&req=5

Fig1: Inhibition of miR-1246 reduced migration and invasion of SMMC7721 and BEL7402. (A, B) Transwell migration (n = 4) and invasion (n = 4) assays showed that SMMC7721 cells transfected with the miR-1246 inhibitor (800 nM) had lower invasive and migratory potentials than the control (inhibitor 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 transfected with the miR-1246 inhibitor (800 nM) had lower invasive and migratory potentials than the control (inhibitor 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: The higher expression of miR-1246 in more metastatic HCC line Hep12 promoted us to investigate its effect on the migration and invasion of HCC. BEL7402 and SMMC7721 cells were transfected with either miR-1246 mimic, mimic control, miR-1246 inhibitor or inhibitor control and then subjected to cell migration assay and cell invasion assay, respectively As expected, cell motility in both cell lines was significantly reduced after transfection of the miR-1246 inhibitor compared with inhibitor control (Figure 1). Similar results were obtained in Hep12 cells. Their motility was significantly reduced after transfection of the miR-1246 inhibitor compared with inhibitor control (Additional file 4: Figure S3). However, no difference was observed between miR-1246 mimic and mimic control in HCC line SMMC7721 and BEL7402 (Additional file 5: Figure S4). We speculate that this unexpected phenomenon may be caused by the relatively high expression of miR-1246 in SMMC7721 and BEL7402. So Hep11 with low expression of miR-1246 was chosen to evaluate the effects of miR-1246 mimic. Compared with control group, Hep11 transfected with miR-1246 mimic have significantly higher migration and invasion capacity (Figure 2). These results suggested that down-regulation of miR-1246 impaired migration and invasion of HCC while up-regulation of miR-1246 promoted migration and invasion.Figure 1

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