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MicroRNA-1 (miR-1) inhibits gastric cancer cell proliferation and migration by targeting MET.

Han C, Zhou Y, An Q, Li F, Li D, Zhang X, Yu Z, Zheng L, Duan Z, Kan Q - Tumour Biol. (2015)

Bottom Line: MicroRNAs (miRs) are short endogenous non-coding RNAs that act as posttranscriptional regulatory factors of gene expression.Cell protein expression of the miR-1 target gene MET was analyzed by Western blotting.MET siRNA also inhibited proliferation and migration in both cell lines.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical Pharmacology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, People's Republic of China.

ABSTRACT
MicroRNAs (miRs) are short endogenous non-coding RNAs that act as posttranscriptional regulatory factors of gene expression. Downregulation of miR-1 has been reported in gastric cancer; however, the mechanisms underlying its functions via target genes in gastric cancer remain largely unknown. The purpose of this study was to investigate the mechanism by which miR-1 inhibits gastric cancer cell proliferation and migration. The effects of miR-1 on gastric cancer cell proliferation and migration were determined by MTT and wound-healing assays. Cell protein expression of the miR-1 target gene MET was analyzed by Western blotting. Finally, MET expression was evaluated by immunohistochemistry in a stomach tumor tissue microarray (TMA). Ectopic expression of miR-1 inhibited proliferation and migration in both AGS and SGC-7901 gastric cancer cell lines. miR-1 directly targets the MET gene and downregulates its expression. MET siRNA also inhibited proliferation and migration in both cell lines. Immunohistochemistry revealed significantly higher MET expression levels in gastric cancer tissues compared with matched adjacent non-cancer tissues. These findings indicate that the miR-1/MET pathway is a potential therapeutic target due to its crucial role in gastric cancer cell proliferation and migration.

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Ectopic expression of miR-1 reduces cell motility in the wound scratch assay. a AGS cells were seeded in six-well plates at 2 × 105 cells per well. Wounds were generated using a micropipette tip upon cell adherence. Then, cells were transfected with miR-1 and non-specific miR mimics. The extent of wound healing was monitored by phase contrast microscopy, and photomicrographs were acquired at 48 and 72 h. b, c Quantification of cell migration using the monolayer wound-healing assay. The data were then analyzed using Prism 5.0 software and expressed as mean ± SEM
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Fig2: Ectopic expression of miR-1 reduces cell motility in the wound scratch assay. a AGS cells were seeded in six-well plates at 2 × 105 cells per well. Wounds were generated using a micropipette tip upon cell adherence. Then, cells were transfected with miR-1 and non-specific miR mimics. The extent of wound healing was monitored by phase contrast microscopy, and photomicrographs were acquired at 48 and 72 h. b, c Quantification of cell migration using the monolayer wound-healing assay. The data were then analyzed using Prism 5.0 software and expressed as mean ± SEM

Mentions: To understand the potential function of miR-1 in gastric cancer, MTT assay was carried out to assess the proliferation of gastric cancer cells after transfection with miR-1 mimics. As shown in Fig. 1a, b, the expression levels of miR-1 were upregulated in these two gastric cancer cell lines as confirmed by SYBR Green real-time RT-PCR. Then we performed MTT to determine the function of miR-1. The results showed that the two gastric cancer cell lines were significantly inhibited in miR-1 transfectants in comparison with cells transfected with the non-specific miR negative control (Fig. 1c, d). These results indicated that overexpression of miR-1 inhibited proliferation of gastric cancer cells in a dose-dependent manner in vitro (Fig. 1). Furthermore, wound-healing assay data showed significantly different widths for the residual gaps obtained in the miR-1 transfection and non-specific miR control groups, especially at 72 h (Fig. 2, P < 0.001). This experiment was repeated for three times.Fig. 1


MicroRNA-1 (miR-1) inhibits gastric cancer cell proliferation and migration by targeting MET.

Han C, Zhou Y, An Q, Li F, Li D, Zhang X, Yu Z, Zheng L, Duan Z, Kan Q - Tumour Biol. (2015)

Ectopic expression of miR-1 reduces cell motility in the wound scratch assay. a AGS cells were seeded in six-well plates at 2 × 105 cells per well. Wounds were generated using a micropipette tip upon cell adherence. Then, cells were transfected with miR-1 and non-specific miR mimics. The extent of wound healing was monitored by phase contrast microscopy, and photomicrographs were acquired at 48 and 72 h. b, c Quantification of cell migration using the monolayer wound-healing assay. The data were then analyzed using Prism 5.0 software and expressed as mean ± SEM
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4644207&req=5

Fig2: Ectopic expression of miR-1 reduces cell motility in the wound scratch assay. a AGS cells were seeded in six-well plates at 2 × 105 cells per well. Wounds were generated using a micropipette tip upon cell adherence. Then, cells were transfected with miR-1 and non-specific miR mimics. The extent of wound healing was monitored by phase contrast microscopy, and photomicrographs were acquired at 48 and 72 h. b, c Quantification of cell migration using the monolayer wound-healing assay. The data were then analyzed using Prism 5.0 software and expressed as mean ± SEM
Mentions: To understand the potential function of miR-1 in gastric cancer, MTT assay was carried out to assess the proliferation of gastric cancer cells after transfection with miR-1 mimics. As shown in Fig. 1a, b, the expression levels of miR-1 were upregulated in these two gastric cancer cell lines as confirmed by SYBR Green real-time RT-PCR. Then we performed MTT to determine the function of miR-1. The results showed that the two gastric cancer cell lines were significantly inhibited in miR-1 transfectants in comparison with cells transfected with the non-specific miR negative control (Fig. 1c, d). These results indicated that overexpression of miR-1 inhibited proliferation of gastric cancer cells in a dose-dependent manner in vitro (Fig. 1). Furthermore, wound-healing assay data showed significantly different widths for the residual gaps obtained in the miR-1 transfection and non-specific miR control groups, especially at 72 h (Fig. 2, P < 0.001). This experiment was repeated for three times.Fig. 1

Bottom Line: MicroRNAs (miRs) are short endogenous non-coding RNAs that act as posttranscriptional regulatory factors of gene expression.Cell protein expression of the miR-1 target gene MET was analyzed by Western blotting.MET siRNA also inhibited proliferation and migration in both cell lines.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical Pharmacology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, People's Republic of China.

ABSTRACT
MicroRNAs (miRs) are short endogenous non-coding RNAs that act as posttranscriptional regulatory factors of gene expression. Downregulation of miR-1 has been reported in gastric cancer; however, the mechanisms underlying its functions via target genes in gastric cancer remain largely unknown. The purpose of this study was to investigate the mechanism by which miR-1 inhibits gastric cancer cell proliferation and migration. The effects of miR-1 on gastric cancer cell proliferation and migration were determined by MTT and wound-healing assays. Cell protein expression of the miR-1 target gene MET was analyzed by Western blotting. Finally, MET expression was evaluated by immunohistochemistry in a stomach tumor tissue microarray (TMA). Ectopic expression of miR-1 inhibited proliferation and migration in both AGS and SGC-7901 gastric cancer cell lines. miR-1 directly targets the MET gene and downregulates its expression. MET siRNA also inhibited proliferation and migration in both cell lines. Immunohistochemistry revealed significantly higher MET expression levels in gastric cancer tissues compared with matched adjacent non-cancer tissues. These findings indicate that the miR-1/MET pathway is a potential therapeutic target due to its crucial role in gastric cancer cell proliferation and migration.

Show MeSH
Related in: MedlinePlus