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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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MET siRNA inhibits AGS and SGC-7901 cell growth. a, c AGS cells were transfected with si-MET-1/2/3 mimics and non-specific siRNA mimics at 5–80 nM, respectively, and incubated for up to 72 h in a medium containing 10 % FBS. b SGC-7901 cells were transfected with si-MET-1/2/3 mimics and non-specific miR mimics at 5–80 nM and incubated for 72 h. Cell growth was measured as described above
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Fig5: MET siRNA inhibits AGS and SGC-7901 cell growth. a, c AGS cells were transfected with si-MET-1/2/3 mimics and non-specific siRNA mimics at 5–80 nM, respectively, and incubated for up to 72 h in a medium containing 10 % FBS. b SGC-7901 cells were transfected with si-MET-1/2/3 mimics and non-specific miR mimics at 5–80 nM and incubated for 72 h. Cell growth was measured as described above

Mentions: MET is known to be associated with carcinoma in various cancer types. The expression level of the MET protein was significantly decreased in both gastric cancer cell lines (AGS and SGC-7901) after transfection with miR-1 mimics (Fig. 3). In addition, the expression levels of survivin were also inhibited in both two cell lines (Fig. 3c, f). Conversely, MET expression was markedly reduced in MET siRNA (si-MET) transfectants. Specifically, inhibition of MET expression in AGS cells was more robust compared with that obtained for the SGC-7901 cell line (Fig. 4). AS expected, there are no significant changes on the expression levels of survivin in MET siRNA transfected cells. Loss-of-function assays using siRNA analysis were performed to examine the effect of MET on gastric cancer cell growth. The MTT assay revealed significant cell growth inhibition in si-MET-1/2/3 transfected cells after a 72-h transfection (Fig. 5).Fig. 4


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)

MET siRNA inhibits AGS and SGC-7901 cell growth. a, c AGS cells were transfected with si-MET-1/2/3 mimics and non-specific siRNA mimics at 5–80 nM, respectively, and incubated for up to 72 h in a medium containing 10 % FBS. b SGC-7901 cells were transfected with si-MET-1/2/3 mimics and non-specific miR mimics at 5–80 nM and incubated for 72 h. Cell growth was measured as described above
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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Fig5: MET siRNA inhibits AGS and SGC-7901 cell growth. a, c AGS cells were transfected with si-MET-1/2/3 mimics and non-specific siRNA mimics at 5–80 nM, respectively, and incubated for up to 72 h in a medium containing 10 % FBS. b SGC-7901 cells were transfected with si-MET-1/2/3 mimics and non-specific miR mimics at 5–80 nM and incubated for 72 h. Cell growth was measured as described above
Mentions: MET is known to be associated with carcinoma in various cancer types. The expression level of the MET protein was significantly decreased in both gastric cancer cell lines (AGS and SGC-7901) after transfection with miR-1 mimics (Fig. 3). In addition, the expression levels of survivin were also inhibited in both two cell lines (Fig. 3c, f). Conversely, MET expression was markedly reduced in MET siRNA (si-MET) transfectants. Specifically, inhibition of MET expression in AGS cells was more robust compared with that obtained for the SGC-7901 cell line (Fig. 4). AS expected, there are no significant changes on the expression levels of survivin in MET siRNA transfected cells. Loss-of-function assays using siRNA analysis were performed to examine the effect of MET on gastric cancer cell growth. The MTT assay revealed significant cell growth inhibition in si-MET-1/2/3 transfected cells after a 72-h transfection (Fig. 5).Fig. 4

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