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Objective: Metadherin (MTDH) could regulate epithelial-mesenchymal transition (EMT), and is involved in tumor metastasis.This study was designed to observe the effect of MTDH-short hairpin RNA (shRNA) on EMT, and the role of MTDH in breast tumor metastasis. Methods: RNA interference plasmid that can express shRNA targeting MTDH or shRNA-negative plasmid that does not match any known human coding mRNA was designed, constructed and named MTDH-shRNA and MTDH-shRNA-neg, which were transiently transfected into MDA-MB-231 cells using Lipofectamine TM2000. After 48 h, the levels of MTDH, E-cadherin and α-SMA expression were determined by reverse transcription-polymerase chain reactin (RT-PCR), Western blot. The invasion and immigration potential was examined by Transwell chamber invasion or migration assay. Results: Compared with MDA-MB-231, the MTDH mRNA level was down-regulated by 41.2%, the MTDH protein level was down-regulated by 40.3%. The invasion and immigration potential of MDA-MB-231 cells was decreased after transfection of MTDH-shRNA. Compared with MDA-MB-231 or MTDH-shRNA-neg, the mRNA and protein level of α-SMA was reduced and E-candherin were increased in MTDH-shRNA, with statistical significance. Conclusion: Downregulation of MTDH increase E-candherin expression and reduced α-SMA expression, which inhibit EMT in MDA-MB-231 cells. This knockdown significantly suppresse migration and invasion in MDA-MB-231 cells.
Objective: Metadherin (MTDH) could regulate epithelial-mesenchymal transition (EMT), and is involved in tumor metastasis. This study was designed to observe the effect of MTDH-short hairpin RNA (shRNA) on EMT, and the role of MTDH in breast tumor metastasis. Methods: RNA interference plasmid that can express shRNA targeting MTDH or shRNA-negative plasmid that does not match any known human coding mRNA was designed, constructed and named MTDH-shRNA and MTDH-shRNA-neg, which were transiently transfected into MDA -MB-231 cells using Lipofectamine ™ 2000. After 48 h, the levels of MTDH, E-cadherin and α-SMA expression were determined by reverse transcription-polymerase chain reactin (RT-PCR), Western blot. The invasion and immigration potential was examined by Transwell chamber invasion or migration assay. Results: Compared with MDA-MB-231, the MTDH mRNA level was down-regulated by 41.2%, the MTDH protein level was down-regulated by 40.3%. The invasion and immigration potential of MDA -MB-23 1 cells was decreased after transfection of MTDH-shRNA. Compared with MDA-MB-231 or MTDH-shRNA-neg, the mRNA and protein level of α-SMA was reduced and E-candherin were increased in MTDH-shRNA, with the statistical significance . Conclusion: Downregulation of MTDH increase in E-candherin expression and reduced α-SMA expression, which inhibit EMT in MDA-MB-231 cells. This knockdown significantly suppresses migration and invasion in MDA-MB-231 cells.