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, a...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-nega- tive 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 LipofectamineTM2000. After 48 h, the levels of MTDH, E-cadherin and a-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 a-SMA was reduced and E-candherin were increased in MTDH-shRNA, with statisticat significance. Conclusion： Downregulation of MTDH increase E-candherin expression and reduced a-SMA expression, which inhibit EMT in MDA-MB-231 cells. This knockdown significantly suppresse migration and invasion in MDA-MB-231 cells.展开更多
基金Supported by grants from the Sub-topics of Major Discovery Platform in the 12th Five-Year Program of China (No. 2012ZX09303016-002)the Liaoning Province Science & Technology Department Funds (No. 201002060)
文摘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-nega- tive 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 LipofectamineTM2000. After 48 h, the levels of MTDH, E-cadherin and a-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 a-SMA was reduced and E-candherin were increased in MTDH-shRNA, with statisticat significance. Conclusion： Downregulation of MTDH increase E-candherin expression and reduced a-SMA expression, which inhibit EMT in MDA-MB-231 cells. This knockdown significantly suppresse migration and invasion in MDA-MB-231 cells.
