摘要
OBJECTIVE To study the effect of the Twist gene on the migration of human hepatocellular carcinoma cells and the possible mechanisms involved. METHODS RT-PCR was used to detect expression of the Twist gene in primary (Hep11) and recurrent (Hep12) cell lines from the same HCC patient. Hep11 cells were stably transfected with Twist-cDNA, and Hep12 cells were transiently transfected with Twist RNAi plasmid. Cell migration assays were performed on Twist up-regulated Hep11 cells and Twist RNAi Hep12 cells. RT- PCR and Western blot were used to test the expression of EMT markers. RESULTS Twist was expressed higher level and had increased migration capability in recurrent Hep12 cells than those in primary Hep11 cells. Cell models (Twist-Hep11) in which Twist protein was steadily and highly expressed were obtained. Compared with pcDNA3-Hep11 cells, migration of Twist-Hep11 cells was clearly increased. However, migration of Twist RNAi (Si-Twist-Hep12) Hep12 cells were reduced. Overexpression of Twist in Hep11 cells promoted expression of N-cad and vimentin. CONCLUSION These results indicate that Twist promotes the migration of hepatocellular carcinoma cells in vitro and may play an important role in the upregulation of mesenchymal markers.
OBJECTIVE To study the effect of the Twist gene on themigration of human hepatocellular carcinoma cells and thepossible mechanisms involved.METHODS RT-PCR was used to detect expression of the Twistgene in primary (Hepl1) and recurrent (Hep12) cell lines fromthe same HCC patient.Hepll cells were stably transfected withTwist-cDNA,and Hep12 cells were transiently transfected withTwist RNAi plasmid.Cell migration assays were performed onTwist up-regulated Hepll cells and Twist RNAi Hep12 cells.RTPCRand Western blot were used to test the expression of EMTmarkers.RESULTS Twist was expressed higher level and had increasedmigration capability in recurrent Hep12 cells than those in primaryHepll cells.Cell models (Twist-Hep1l) in which Twist proteinwas steadily and highly expressed were obtained.Compared withpcDNA3-Hepll cells,migration of Twist-Hepll cells was clearlyincreased.However,migration of Twist RNAi (Si-Twist-Hepl2)Hep12 cells were reduced.Overexpression of Twist in Hep11 cellspromoted expression of N-cad and vimentin.CONCLUSION These results indicate that Twist promotes themigration of hepatocellular carcinoma cells in vitro and may playan important role in the upregulation of mesenchymal markers.
基金
supported by grants from the National Natural Science Foundation of China (No.30671060)
Program for New Century Excellent Talent in Universities,China (No.NCET-07-0031)