ABSTRACT
Objective To investigate the molecular mechanism of miR-139-5p targeting transforming growth factor-β1 (TGF-β1) in the regulation of epithelial mesenchymal transition (EMT),thus inhibiting invasion and metastasis of hepatoma cells.Methods Bioinformatics methods were used to determine whether miR-139-5p was the best binding miRNA of TGF-β1.Correlation between the TGF-β1 expression as detected by immunohistochemistry and Western blot,and the miR-139-5p level by qRT-PCR in 56 hepatoma tissues and 20 normal tissues,respectively,was analyzed.The relationship between the miR-139-5p level as detected by qRT-PCR,and TGF-β1,E-cadherin and Vimentin by Western blot in the high and low metastatic hepatoma cell lines were investigated.In recombinant cell lines,whether miR-139-5p could bind to the 3'UTR site of TGF-β1 was evaluated,and the effect on invasive ability after modulating miR-139-5p level was also tested by the transwell method.Results A total of 20 miRNAs were found to be able to bind with TGF-β1 by bioinformatics methods and among these mRNAs,miR-139-5p was the best target miRNA with the highest specificity and strongest stability to bind TGF-β1.The positive expression rates of TGF-β1 in hepatoma tissues and adjacent normal liver tissues were 80.4％ (45/56) and 15.0％ (3/20),respectively,(P ＜0.05).There were significant differences on the expressions of TGF-β1,E-cadherin and Vimentin among the different metastatic cell lines (all P ＜ 0.05).After miR-139-5p was transfected into hepatoma cells,miR-139-5p could bind to the 3'UTR site of TGF-β1,resulting in downregulating TGF-β1 expression.When compared to the other three groups,the cell line with a high expression of miR-139-5p had a significantly lower count of invasive cells (53 ± 4/high magnification field) (P ＜ 0.05).Conclusion miRNA139-5p could specifically bind to the 3'UTR site of TGF-β1 and regulate the EMT signaling pathway,thus suppressing invasion and metastasis of hepatoma cells.
ABSTRACT
Objective To investigate the clinical features and management of bile duct injury caused by laparoscopic cholecysteetomy by using harmonic scalpel (UHS-LC), and its prevention. Methods The clinical data of 1863 UHS-LC cases from April 2003 to February 2008 were retrospectively analyzed. There were 11 patients suffering from UHS-LC related iatrogenic bile duct injury including intraoperatively immediate recognized injuries in 9 cases, and postoperatively found injuries in 2 cases. For those patients in which bile duct injury was found during the UHS-LC procedure, the patient was converted to open surgery, the injury was repaired accordingly by end-to-end bile duet anastomosis or Roux-en-Y procedure. For the injuries found postoperatively (all two cases were of CBD perforation), CBD was sutured by second stage. Results All the 11 patients recovered well and no biliary stricture occurred during the follow up of 1-5 years. Conclusions While UHS-LC is suitable for most cases of choleeystectomy, it causes significant porcentage of bile duct injury (0. 6% ) in hands of unskillful surgeons. It is important to be on alert against iatrugenic bile duct injuries.