[Construction of human Bcl-6 3'UTR reporter vector and expression vector and their functional assessment].

OBJECTIVE: To observe the direct regulation of miR-127 on Bcl-6 and the effect of Bcl-6 in rescuing miR-127-induced cell cycle and cell growth inhibition. METHODS: The 3'UTR and coding region of human bcl-6 gene were amplified by PCR and cloned into pcDNA3.0-Luc and pcDNA3.0-Flag vectors, respectively. Mutations were introduced into the seed sequences of the predicted miR-127 target sites within the Bcl-6 3'UTR using recombinant PCR. Luciferase assay was used to verify the direct targeted regulation of miR-127 on Bcl-6. In HepG2 cell models with overexpression or knockdown of miR-12, the changes of cell cycle and cell growth were investigated after transfection with the constructed vectors. RESULTS: The recombinant plasmids were successfully obtained as confirmed by double digestion and sequence identification. Luciferase assay showed that in 293T and HepG2 cells, miR-127 inhibited the activation of wild-type Bcl-6 3'UTR reporter vector but not mutated Bcl-6 3'UTR vector. Overexpression of miR-127 induced cell cycle arrest at G(2)/M phase and suppressed the growth of HepG2 cells, and these effects were reversed by Bcl-6 overexpression. CONCLUSION: We successfully cloned wild-type and mutated 3'UTR reporter vectors and expression vector of bcl-6 gene and confirmed their biological functions.

Sorafenib combined with TACE in advanced primary hepatocellular carcinoma.

BACKGROUND/AIMS: To evaluate the effectiveness and safety of sorafenib combined with transarterial chemoembolization (TACE) in patients with advanced primary hepatocellular carcinoma. METHODOLOGY: A retrospective analysis of 65 patients with advanced primary hepatocellular carcinoma who had been administered sorafenib for more than one month and treated by TACE was performed. The tumor response was evaluated according to the response evaluation criteria in solid tumors and any side effects were recorded. RESULTS: Twelve patients entered a partial remission, 42 entered a stable condition, and the disease progressed in 11 patients. The disease control and objective regression rates were 83.1% and 18.5%, respectively. The one-year survival rate was 63.1%. Among the cases, the median overall survival time was 17 months, and the median time to progression was 9 months. The overall side effects rate was 78.5% and most were relieved after treatment. CONCLUSIONS: Sorafenib combined with TACE is a safe and effective treatment of advanced primary hepatocellular carcinoma.