Abstract:
To enhance the cancer cell transfection efficiency by the optimized calcium phosphate method, the ANXA2 specific inhibition recombinant plasmid (pU6H1-GFP-siANXA2) was introduced into the human hepatocarcinoma cell, SMMC-7721. The following conditions were optimized: Cultured cell cover ratio before transfection, plasmid dose, DNA precipitation formation time, incubation time, cell growth status, and the absence/attendance of serum and antibiotics. The results indicated the optimal transfection conditions as the follows: 50%～60% cultured cell cover ratio before transfection, 8 μg plasmid per 30 mm dish, 20～30 min complex formation time, 6 h incubation time. The absence/attendance of antibiotics showed no significant influence. Serum was indispensable for this method. ANXA2 mRNA and protein level detection results indicated that the target gene expression was significantly inhibited at 48 h post transfection (P
