Transfection of mIL-12 gene mediated by adenoviral vector to H22 cells and its effect on the growth of H22 cells

Aim: To study the transfection of AdmIL-12 and Ad-EGFP to H22 cells, which effect the growth change on H22 cells. Methods: The cDNA of p35 and p40 of mIL-12 gene were inserted into the region of E1 and E3 of Ad5 respectively. AdmIL-12 was valuated and viral tite was detected after AdmIL-12 was constructed. The recombinant adenovirus vector with enhanced green fluorescence protein gene (Ad-EGFP) was constructed by the same way. Ad-EGFP transfected H22 cells by different multiplicity of infection (MOI) and time of infection (TOI), then the transfection efficiency of Ad-EGFP was observed. H22 cells were transfected by AdmIL-12 and Ad-EGFP respectively, then the content of mIL-12 in the supernatant was detected by ELISA. The effects of AdmIL-12 and Ad-EGFP on H22 cells growth was detected by improved MTT. Results: AdmIL-12 and Ad-EGFP were obtained, and their plaque-forming units were 1×10~8/mL and 1×10~9/mL. The transfection efficiency of Ad-EGFP were 87.67%、96.38%、96.43% and 96.32% respectively when TOI was 1 h, 2 h, 4 h and 24 h. The transfection efficiency of Ad-EGFP were 0、89.29%、96.41%、96.72% and 98.37% respectively when MOI was 0, 50, 100, 200 and 500. The content of mIL-12 in the supernatant of AdmIL-12/H22 was (89.71±22.05) ng·48 h~(-1)·10~6 cells~(-1). AdmIL-12 and Ad-EGFP did not inhibit the growth of H22 cells. Conclusion:AdmIL-12 and Ad-EGFP were constructed successfully, and they can transfect H22 cells and express mIL-12 and EGFP respectively, but they do not inhibit the growth of H22 cells in vitro. That MOI is 100 and TOI is 2 hours, which are the best transfection condition of recombinant adenovirus on H22 cells.