Preparation of TAT-apoptosis protein and its antitumor activity in vitro.

Objective To prepare TAT-apoptosis protein and study its antitumor activity. Methods The recombination plasmid pET-28b-TAT-vp3 was transferred into E. coli BL21(DE3) and the expressed inclusion body proteins were purif ied by Ni-NTA column chromatography. The purif ied protein was renaturated by dialysis and then the antitumor activity was studied. Results The inclusion body of target protein was expressed highly under 30 ℃ and the purity of TAT-apoptosis protein was above 92 % after the purif ication on Ni-NTA column with 500 mmol/L imidazole solution as the eluent. MTT test shown that the survival rate of HeLa cells was 42 % lower when TAT-apoptosis protein concentration was about 10 μg/mL (IC50=4.2 μg/mL). The animal experiments shown that TAT-apoptosis protein could inhibit the growth of tumor and the inhibition rate was about 48.3 %. Conclusion The purif ied TAT-apoptosis protein produced by E. coli has an antitumor effect by pharmacological assay.