title: Immunization with mutant HPV16 E7 protein inhibits the growth of TC-1 cells in tumor-bearing mice.
abstract: Two human papillomavirus (HPV) 16 oncogenic proteins, E6 and E7, are co-expressed in the majority of HPV16-induced cervical cancer cells. Thus, the E6 and E7 proteins are good targets for developing therapeutic vaccines for cervical cancer. In the present study, immunization with the mutant non-transforming HPV16 E7 (mE7) protein was demonstrated to inhibit the growth of TC-1 cells in the TC-1 mouse model. The HPV16 mE7 gene was amplified by splicing overlap extension polymerase chain reaction using pET-28a(+)-E7 as a template, and the gene was cloned into pET-28a(+) to form pET-28a(+)-mE7. Compared with the E7 protein, mE7 lacks amino acid residues 94-98, and at residue 24, there is a Cys to Gly substitution. pET-28a(+)-mE7 was then introduced intoEscherichia coliRosetta. The expression of mE7 was induced by isopropyl β-D-1-thiogalactopyranoside. The mE7 protein was purified using Ni-NTA agarose and detected by SDS-PAGE and western blot analysis. In the tumor prevention model, no tumor was detected in the mice vaccinated with the mE7 protein. After 40 days, the tumor-free mice and control mice were challenged with 2×105TC-1 cells. All control mice developed tumors six days later, but mE7 immunized mice were tumor free until 90 days. In the tumor therapy model, the TC-1 cells were initially injected subcutaneously, and the mice were subsequently vaccinated. Vaccination against the mE7 protein may significantly inhibit TC-1 cell growth compared to the control. These results demonstrated that immunization with the HPV16 mE7 protein elicited a long-term protective immunity against TC-1 tumor growth and generated a significant inhibition of TC-1 growth in a TC-1 mouse model.
authors: School of Life Sciences, Shanghai University, Shanghai 200444, P.R. China ; Institute of Molecular Medicine, Nanjing University, Nanjing, Jiangsu 210093, P.R. China.
authors: Yan-Li
authors: YL
authors: Li
authors: School of Life Sciences, Shanghai University, Shanghai 200444, P.R. China.
authors: Zhong-Liang
authors: ZL
authors: Ma
authors: Institute of Molecular Medicine, Nanjing University, Nanjing, Jiangsu 210093, P.R. China.
authors: Yue
authors: Y
authors: Zhao
authors: Institute of Molecular Medicine, Nanjing University, Nanjing, Jiangsu 210093, P.R. China.
authors: Jing
authors: J
authors: Zhang
keywords: TC-1 cells; human papillomavirus; immunization; mutant E7 protein