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Immunization with mutant HPV16 E7 protein inhibits the growth of TC-1 cells in tumor-bearing mice.

Li YL, Ma ZL, Zhao Y, Zhang J - Oncol Lett (2015)

Bottom Line: The expression of mE7 was induced by isopropyl β-D-1-thiogalactopyranoside.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.

View Article: PubMed Central - PubMed

Affiliation: School of Life Sciences, Shanghai University, Shanghai 200444, P.R. China ; Institute of Molecular Medicine, Nanjing University, Nanjing, Jiangsu 210093, P.R. China.

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 into Escherichia coli Rosetta. 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 2×10(5) TC-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.

No MeSH data available.


Related in: MedlinePlus

Inhibition of TC-1 growth by immunization with the mE7 protein. The mice were subcutaneously injected in the right flank with 1×105 TC-1 cells, and then immunized with the mE7 protein, supplemented with complete Freund’s adjuvant, the following day. PBS and E7 were used as controls. Each group contained 10 mice. Seven days later, mice were immunized using the same solution, supplemented with incomplete Freund’s adjuvant. (A) Tumor growth was monitored every three days for 24 days and (B) the survival rate was recorded. Tumor growth was determined by measuring the maximal and minimal diameters with a vernier caliper, and tumor volumes were calculated as follows: volume = (length × width2) × 0.52. PBS, phosphate-buffered saline; mE7, mutant non-transforming human papillomavirus 16 E7.
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f4-ol-09-04-1851: Inhibition of TC-1 growth by immunization with the mE7 protein. The mice were subcutaneously injected in the right flank with 1×105 TC-1 cells, and then immunized with the mE7 protein, supplemented with complete Freund’s adjuvant, the following day. PBS and E7 were used as controls. Each group contained 10 mice. Seven days later, mice were immunized using the same solution, supplemented with incomplete Freund’s adjuvant. (A) Tumor growth was monitored every three days for 24 days and (B) the survival rate was recorded. Tumor growth was determined by measuring the maximal and minimal diameters with a vernier caliper, and tumor volumes were calculated as follows: volume = (length × width2) × 0.52. PBS, phosphate-buffered saline; mE7, mutant non-transforming human papillomavirus 16 E7.

Mentions: Female C57BL/6 mice received an SC injection consisting of 1×105 TC-1 cells in the right flank, and were immunized with the mE7 protein, E7 protein or PBS in combination with CFA the following day. Seven days later, the mice were immunized using the same solution supplemented with IFA. Each group contained 10 mice. The TC-1 cell growth was significantly inhibited in the mice immunized with mE7 or E7compared with the control group (P<0.05; Fig. 4A). However, there was no significant difference in the survival rate of the mice immunized with mE7 and the control group (P=0.055), or E7 and the control group (P=0.105; Fig. 4B). No tumor metastasis was observed in any mouse.


Immunization with mutant HPV16 E7 protein inhibits the growth of TC-1 cells in tumor-bearing mice.

Li YL, Ma ZL, Zhao Y, Zhang J - Oncol Lett (2015)

Inhibition of TC-1 growth by immunization with the mE7 protein. The mice were subcutaneously injected in the right flank with 1×105 TC-1 cells, and then immunized with the mE7 protein, supplemented with complete Freund’s adjuvant, the following day. PBS and E7 were used as controls. Each group contained 10 mice. Seven days later, mice were immunized using the same solution, supplemented with incomplete Freund’s adjuvant. (A) Tumor growth was monitored every three days for 24 days and (B) the survival rate was recorded. Tumor growth was determined by measuring the maximal and minimal diameters with a vernier caliper, and tumor volumes were calculated as follows: volume = (length × width2) × 0.52. PBS, phosphate-buffered saline; mE7, mutant non-transforming human papillomavirus 16 E7.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4356387&req=5

f4-ol-09-04-1851: Inhibition of TC-1 growth by immunization with the mE7 protein. The mice were subcutaneously injected in the right flank with 1×105 TC-1 cells, and then immunized with the mE7 protein, supplemented with complete Freund’s adjuvant, the following day. PBS and E7 were used as controls. Each group contained 10 mice. Seven days later, mice were immunized using the same solution, supplemented with incomplete Freund’s adjuvant. (A) Tumor growth was monitored every three days for 24 days and (B) the survival rate was recorded. Tumor growth was determined by measuring the maximal and minimal diameters with a vernier caliper, and tumor volumes were calculated as follows: volume = (length × width2) × 0.52. PBS, phosphate-buffered saline; mE7, mutant non-transforming human papillomavirus 16 E7.
Mentions: Female C57BL/6 mice received an SC injection consisting of 1×105 TC-1 cells in the right flank, and were immunized with the mE7 protein, E7 protein or PBS in combination with CFA the following day. Seven days later, the mice were immunized using the same solution supplemented with IFA. Each group contained 10 mice. The TC-1 cell growth was significantly inhibited in the mice immunized with mE7 or E7compared with the control group (P<0.05; Fig. 4A). However, there was no significant difference in the survival rate of the mice immunized with mE7 and the control group (P=0.055), or E7 and the control group (P=0.105; Fig. 4B). No tumor metastasis was observed in any mouse.

Bottom Line: The expression of mE7 was induced by isopropyl β-D-1-thiogalactopyranoside.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.

View Article: PubMed Central - PubMed

Affiliation: School of Life Sciences, Shanghai University, Shanghai 200444, P.R. China ; Institute of Molecular Medicine, Nanjing University, Nanjing, Jiangsu 210093, P.R. China.

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 into Escherichia coli Rosetta. 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 2×10(5) TC-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.

No MeSH data available.


Related in: MedlinePlus