[Preparation of anticolon carcinoma vaccine with rich chaperone peptides and study on its anticancer efficacy].
OBJECTIVE
To prepare the anticolon carcinoma vaccine with rich chaperone peptide and to examine its anticancer immunological efficacy.


METHODS
CT26 colon carcinoma cells were cultured in 1 mg/L Trichosanthin 1640 medium at different temperatures to induce the chaperone expression and promote the synthesis of antigen peptides. Groups of these cells treated under the different condition were lysed by the sonic disintegration, and the lysates were centrifuged. The rawpurified proteins were obtained from the supernatants by precipitating with saturated ammonium sulfate and removing the molecules below 50,000 and above 300,000 in molecular weight via dialysis. Furthermore, the proteins with the molecular weights in 70,000, 90,000, 95,000, 110,000 and 170,000 were collected through gel filtration and SDS-PAGE. The purified proteins were analysed by Western blotting, and inspected on the anticancer immunological effects including lymphocyte proliferation and the activities of NK and CTL.


RESULTS
Major of the chaperone peptides of anticancer effects in CT26 cells, including antigen peptides joining with HSP70, HSP90, gp96, HSP110 and HSP170, was satisfactorily extracted and condensed, and rich chaperone peptide composites were successfully obtained. The composites prepared under various condition could all enhance lymphocyte proliferation and the activities of CTL and NK(P<0.01).


CONCLUSIONS
The rich chaperone peptide composites are successfully prepared via dialysis, salt fractionation and gel filtration combined with SDS-PAGE. Both the heat stress and Trichosanthin can increase the composites, which treated by 42 centi-degree heat stress and Trichosanthin are found to possess the strongest anticancer efficacy.
Zhao
Jian-Gang
JG
Department of Colorectal Surgery, The Third People's Hospital of Hangzhou, Hangzhou 310009, China.
Huang
Chang-Xin
CX
Yang
Guan-Gen
GG
Jin
Jian-Fang
JF
Kang
Yun-Ping
YP
Xia
Da-Jing
DJ
Wang
Qing-Qing
QQ
D000818:Animals; D019496:Cancer Vaccines / Q000276:immunology*; D002478:Cells, Cultured; D018882:Heat Stress Disorders / Q000378:metabolism*; D006360:Heat-Shock Proteins / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D018832:Molecular Chaperones / Q000276:immunology*; D015978:Trichosanthin / Q000494:pharmacology
D019496:Cancer Vaccines; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; D015978:Trichosanthin