Yang He1, Bo Yang, Chang-Geng Ruan. 1. Jiangsu Provincial Institute of Hematology, The First Affiliated Hospital, Suzhou University, Suzhou, Jiangsu, 215006, P.R.China. heyang1963@163.com
Abstract
BACKGROUND & OBJECTIVE: Since most tumor antigens are uncertain, the immunotherapy against tumors still lingers in clinical trials. Because k-ras proto-oncogene specifically expresses in pancreatic cancer cells with constant mutation site, it may be an ideal target for immunotherapy against pancreatic cancer. This study was to evaluate the feasibility of inducing immunotherapy on pancreatic cancer by K-ras mutated peptides, give experimental evidence to clinical individual therapy on pancreatic cancer. METHODS: k-ras oncogene was amplified by reverse transcription-polymerase chain reaction (RT-PCR)u its mutation type was detected by flow cytometry. T cells were induced by dendritic cells, which had been pulsed with synthesized mutated peptide. Killing effect of tumor specific T cells on pancreatic cancer cell line Patu8988 was assessed by MTT assay. RESULTS: There was a point mutation (GGT-->GTT) in the 12th codon in Patu8988 cellu the amino acid was mutated to valine. The mutation epitope was efficiently presented on dendritic cells'surface. The cytotoxic T cells induced by mutated peptide could kill Patu8988 cells efficiently. CONCLUSIONS: The mutated peptide can efficiently induce immunocytes to kill pancreatic cancer cells, which have k-ras mutation site. This finding provides experimental clue for immunotherapy against pancreatic cancer.
BACKGROUND & OBJECTIVE: Since most tumor antigens are uncertain, the immunotherapy against tumors still lingers in clinical trials. Because k-ras proto-oncogene specifically expresses in pancreatic cancer cells with constant mutation site, it may be an ideal target for immunotherapy against pancreatic cancer. This study was to evaluate the feasibility of inducing immunotherapy on pancreatic cancer by K-ras mutated peptides, give experimental evidence to clinical individual therapy on pancreatic cancer. METHODS:k-ras oncogene was amplified by reverse transcription-polymerase chain reaction (RT-PCR)u its mutation type was detected by flow cytometry. T cells were induced by dendritic cells, which had been pulsed with synthesized mutated peptide. Killing effect of tumor specific T cells on pancreatic cancer cell line Patu8988 was assessed by MTT assay. RESULTS: There was a point mutation (GGT-->GTT) in the 12th codon in Patu8988 cellu the amino acid was mutated to valine. The mutation epitope was efficiently presented on dendritic cells'surface. The cytotoxic T cells induced by mutated peptide could kill Patu8988 cells efficiently. CONCLUSIONS: The mutated peptide can efficiently induce immunocytes to kill pancreatic cancer cells, which have k-ras mutation site. This finding provides experimental clue for immunotherapy against pancreatic cancer.