title: Recombinant expression of different mutant K-ras gene in pancreatic cancer Bxpc-3 cells and its effects on chemotherapy sensitivity.
abstract: K-ras is a member of ras gene family which is involved in cell survival, proliferation and differentiation. When a mutation occurs in ras gene, the activation of Ras proteins may be prolonged to induce oncogenesis. However, the relationship between K-ras mutation and clinical outcomes in pancreatic cancer patients treated with chemotherapy agents is still under debate. In this study, we constructed five pAcGFP1-C3 plasmids for different types of K-ras gene (WT, G12V, G12R, G12D, and G13D) and stably transfected human pancreatic cancer Bxpc-3 cells with these genes. The wild type and mutant clones showed a comparable growth and expression of K-Ras-GFP fusion protein. The expression of some K-ras mutations resulted in a reduced sensitivity to gefitinib, 5-FU, docetaxel and gemcitabine, while showed no effects on erlotinib or cisplatin. Moreover, compared with the wild type clone, K-Ras downstream signals (phospho-Akt and/or phospho-Erk) were increased in K-ras mutant clones. Interestingly, different types of K-ras mutation had non-identical K-Ras downstream signal activities and drug responses. Our results are the first to reveal the relationship between different K-ras mutation and drug sensitivities of these anti-cancer drugs in pancreatic cancer cells in vitro.
authors.affiliation: Department of Clinical Pharmacy, School of Pharmacy, Fudan University, Shanghai, 201203, China.
authors.forename: TengFei
authors.initials: T
authors.lastname: Shao
authors.forename: YuanTing
authors.initials: Y
authors.lastname: Zheng
authors.forename: Bei
authors.initials: B
authors.lastname: Zhao
authors.forename: Tao
authors.initials: T
authors.lastname: Li
authors.forename: KeGuang
authors.initials: K
authors.lastname: Cheng
authors.forename: WeiMin
authors.initials: W
authors.lastname: Cai
mesh_terms: D000970:Antineoplastic Agents / Q000737:chemistry / Q000627:therapeutic use*; D017209:Apoptosis; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002470:Cell Survival; D002945:Cisplatin / Q000627:therapeutic use; D004354:Drug Screening Assays, Antitumor; D000069347:Erlotinib Hydrochloride; D005472:Fluorouracil / Q000627:therapeutic use; D000077156:Gefitinib; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D011905:Genes, ras; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D009154:Mutation; D010179:Pancreas / Q000378:metabolism; D010190:Pancreatic Neoplasms / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D011799:Quinazolines / Q000627:therapeutic use; D011994:Recombinant Proteins / Q000737:chemistry; D018631:ras Proteins / Q000378:metabolism
chemical_list: D000970:Antineoplastic Agents; D011799:Quinazolines; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D000069347:Erlotinib Hydrochloride; D018631:ras Proteins; D002945:Cisplatin; D000077156:Gefitinib; D005472:Fluorouracil