Evaluation of the chemosensitivity of head and neck cancer cells based on the diverse function of mutated-p53

Yasuhiro Shinagawa; Hitoshi Kawamata; Fumie Omotehara; Koh-Ichi Nakashiro; Mohammad O Hoque; Tadashi Furihata; Hideki Horiuchi; Yataka Imai; Takahiro Fujimori; Takashi Fujibayashi

Evaluation of the chemosensitivity of head and neck cancer cells based on the diverse function of mutated-p53

Int J Oncol. 2003 Feb;22(2):383-9.

Authors

Yasuhiro Shinagawa¹, Hitoshi Kawamata, Fumie Omotehara, Koh-Ichi Nakashiro, Mohammad O Hoque, Tadashi Furihata, Hideki Horiuchi, Yataka Imai, Takahiro Fujimori, Takashi Fujibayashi

Affiliation

¹ Department of Surgical and Molecular Pathology, Dokkyo University School of Medicine, Tochigi 321-0293, Japan.

PMID: 12527938

Abstract

Pre-therapeutic evaluation of p53 gene is very important for treating patients with head and neck cancer. However, the analysis for p53 gene has generally been done by immunohistochemistry, polymerase chain reaction (PCR)-single strand conformation polymorphism (SSCP) and direct sequencing. Functional analysis system for p53 transcriptional activity in mammalian cells is now required. We developed a functional analysis system for p53 transcriptional activity in cancer cells. We used two human head and neck cancer cell lines harboring mutated p53 gene, HSG (Asn30Ser) and TYS (Asp281His), and a human osteosarcoma cell line, Saos-2 as a control. We transfected these cells with luciferase reporter plasmids containing promoter sequence of p53 target genes (p21waf1, BAX, MDM2, p53AIP1 or PUMA). After treating the cells with chemotherapeutic drugs, alteration of the luciferase activity was measured. In HSG cells, none of the target gene promoters was activated by treatment with chemotherapeutic drugs. In TYS cells, p21waf1 promoter was markedly activated by treatment with chemotherapeutic drugs, but Bax and p53AIP1 promoter was not activated. This type of mutated-p53 in TYS cells prevents cell death from DNA damage, and probably accumulates genetic alterations and accelerates the malignant progression of the cells by DNA damaging therapy. Thus, analysis for the diverse function of mutated-p53 may help to determine the therapeutic strategy, especially for chemotherapy and radiation in the individual patients with head and neck cancer.

Publication types

Comparative Study
Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Substitution*
Antineoplastic Agents / adverse effects
Antineoplastic Agents / pharmacology*
Apoptosis
Apoptosis Regulatory Proteins
Carcinoma, Squamous Cell / genetics*
Carcinoma, Squamous Cell / pathology
Cell Transformation, Neoplastic / genetics*
Codon / genetics
Cyclin-Dependent Kinase Inhibitor p21
Cyclins / genetics
DNA Damage
DNA, Neoplasm / drug effects*
DNA, Neoplasm / genetics
Disease Progression
Drug Resistance, Neoplasm / genetics*
Genes, Reporter
Genes, p53*
Head and Neck Neoplasms / genetics*
Head and Neck Neoplasms / pathology
Humans
Luciferases / analysis
Luciferases / biosynthesis
Mutation, Missense*
Neoplasm Proteins / genetics
Neoplasm Proteins / physiology*
Nuclear Proteins*
Point Mutation*
Proteins / genetics
Proto-Oncogene Proteins / genetics
Proto-Oncogene Proteins c-bcl-2*
Proto-Oncogene Proteins c-mdm2
Recombinant Fusion Proteins / analysis
Recombinant Fusion Proteins / biosynthesis
Transfection
Tumor Cells, Cultured / pathology
Tumor Suppressor Protein p53 / genetics
Tumor Suppressor Protein p53 / physiology*
bcl-2-Associated X Protein

Substances

Antineoplastic Agents
Apoptosis Regulatory Proteins
BAX protein, human
BBC3 protein, human
CDKN1A protein, human
Codon
Cyclin-Dependent Kinase Inhibitor p21
Cyclins
DNA, Neoplasm
Neoplasm Proteins
Nuclear Proteins
P53AIP1 protein, human
Proteins
Proto-Oncogene Proteins
Proto-Oncogene Proteins c-bcl-2
Recombinant Fusion Proteins
Tumor Suppressor Protein p53
bcl-2-Associated X Protein
Luciferases
MDM2 protein, human
Proto-Oncogene Proteins c-mdm2