Objectives: To study the incidence and mechanism of the tumor suppressor gene p16 and RB inactivation in lung cancer, and to investigate their relations to biological characteristics, clinical pathological diagnosis and gene classification.
Methods: Immunohistochemistry, double in situ hybridization, PCR, PCR-SSCP, and sequencing were used to study the changes in the p16 and RB genes in 106 patients with lung cancer. The specimen of normal lung tissue from the patients with lung cancer and specimens of the 23 patients with benign lung diseases were studied in contrast to those of the same patients.
Results: The total expression rate of the p16 and RB genes in the lung cancer tissues of the 106 patients was lower than that in normal lung tissues and tissues of benign lung diseases and was specially related to histological type, metastasis, and clinical stage of lung cancer. In stage I and II lung cancer, the obvious inactivation rate of the tumor suppressor gene p16 or RB was 32.6% or 28.3%. The p16 inactivation appeared largely in non-small cell lung cancer (50.1%), and the RB inactivation largely in small cell lung cancer (88.2%). Homozygous deletion, methylation, and mutation were three mechanisms for the p16 gene inactivation. The deletion rate of the p16 exon 1 and/or exon 2 was 25.8% in the patients with non-small cell lung cancer. Fifteen patients (16.9%) showed methylation at the SmaI sites of CpG island of the p16 gene. PCR-SSCP and sequencing revealed p16 gene mutation in 9 of the 106 patients with lung cancer.
Conclusions: The p16 and RB genes may play an important role in genesis and progression of lung cancer. The inactivation of the p16 or RB gene may be an early sign of lung canceration, that is important for the early diagnosis. A new gene classification model for lung cancer diagnosis would be set up based on the research of the p16 and RB genes.