Detection Of Rb2/p130 And P53 Gene Mutations By DHPLC Analysis In Chinese Lung Cancer Patients And Its Clinical Implications

Objectives1. To probe Rb2/p130 and p53 gene mutations at their hot-spots by DHPLC analysis in Chinese lung cancer patients.2. To evaluate the feasibility of the two gene markers as a clinical diagnostic avenue of lung cancer.Methods1. Genomic DNAs were extracted from 52 biopsy samples (44 of NSCLC, 8 of SCLC), 47 sputum samples (35 of lung cancer, 12 of benign lung disease), and their parallel peripheral blood lymphoid cells.2. To subject the genomic DNAs to PCR amplification of Rb2/p130 gene at exon 19-22 and p53 gene at exon 5-9.3. The mutations of Rb2/p130 and p53 were detected by DHPLC analyzing the PCR products.Results1. 52 biopsy samples with a total detection rate of 23.08%( 12/52), in which 32.00%(8/25) of adenocarcinoma, 15.79%(3/19) of squamous carcinoma, and 12.50%(l/8) of small cell carcinoma, for Rb2/p130 gene mutations were confirmed. Ten cases were detected mutations at exon 21, one at exon 20, one at exon 22, but none at exon 19. No relations were found between the total detection rate of Rb2/p130 gene mutation and the patients' sexes, ages, stages, smoking, and pathologies in 44 NSCLC (all P>0.05). Of 47 sputum samples, the Rb2/p130 gene mutation detection rates were 22.86%(8/35) in lung cancer group and 0%(0/12) in control group (P=0.049). The sensitivity and specificity were 22.86% and 100% respectively by using Rb2/p130 gene mutation detected in sputum sample as a diagnostic marker for lung cancer.