[The detection of Rb2/p130 and p53 gene mutations by denaturing high-performance liquid chromatography analysis of sputum of lung cancer patients and its clinical implications].
OBJECTIVE
To probe Rb2/p130 and p53 gene mutations at their hot-spots by denaturing high-performance liquid chromatography (DHPLC) analysis in sputum samples from lung cancer patients and to evaluate the feasibility of these gene markers in the clinical diagnosis of lung cancer.


METHODS
Genomic DNAs were extracted from 47 sputum samples (35 of lung cancer, 12 of benign lung disease) and their parallel peripheral blood lymphoid cells. The genomic DNAs were subjected to PCR amplification of Rb2/p130 gene at exon 19 - 22 and p53 gene at exon 5 - 9. The mutations of Rb2/p130 and p53 were detected by DHPLC by analysis of the PCR products.


RESULTS
Of the 47 sputum samples, the Rb2/p130 gene mutation detection rates were 22.86% (8/35) in the lung cancer group and 0 (0/12) in the control group (P = 0.049). The sensitivity and specificity were 22.86% and 100% respectively by using Rb2/p130 gene mutation detection as a diagnostic marker for lung cancer. p53 gene mutation detection rates were 28.57% (10/35) in the lung cancer group and 0 (0/12) in the control group (P = 0.046). The sensitivity and specificity were 28.57% and 100% respectively by using p53 gene mutation detection as a diagnostic marker for lung cancer. The sensitivity and specificity reached 51.43% and 100% respectively when Rb2/p130 and p53 gene mutations were combined as diagnostic markers for lung cancer.


CONCLUSIONS
Because of the low sensitivity, Rb2/p130 or p53 gene mutation detection alone is not feasible as a gene marker in the clinical diagnosis of lung cancer. The increased sensitivity, by combining the two gene markers, suggests that it may be feasible to use a panel of molecular markers such as p53, Rb2/p130, and others as diagnostic markers for lung cancer.
Department of Respiratory Disease, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing 100730, China.
Bin-chao
BC
Wang
Long-yun
LY
Li
Shu-jun
SJ
Cheng
Yan-ning
YN
Gao
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005240:Feasibility Studies; D005260:Female; D016158:Genes, p53; D006801:Humans; D008175:Lung Neoplasms / Q000175:diagnosis*; D008297:Male; D008875:Middle Aged; D009154:Mutation; D011506:Proteins / Q000235:genetics* / Q000378:metabolism; D050717:Retinoblastoma-Like Protein p130; D012680:Sensitivity and Specificity; D013183:Sputum / Q000378:metabolism*
D011506:Proteins; D050717:Retinoblastoma-Like Protein p130