[Detection of multiple gene mutations in non-small-cell lung cancer by suspension microarray].
OBJECTIVE
To construct a high-throughput suspension microarray for detecting the hotspot gene mutations of p53, p16, retinoblastoma (Rb) and epidermal growth factor receptor (EGFR) and to investigate the significance of this multimarker panel in molecular diagnosis of non-small-cell lung cancer (NSCLC).


METHODS
The specific probes of normal or mutated sequences targeting the hotspot mutation sites of p53, p16, Rb and EGFR were designed and immobilized to carboxylated Luminex microspheres (micro-beads). Genomic DNA was extracted from 65 specimens of cancer tissues and 20 adjacent normal lung tissues. p53, p16, Rb and EGFR genes were amplified by PCR, hybridized with the specific probes on the beads and measured using Luminex 100.


RESULTS
The single gene mutations of p53, p16, Rb or EGFR in NSCLC specimens were 53.8% (35/65), 20.0% (13/65), 7.7% (5/65) or 35.4% (23/65) respectively. The para-tumor normal tissue specimens were 5.0% (1/20), 5.0%(1/20), 0 and 0 respectively. For combined detections of four genes, the sensitivity, specificity and accuracy were 81.5% (53/65), 90.0% (18/20) and 83.5%(71/85) respectively. The mutation rates of this panel in stage I, stage II and stage III were 78.3% (18/23), 80.0% (16/20) and 86.4% (19/22) respectively.


CONCLUSIONS
A high-throughput suspension microarray with a higher specificity and sensitivity has been built. It may be used to simultaneously detect the gene mutations of p53, p16, Rb or EGFR in NSCLC specimens. This suspension microarray is helpful to improve the sensitivity of molecular diagnosis of NSCLC and guide the molecular targeting therapy of NSCLC.
Wang
Dong
D
Cancer Center of PLA, 81st Hospital of PLA, Nanjing 210002, China. wangdong_nj001@126.com
Ye
Yu-kun
YK
Su
Chang-qing
CQ
Rong
Tie-hua
TH
Guo
Ai-yong
AY
Yi
Jin-hua
JH
Liu
Yan-jun
YJ
D000293:Adolescent; D000328:Adult; D000368:Aged; D014408:Biomarkers, Tumor; D002289:Carcinoma, Non-Small-Cell Lung / Q000175:diagnosis* / Q000235:genetics*; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000235:genetics; D004252:DNA Mutational Analysis; D066246:ErbB Receptors / Q000235:genetics; D005260:Female; D016158:Genes, p53; D006801:Humans; D008175:Lung Neoplasms / Q000175:diagnosis* / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis; D016160:Retinoblastoma Protein / Q000235:genetics; D012680:Sensitivity and Specificity; D016159:Tumor Suppressor Protein p53 / Q000235:genetics; D055815:Young Adult
D014408:Biomarkers, Tumor; D019941:Cyclin-Dependent Kinase Inhibitor p16; D016160:Retinoblastoma Protein; D016159:Tumor Suppressor Protein p53; D066246:ErbB Receptors