摘要
[目的]
1 DHPLC分析肺癌患者组织标本Rb2/p130和p53突变。
2 DHPLC分析肺癌患者痰标本Rb2/p130和p53突变及其作为临床分子诊断标记的可行性。
[方法]
1 基因组DNA提取自52例肺癌组织标本(44例NSCLC,8例SCLC)和47例痰标本(35例肺癌,12例对照)以及相应的外周血淋巴细胞。
2 PCR扩增Rb2/p130基因Exon19~22和p53基因Exon5~9。
3 DHPLC分析PCR扩增产物检测Rb2/p130和p53基因突变。
[结果]
1 组织标本Rb2/p130突变检出率23.08%(12/52),其中腺癌32.00%(8/25)、鳞癌15.79%(3/19)、小细胞癌12.50%(1/8),三者差异无显著意义(P=0.403)。Exon21突变10例次,Exon20和Exon22突变各1例,未见Exon19突变。44例NSCLC Rb2/p130突变检出率与患者的性别、年龄、吸烟、病理类型、疾病分期等均无相关性(所有P>0.05)。痰标本肺癌组Rb2/p130突变检出率22.86%(8/35),对照组未检出一例,差异有显著意义(P=0.049)。痰标本检测Rb2/p130基因突变作为肺癌诊断分子标记,特异度为100%,灵敏度为22.86%。
2 组织标本p53突变检出率43.23%(23/52),其中NSCLC 40.91%(18/44),SCLC 62.50%(5/8),两者差异无显著意义(P=0.067)。Exon5~6检出10例次、Exon7突变6例次、Exon8~9突变9例次,一例同时并Exon5~6和Exon8~9突变,另一例同时并Exon7和Exon8~9突变。44例NSCLC p53突变检出率与患者的性别、吸烟、病理类型、疾病分期均无相关性(P>0.05),但在高龄(≥60岁)患者检出率高于低龄(<60岁)患者(P=0.01)。痰标本肺癌组p53突变检出率28.57%(10/35),对照组未检出一例,差异有显著
Objectives
1. 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.
Methods
1. 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.
Results
1. 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.
引文
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