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目的 :为了解痰标本检测肺癌患者 P53 基因突变的可行性及其对临床诊断的意义。材料与方法 :采用改良 Saccomanno法浓缩及保存痰标本 ,部分涂片找癌细胞 ,其余标本进行痰细胞 DNA提取及多聚酶链反应 ( PCR) -单链构象多态性分析 ( SSCP) ,共检测 2 2例病人 ,其中确诊肺癌病人 1 5例 (涂片或活检找到癌细胞 )、临床高度怀疑肺癌病人 5例 (涂片或活检均阴性 )、2例非肿瘤病人 ( 1例为结节病 ,另 1为左中叶肺炎 ) ,同时采用正常肺组织标本作对照。结果 :P53 外显子 6,7,8的 PCR-SSCP银染阳性率在肺癌组及癌疑组分别为 6/ 1 5 (40 % )和 2 / 5 (40 % )。非肿瘤患者痰标本及正常肺组织均未发现 P53基因异常。结论 :改良 Saccomanno法不仅能贮存、浓缩痰细胞 ,而且方便痰标本 DNA提取 ;PCR-SSCP银染方法可用来检测肺癌患者痰细胞的 P53基因突变。
Objective: To understand the feasibility of sputum specimens detecting P53 gene mutations in patients with lung cancer and its clinical significance. MATERIALS AND METHODS: Saccomanno method was used to concentrate and preserve sputum specimens. Some smears were taken for cancer cells. The remaining specimens were used for DNA extraction and PCR-SSCP analysis. Two patients were diagnosed as lung cancer (including smear or biopsy), five were highly suspected lung cancer patients (both smear and biopsy were negative), two were non-tumor patients (one was sarcoidosis, The other for the left middle lobe pneumonia), while using normal lung tissue samples for the control. Results: The positive rates of PCR-SSCP silver staining of P53 exons 6, 7 and 8 were 6/1 5 (40%) and 2/5 (40%) in the lung cancer group and the cancer-suspected group respectively. Non-tumor patients sputum specimens and normal lung tissue were not found P53 gene abnormalities. Conclusion: The modified Saccomanno method can not only store and concentrate sputum cells, but also facilitate DNA extraction from sputum samples. The PCR-SSCP silver staining method can be used to detect the mutation of P53 gene in sputum cells of lung cancer patients.