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目的 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。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22例患者,其中确诊肺癌患者15例(涂片或活检找到癌细胞),临床高度怀疑肺癌患者5例(涂片或活检均阴性),2例非肿瘤患者(1例为结节病,另1为右中叶肺炎),同时采用正常肺组织标本作对照。结果 p53外显子6、7、8的PCRSSCP银染阳性率在肺癌组及癌疑组分别为6/15(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;PCRSSCP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 mutation in lung cancer patients and its significance in clinical diagnosis. Methods The Saccomanno method was used to enrich and preserve sputum specimens. Some of the smears were used to find cancer cells. The remaining specimens were subjected to DNA extraction of sputum cells and polymerase chain reaction (PCR) single strand conformation polymorphism analysis (SSCP). A total of 22 patients were examined. Among them, 15 were confirmed to have lung cancer (smear or biopsy to find cancer cells), 5 patients were clinically highly suspicious of lung cancer (negative smear or biopsy), and 2 were non-tumor patients (1 was sarcoidosis The other 1 was right middle lobe pneumonia. At the same time, normal lung tissue samples were used as controls. Results The positive rate of PCRSSCP silver staining in p53 exons 6, 7, and 8 was 6/15 (40%) and 2/5 (40%) in lung cancer group and cancer suspected group, respectively. Non-neoplastic patients had no p53 gene abnormalities in sputum specimens and normal lung tissues. Conclusion The modified Saccomanno method can not only store and condense technetium cells, but also facilitate the extraction of sputum specimen DNA. PCRSSCP silver staining method can be used to detect p53 gene mutation in sputum cells of lung cancer patients.