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目的利用微卫星多态性分析探寻非小细胞肺癌(NSCLC)术后肺内复发转移与第二原发肿瘤(Second primary tumor,SPT)的分子遗传学差异以助鉴别诊断。方法 1994年1月至2002年8月接受手术治疗的21例Ⅰ～Ⅲ_A 期 NSCLC 患者,术后2～7年(中位2.8年)发现病理类型与原发 NSCLC 相同的肺内孤立肿瘤(腺癌7例,鳞癌14例),将之与相应的原发 NSCLC 石蜡包埋组织配对,以非恶性肺组织为对照,在显微组织切割基础上分离其 DNA,应用 PCR 为基础的微卫星分析方法检测8个定位于染色体3p、9p 及17p 区域的高密度微卫星位点,确定每一位点在相应配对肿瘤组织中杂合性丢失的异同,并结合临床特点分析肺内孤立肿瘤是复发转移还是 SPT。结果根据临床及影像学特点,9例患者肺内孤立肿瘤转移的可能性较大,其中7例配对肿瘤组织在3p、9p 及17p 8个微卫星标志上等位基因杂合性丢失的位点相同,提示肺内孤立肿瘤与原发 NSCLC 具有共同的克隆起源,支持肺内转移癌的诊断。4例肺内孤立肿瘤患者的临床病理及影像学特点倾向 SPT,其配对肿瘤3p、9p 及17p 上8个微卫星位点等位基因杂合性丢失条带不一致,表明肺内孤立肿瘤克隆起源相异于原发 NSCLC,支持临床 SPT 的诊断。另8例肺内孤立肿瘤患者术前不能判定系 SPT 还是肺内转移,其中6例配对组织3p 或3p 与17p 微卫星位点杂合性丢失模式一致,但9p 或17p 杂合性丢失不一致,临床随访发现该6例肺内孤立肿瘤术后6～13个月内可见肺内或远处转移,支持肺内孤立肿瘤为转移癌的诊断;2例配对肿瘤虽有一致的9p 或17p 微卫星位点杂合性丢失,但3p 的杂合性丢失模式不一致,临床倾向 SPT。结论对 NSCLC 术后肺内孤立病灶,3p、9p、17p 微卫星多态性分析其等位基因杂合性丢失有助于鉴别系肺内转移还是 SPT,其中3p 的杂合性丢失可能是鉴别诊断的重要生物学标志,需扩大样本进一步研究。
Objective To explore the molecular genetic differences between intrapulmonary recurrent metastasis and second primary tumor (SPT) in non-small cell lung cancer (NSCLC) after microsatellite polymorphism analysis to help differential diagnosis. Methods Twenty-one patients with stage Ⅰ-Ⅲ_A NSCLC who underwent surgery from January 1994 to August 2002 were enrolled in this study. Isolated lung tumor (adenocarcinoma) of the same pathological type and primary NSCLC was found 2 to 7 years (median 2.8 years) 7 cases of cancer and 14 cases of squamous cell carcinoma), matched with the corresponding primary NSCLC paraffin-embedded tissues, non-malignant lung tissue as a control, DNA isolation on the basis of microscopic cutting, and application of PCR-based microsatellite Analysis of eight high-density microsatellite loci located on the chromosomes 3p, 9p and 17p to determine the loss of heterozygosity for each locus in the corresponding matched tumor tissue, and analysis of clinical characteristics of isolated lung tumor is Recurrence and metastasis or SPT. Results According to the clinical and radiological features, the possibility of isolated tumor metastasis in the lungs of 9 patients was high. Among the 7 matched pairs of tumor tissues, there were 8 alleles at the 3p, 9p and 17p microsatellite loci The same, suggesting that isolated lung tumor with primary NSCLC have a common origin of cloning, support for the diagnosis of metastatic lung cancer. The pathological and imaging features of 4 patients with solitary pulmonary disease were not consistent with the loss of heterozygosity of the 8 microsatellite loci on 3p, 9p and 17p, indicating that the origin of isolated lung tumor clones Different from the primary NSCLC, to support the diagnosis of clinical SPT. In the other 8 cases of isolated solitary pulmonary tumor, we could not determine whether SPT or intrapulmonary metastasis were preoperatively. The pattern of loss of heterozygosity of 3p or 3p in 6 paired tissues was the same as that of 17p microsatellite locus, but the loss of heterozygosity of 9p or 17p was inconsistent, Clinical follow-up found that 6 cases of solitary pulmonary tumor within 6 to 13 months after operation can be seen in the lung or distant metastasis, support for the diagnosis of isolated lung cancer metastasis; 2 matched paired tumor despite the same 9p or 17p microsatellite Site heterozygosity is lost, but 3p heterozygosity loss patterns are inconsistent, clinical tendency SPT. Conclusion It is helpful to identify the loss of heterozygosity of 3p, 9p, 17p microsatellite polymorphism in intrapulmonary isolated lesions of NSCLC in differentiating intrapulmonary metastasis or SPT. The loss of heterozygosity of 3p may be identified Important biomarkers for diagnosis need to be scaled up for further study.