CK19 RT-PCR法检测淋巴结中肺癌微小转移灶

Detection of lung cancer micrometastasis in regional lymph nodes by assays of CK19 reverse transcription polymerase chain reaction

目的 探索检测淋巴结中肺癌微小转移灶的新途径。方法 区域性淋巴结共 15 9枚 ,取自 2 0例可手术的原发性肺癌患者 ,将每枚淋巴结均分为两等份 ,分别进行癌转移的病理学检测和细胞骨架角蛋白 19(CK19)基因的表达分析。结果 本实验建立的CK19RT PCR法可检测到 1× 10 7个正常淋巴细胞中存在的 10个肺癌细胞。在被测的 15 9枚淋巴结中 ,42枚被病理切片和分子学方法同时证实存在转移 ,在余下的 117枚病检阴性淋巴结中 ,CK19RT PCR还发现 2 5枚存在微小转移。结论 与传统病理学检查相比 ,CK19RT PCR法可提高淋巴结癌转移检出率 。

Objective To set up a molecular method(RT PCR) which can be used to detect micrometastasis of regional lymph nodes(LNs) in patients with lung cancer. Methods Primary lung cancer tissues (n=20) and regional LNs (n=159) were obtained from 20 patients with lung cancer who underwent lobectomy. Each LN was halved. One half of a LN was subjected to histological examination (HE) and the other half was subjected to RT PCR amplification of CK19 mRNA. Serial dilution study using LC 5 cells was performed to detect sensitivity of the CK19 RT PCR method. Results Serial dilution study for LC 5 cells demonstrated that CK19 mRNA was detectable at a concentration as low as 10 LC 5 cells in 1×10 7 LN cells. CK19 mRNA was found in 42 LNs that were proved to have metastasis by HE. Of the other 117 LNs which were diagnosed as no metastasis by HE, however, twenty five LNs were found to express CK19 mRNA by RT PCR.Conclusion Comparing with HE method, the RT PCR method can make more accurate assessment of metastatic status in LNs.