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目的 以 RT- PCR检测胃周淋巴结胃癌转移细胞并评价其临床应用价值。 方法 以 MUC1c DNA的特异序列为 RT- PCR引物 ,酶切分析法及系列稀释法对该法的特异性和敏感性进行分析 ,对临床收集的胃周淋巴结样品作 RT- PCR检测及产物 DNA点杂交验证。 结果 ( 1)胃及胃癌组织均存在 MUC1m RNA的表达 ;( 2 )该扩增体系具有较好的特异性 ;( 3)敏感性可达 1pgRNA,相当于从 10 5个淋巴细胞中检出 1个胃癌细胞 ;( 4)对临床样品检测的结果显示较病理检查敏感 ,PCR产物点杂交进一步证实 MUC1作为 PCR标志物有较好的可靠性。 结论 该法具有较高的可靠性和较好的应用前景。
Objective To detect the lymph node metastasis of gastric cancer by RT-PCR and evaluate its clinical value. Methods The specificity and sensitivity of the specific sequence of MUC1c DNA as RT-PCR primers, enzyme digestion analysis and serial dilution method were analyzed. RT-PCR detection and product DNA spots were performed on clinically collected lymph node samples. Cross validation. Results (1) Expression of MUC1m RNA was found in stomach and gastric cancer tissues; (2) The amplification system had better specificity; (3) The sensitivity was up to 1 pgRNA, which was equivalent to that detected from 10 5 lymphocytes. 1 One gastric cancer cell; (4) The results of clinical sample detection were more sensitive than pathological examination. PCR product spot hybridization further confirmed MUC1 as a PCR marker has better reliability. Conclusion This method has high reliability and good application prospects.