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目的探讨应用荧光定量聚合酶链反应 (FQ PCR)扩增乳腺组织特异性基因hMAMmRNA作为检测乳腺癌血行微小转移方法的可能性。方法利用逆转录RT PCR检测hMAMmRNA在乳腺癌细胞系SKBR3的表达 ,将FQ PCR与普通PCR的敏感性进行比较 ,并利用FQ PCR对乳腺癌等不同病种患者 10 1例及健康志愿者 31例的外周血进行检测 ,分析hMAMmRNA表达与乳腺癌临床特点之间的关系。结果普通巢式PCR扩增hMAMmRNA在 10 6个白细胞中可检测出 1个SKBR3细胞 ,而巢式FQ PCR可在 10 7个白细胞中可检测出 1个SKBR3细胞 ,6 3例乳腺癌患者外周血中 19例hMAMmRNA阳性 (阳性率 30 % ) ,随肿瘤分期进展阳性率增高。其他病种患者外周血中均阴性 ,而 31例健康献血员中有 1例阳性。结论巢式FQ PCR是检测乳腺癌微小转移的敏感方法
Objective To investigate the possibility of using fluorescence quantitative polymerase chain reaction (FQ PCR) to amplify breast tissue-specific gene hMAMmRNA as a micrometastasis detection method in breast cancer. Methods Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the expression of hMAM mRNA in breast cancer cell line SKBR3. The sensitivity of FQ PCR and ordinary PCR was compared. FQ PCR was used to detect the expression of hMAM mRNA in 101 different breast cancer patients and 31 healthy volunteers Of peripheral blood were detected and analyzed the relationship between hMAM mRNA expression and clinical features of breast cancer. Results Normal nested PCR amplification of hMAM mRNA detected 1 SKBR3 cell in 10 6 white blood cells, while nested FQ PCR detected 1 SKBR3 cell in 10 7 white blood cells. The peripheral blood of 63 breast cancer patients In 19 cases of hMAM mRNA positive (positive rate 30%), with the staging of the tumor positive rate increased. Other disease patients were negative in the peripheral blood, and 31 healthy blood donors in 1 positive. Conclusion Nested FQ PCR is a sensitive method to detect micrometastases in breast cancer