定量RT-PCR检测穿孔素mRNA方法的建立与临床初步应用

Establishment and clinical application of quantitative RT-PCR method for detecting perforin mRNA

目的 建立检测穿孔素mRNA的定量RT PCR方法。方法 用限制性内切酶制备竞争性模板 ,建立定量RT PCR方法 ,检测正常人及部分肿瘤病人外周血的穿孔素mRNA水平。结果 6例正常人的外周血穿孔素mRNA含量为 0 5 1± 0 40pmol/ml,消化道肿瘤患者的穿孔素mRNA水平与正常相比 ,有显著差异 (P <0 .0 1) ,乳腺肿瘤及白血病患者的穿孔素mRNA水平与正常人相比 ,没有差异。结论 穿孔素mRNA水平的降低可能是肿瘤发生的一个重要原因 ;除穿孔素途径以外 ,还存在别的抗肿瘤的效应机制。

Objective\ To establish the quantitative method for detecting perforin mRNA. Methods \ The competitive templates were prepared by restriction endonuclease method.The quantitative RT PCR assay was established and used to detecting the perforin mRNA level in peripheral blood of tumor patients and healthy adults.Results\ Average perforin mRNA level of six healthy adults is 0.51±0.40 pmol/ml.The perforin mRNA level of patients with digestive tumors are significantly lower than that of healthy adults(P<0.01).But compared with healthy adults,no significant difference is showed in patients with breast cancer or leukemia.Conclusion\ The decreased perforin mRNA level probably attributes to the high incidence of cancers.Except for perforin mediated cytolysis,additional effective mechanism against tumor cells maybe exist.