原位逆转录PCR检测干细胞因子mRNA在白血病细胞的表达

Detection of stem cell factor mRNA expression in leukemic cells by in situ reverse transcriptase-PCR

目的探索应用原位逆转录PCR(RT-PCR)检测低拷贝的干细胞因子mRNA在白血病细胞表达的可行性。方法在普通的PE480型PCR仪进行原位RT-PCR,检测干细胞因子mRNA在K562、HL-60和慢性粒细胞性白血病细胞的表达。结果直接法和间接法原位RT-PCR均可以检测到干细胞因子mRNA在K562、HL-60和慢性粒细胞性白血病细胞表达,而常规的原位杂交则为阴性。结论(1)干细胞因子在K562、HL-60和慢性粒细胞性白血病细胞均有表达,但是表达量较低,应用原位RT-PCR可进行检测;(2)在普通的PE480型PCR仪上可以进行原位PCR。

Objective To evaluate the feasibility of detecting low copy stem cell factor (SCF) mRNA in leukemic cells by in situ reverse transcriptase-PCR (RT-PCR). Methods In situ RT-PCR was carried out on PE480 DNA Thermal Cycler to determine the expression of SCF mRNA in K562, HL-60 and chronic myelogenous leukemia (CML) cells in blast phase. Results SCF mRNA was detected in the 3 cell lines by direct or indirect in situ RT-PCR, but not by in situ hybridization. Conclusion SCF mRNA is present in K562, HL-60 and CML cells in blast phase but with very low expression level which can be detected by in situ RT-PCR carried out on PE480 DNA Thermal Cycler.