流式细胞仪同时检测实体肿瘤P53蛋白和DNA含量

Simultaneous analysis of P53 expression and DNA content in solid cancer using flow cytometry

目的 :建立流式细胞仪检测方法 ,研究P5 3抑癌基因蛋白与DNA的关系。方法 :71例肿瘤手术样本 ,其中良性 8例 ,恶性 6 3例。①用机械方法获得实体肿瘤组织的单细胞悬液 ,1%多聚甲醛固定 ,或 5 0 %— 95 %乙醇固定 ,0 1%tritonX 10 0处理。先P5 3 FITC核抗原标记 ,再溴化丙啶 (PI)染DNA ,流式细胞仪双参数分析。②相同样本 32例P5 3蛋白免疫组化ABC法检测。结果 :1%多聚甲醛固定方法优于酒精固定法。流式细胞仪分析结果与免疫组化一致 (Kappa>0 0 4)。P5 3过量表达与DNA倍体相关 (P <0 0 0 5 )。结论 :流式细胞仪双参数检测DNA含量和癌基因蛋白是一个快速 。

Purpose:To study the relationship of P53 suppressor oncongene protein and DNA content in cancer cell and establish a simultaneous analysis of P53 expression and DNA contant by flow cytometry. Methods:We have studied the P53 protein in conjunction with DNA content in solid samples from 71 patients with tumor using flow cytometry. The suspension were obtained by mechanical disaggregation followed by a short fixation in 1% paraformadehyde. Immunohistochemical results obtained with the same monoclone antibody in the same 32 samples. Results:Paraformaldehyde fixed cells exhibited significantly greater positive ratio than ethanol fixed cells. The flow cytometric value correlated with immunohistochemical results. (Kappa>0 04). P53 over expression was significantly associated with DNA content ( P <0 005).Conclusions:Flow cytometry could be a feasible technique for clinical detection.