流式细胞仪同时检测实体肿瘤P53蛋白和DNA含量

目的：建立流式细胞仪检测方法，研究Ｐ５３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６３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５０％～９５％乙醇固定，０．１％ｔｒｉｔｏｎ Ｘ－１００处理，先Ｐ５３－ＦＴＴＣ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Ｐ５３蛋白免疫组化ＡＢＣ法检测。结果：１％多聚甲醛

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

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.