Doki Y, Shiozaki H, Tahara H, Kobayashi K, Miyata M, Oka H, Iihara K, Mori T
Department of Surgery II, Osaka University Medical School, Japan.
Cancer. 1993 Sep 15;72(6):1813-8. doi: 10.1002/1097-0142(19930915)72:6<1813::aid-cncr2820720604>3.0.co;2-r.
The prognostic value of flow cytometric DNA analysis on paraffin-embedded tumor samples has been controversial in esophageal cancer. To clarify its true significance, the authors developed an improved method that excludes the possibility of contamination by lymphocytes in tumor sample.
Single nuclear suspension was prepared from paraffin-embedded samples on 103 patients with squamous cell carcinoma of the esophagus. Both DNA content and nuclear size were simultaneously measured by flow cytometry on 30,000 nuclei, and contaminated lymphocyte nuclei were eliminated from the data by optimal gating. Correlation between DNA ploidy and postoperative survival was examined.
Analysis using a flow cytometric cell sorter showed that the frequency of tumor cells in the lymphocyte-reducing gating fraction (LGF) was significantly higher than that in the conventional nongating fraction (NGF). LGF analysis showed aneuploid peaks in 58 patients (56.3%), but NGF analysis showed aneuploid peaks in only 38 patients. LGF analysis revealed that the aneuploid tumors had higher histologic grading (P < 0.05) and worse survival rate (P < 0.01) compared with diploid tumors. However, conventional methods could not detect this difference.
Flow cytometric analysis gating by nuclear size may be helpful to detect aneuploid peaks, and for predicting prognosis of patients with squamous cell carcinoma of esophagus.
流式细胞术对石蜡包埋肿瘤样本进行DNA分析在食管癌中的预后价值一直存在争议。为阐明其真正意义，作者开发了一种改进方法，排除肿瘤样本中淋巴细胞污染的可能性。
从103例食管鳞状细胞癌患者的石蜡包埋样本中制备单核悬液。通过流式细胞术对30000个细胞核同时测量DNA含量和核大小，并通过最佳设门从数据中剔除受污染的淋巴细胞核。检测DNA倍体与术后生存率之间的相关性。
使用流式细胞术细胞分选仪分析显示，淋巴细胞减少设门部分（LGF）中的肿瘤细胞频率显著高于传统的非设门部分（NGF）。LGF分析显示58例患者（56.3%）出现非整倍体峰，而NGF分析仅显示38例患者出现非整倍体峰。LGF分析显示，与二倍体肿瘤相比，非整倍体肿瘤具有更高的组织学分级（P < 0.05）和更差的生存率（P < 0.01）。然而，传统方法无法检测到这种差异。
通过核大小进行流式细胞术分析设门可能有助于检测非整倍体峰，并预测食管鳞状细胞癌患者的预后。
