摘要
目的完善流式细胞术DNA指数计算方法,准确定量分析肿瘤细胞DNA含量,为临床恶性肿瘤的早期诊断、治疗及预后分析提供可靠的技术方法。方法利用流式细胞术定量分析技术,在肿瘤单细胞悬液内加入20%鸡血红细胞和同型正常组织细胞。设立新的DNA指数(DNA Index,DI)。计算公式,DI=样品细胞G_0/G_1峰均道值/鸡血红细胞均道值×比值(指鸡血红细胞与实验样品同型对照正常细胞G_0/G_1期均道值之比)。依据此公式计算DNA指数。同时与传统方法进行比较,传统方法公式为,DI=样品细胞G_0/G_1峰均道值/正常细胞G_0/G_1峰均道值。结果食管癌40例DI值为1.75±0.64,增值指数(proliferation Index,PI)为0.58±0.14,DNA异倍体率为92.50%。贲门癌组为30例DI值为1.54±0.27,PI值为0.58±0.12,DNA异倍体率为93.40%。胃癌组30例,DI值为1.50±0.26,PI值为0.56±0.15,DNA异倍体率为90.00%。正常组40例DI值为1.03±0.03,PI值为0.19±0.05,无DNA异倍体出现。3种恶性肿瘤利用传统方法检测的DNA异倍体率为82.4%,双内标法检测的DNA异倍体率为92.0%。各组间经统计学处理差异有统计学意义(P<0.01)。结论双内标法有其一定的优越性,可以消除来自各方面的干扰因素对DNA染色和荧光强度的影响,更加准确地反映肿瘤DNA倍体与同型组织细胞DNA倍体的关系,对肿瘤的诊断病理分级及分型、预后判断和治疗效果均有指导意义。
Objective To improve the quantitative analysis of DNA ploid in tumor cells detected by flow cytometry, which could provide diagnosis, treatment and prognosis analysis methods for early malignant tumor in clinic. Methods To put 20% chicken red blood cells and homotype normal cells into tumor cells,then DNA ploid was detected by flow cytometry. New DNA Index （DI） was set, DI = Go/G1 mean fluorescence intensity of tumor cells/G0/G1 mean fluorescence intensity of chicken red blood cells x ratio, ratio = G0/G1 mean fluorescence intensity of chicken red blood cells/G0/G1 mean fluorescence intensity of homotype normal cells. New DI method was compared with traditional DI method. Traditional DI = G0/G1 mean fluorescence intensity of tumor cells/G0/G1 mean fluorescence intensity of normal cells. Results In 40 cases esophageal cancer, DI was 1.75 ± 0.06, proliferation Index （PI） was 0.58 ± 0.14, DNA heteroploidy rate was 92.50%. In 30 cases carcinoma of gastric cardia, DI was 1.54 ± 0.27, PI was 0.58 ±0. 12 ,DNA heteroploidy rate was 93.40%. In 30 cases gastric carcinoma, DI was 1.50 ± 0.26, PI was 0.56 ± 0.15. In 40 cases normal tissuses, DI was 1.03 ± 0.03, PI was 0.19 ± 0.05, and no DNA heteroploidy. DNA heteroploidy rate was 82.4% with traditional method, DNA heteroploidy rate was 92.0% with new method. Conclusion DI detected by double internal standards （ chicken red blood cells and homotype normal cells） was better than traditional method with high accuracy and low error, which eould eliminate confounding factors that could influnce the DNA staining and fluorescence intensity. The new method could exactly reflect the relationship between DNA ploid of tumor and DNA ploid of homotypetissues, and have guidance significance for oneopathologic grading and typing, and evaluating prognosis and therapeutic efficacy in clinic.
出处
《河北医科大学学报》
CAS
2010年第5期558-561,共4页
Journal of Hebei Medical University
基金
河北省普通高等学校强势特色学科建设资助项目(冀教高[2005]52号)
河北省科技计划项目资助(07276101D-99)
关键词
食管肿瘤
胃肿瘤
流式细胞术
esophageal neoplasms
stomach neoplasms
flow cytoctmetry
