外周血P糖蛋白功能检测及其与乳腺癌多药耐药性的关系

目的 建立人外周血NK细胞P糖蛋白功能检测的方法,分析P糖蛋白功能与乳腺癌患者原发性多药耐药之间的关系.方法 选择经含蒽环类和紫杉类药物联合化疗的初治乳腺癌患者16例,治疗后化疗敏感和耐药者各8例.采集患者外周血标本,分离NK细胞,与荧光物质罗丹明123(Rh123)孵育,应用流式细胞仪检测不同时间点外周血NK细胞内Rh123的荧光强度;以Rh123的荧光强度(F)和相应时间点(t)作图,通过曲线估计回归分析,构建P糖蛋白药泵功能检测的最佳数学模型,计算每例患者相应的外排速率常数;分析化疗敏感组和耐药组患者速率常数的差异,以及以速率常数预测乳腺癌原发性多药耐药的可行性.结果 所有患者的NK细胞在撤离Rh123后60min,基本不再外排Rh123,NK细胞内Rh123的蓄积量、排出量和残留量均与化疗敏感性之间无明显相关性.通过回归分析建立了P糖蛋白功能检测的最佳数学模型F1=F0·e-kt(F1为t时间点的荧光强度,k为外排速率常数),化疗敏感组与耐药组患者外排速率常数差异有统计学意义(P=0.025).以k>3.9为标准,诊断乳腺癌原发性多药耐药,其诊断敏感性为75.0%,特异性为100%.准确率87.5%.结论 外周血NK细胞P糖蛋白药泵外排功能与乳腺癌患者原发性多药耐药密切相关,外排速率常数可能是预测乳腺癌化疗敏感性的理想指标.

Relationship between P-glycoprotein function in peripheral blood cells and multidrug resistance in breast carcinoma

Objective To analyze the relationship between P-glycoprotein function in peripheral blood ceils and primary multidrug resistance in breast carcinoma. Methods P-gp function was investigated by flow cytometry in NK cells of 16 breast cancer patients treated with anthracyclines and taxnes. Among all the patients, 8 were in chemotherapy-sensitive group and 8 in chemotherapy-resistant group. P-gp function was determined by rhodamine 123 (Rh123 ) -ejection test. Mathematical model was established by a regression of the fluorescence-time curve. The efflux rate constants of the chemotherapy-sensitive and -resistant groups were compared. Results There was no significant difference of Rh123 accumulation, retaintion or efflux between the two groups. The mathematical model of F1 = F0·e-kt was established. K was the efflux rate constant, which was significantly different between the chemotherapy-sensitive and -resistant groups (P =0.025). When k >3. 9 was used as diagnostic criterium for primary resistance, the sensitivity, specificity and accuracy were 75.0% , 100% and 87.5% , respectively. Conclusion P-glycoprotein function in peripheral blood cells is associated with primary multidrug resistance in breast carcinoma. The efflux rate constant may be a good predictor for chemotherapy sensitivity.