手术前后乳腺癌患者外周血淋巴细胞端粒酶活性检测及临床意义

Peripheral Blood Lymphocytes Telomerase Activity in Breast Cancer Patients Before and After Radical Mastectomy

目的 检测乳腺癌根治术前后患者外周血淋巴细胞端粒酶活性的表达。方法 利用端粒重复序列扩增技术(TRAP)分别结合非变性聚丙烯酰胺凝胶电泳(Native -PAGE)和酶联免疫吸附试验(ELISA)两种方法检测乳腺癌患者外周血淋巴细胞端粒酶活性。36例端粒酶阳性乳腺癌患者,术前1～3天及术后8～1 0天各取外周静脉血标本一次,经端粒重复序列扩增技术(TRAP)扩增,所得样本分别用非变性聚丙烯酰胺凝胶电泳(Native -PAGE)定性检测和酶联免疫吸附试验(ELISA)定量检测端粒酶活性。ELISA法检测各样本的吸光度(ΔA) ,将同一时相样品的吸光度(ΔA)分为一组,用方差分析法进行组样品ΔA均数间的比较。结果 Native -PAGE定性检测结果:术前各样本凝胶电泳染色后有明显间隔6bp条带;术后各样本在电泳图上的条带较术前减弱。ELISA定量检测结果:样品的吸光度(ΔA) ,术前1～3天为0 .35 0±0 .0 2 6,术后8～1 0天为0 .2 5 1±0 .0 2 3,两者相比,差异有显著性(P <0 .0 0 1 )。结论 ( 1 )乳腺癌根治手术可以明显降低乳腺癌患者外周血淋巴细胞端粒酶的活性表达。( 2 )外周血端粒酶活性可能作为乳腺癌判断疗效的指标。

Objective To evaluate the clinical significance of peripheral blood lymphocytes telomerase activity in breast cancer before and after radical mastectomy. Methods Telomerase activity in peripheral blood lymphocytes of patients with breast cancer was detected through telomerase repeat amplification protocol (TRAP) combined with the non-quantitative detection of Native-PAGE silver staining and the quantitative detection of ELISA , among which 36 breast cancer patients with positive-telomerase were studied .Blood samples were obtained from all telomerase -positive patients in two successive times: preoperation, the 8th to 10th day after operation from which 2 groups for samples , namely preoperative group and postoperative group ,were formed. Detection of Native-PAGE silver staining and ELISA, after comparing the response relationship with analytical method ,were adopted here to detect the telomerase activity and to check and measure the net increase of absorbance for the sample(ΔA)respectively. Results (1)The results of Native-PAGE: In samples of preoperation, ladders of products with 6 bp increments were visible in PAGE silver staining; but they were significantly reduced in the samples of postoperational ones. (2)The results of ELISA: Significant difference between preoperative group and postoperative group (P<0.001) was apparent here with ΔA of 2 groups 0.350±0.026 and 0.251±0.023 respectively. Conclusion (1)The radical mastectomy can reduce the expression of telomerase activity of breast cancer patients evidently.(2)Detection of telomerase activity in peripheral blood lymphocytes in breast tumor patients might be a valuable parameter for assessing the effect of treatment. Its possible association with prognosis, follow-up and recurrence of breast cancer need to be further studied.