摘要
目的 定量分析B细胞非霍奇金淋巴瘤（B-NHL）各期患者化疗前或后外周血单个核胞和血浆中增殖诱导配体（APRIL）mRNA及蛋白的表达水平,探讨APRIL在B-NHL中的意义.方法 采用实时荧光定量聚合酶链反应（RFQ-PCR）及酶联免疫吸附试验（ELISA）方法,根据标准曲线计算待测样本中靶mRNA及蛋白的准确含量.结果 RFQ-PCR检测靶基因mRNA含量的线性范围为101～109 pg/ml,批内和批间重复性测定的变异系数（CV）分别为1.69%～5.99%和6.35%～10.12%.采用ELISA方法检测靶蛋白含量,标准曲线r=0.9922.化疗前后B-NHL各期患者APRILmRNA及蛋白表达均显著高于正常对照（P＜0.01）,而不同分期化疗后Ⅲ期及Ⅳ期患者表达低于化疗前（P＜0.05）,但Ⅰ和Ⅱ期化疗前后比较差异无统计学意义（P＞0.05）.结论 APRILmRNA与蛋白表达水平相一致,其参与B-NHL的发生与发展.APRIL可能与肿瘤负荷量有关,且可能成为有效治疗B-NHL的新靶点.
Objective To investigate mRNA and protein expression of aproliferation-inducing ligand （APRIL） in peripheral blood mononuclear cell and plasma of patients with B cells non-Hodgkin lymphoma （B-NHL） pre- or post- chemotherapy and to explore the role of APRIL in the B-NHL. Methods The mRNA and protein expression of APRIL were detected by real-time fluorescence quantitative polymerase chain reaction （RFQ-PCR） and enzyme-linked immunosorbent assay（ELISA）, respectively. According to the standard curves,the quantitative levels of target mRNA and protein were determined. Results The detection linear range of targeted mRNA by RFQ-PCR was 101-109 pg/ml, and the coefficient of variation values for both intra- and inter-experimental reproducibility ranged 1.69 %-5.99 % and 6.35 %-10.12 %, respectively. To detect expression of APRIL protein by ELISA, the correlation coefficient of standard curves reached 0.9922. Before or after chemical treatment, the expression levels of APRIL mRNA and protein in patients with B-NHL were significantly higher than those in normal control （P 〈0.01）, while the expression levels in post-treatment patients with Ⅲ and Ⅳ stage were lower than those of pre-treatment patients with corresponding stage （P 〈0.05, respectively）, but those of pre- and post-treatment patients with Ⅰ / Ⅱ stage were not different （P 〉0.05）.Conclusion The expression level of APRIL mRNA is similar to that of APRIL protein. APRIL may be involved in pathogenesis and development of B-NHL. Moreover, APRIL may be related to the burden of B-NHL and it may be considered as a targeted molecule for B-NHL.
出处
《白血病．淋巴瘤》
CAS
2010年第8期479-482,共4页
Journal of Leukemia & Lymphoma
关键词
增殖诱导配体
B细胞淋巴瘤
实时荧光定量聚合酶链反应
酶联免疫吸附测定
Aproliferation-inducing ligand （APRIL）
Lymphoma B-cell
the real-time fluorescence quan-titative polymerase chain reaction （RFQ-PCR）
Enzyme-linked immunosorbe-nt assay（ELISA）
