CAJ | 학술논문

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.
목적 정량분석B세포비곽기금림파류(B-NHL)각기환자화료전혹후외주혈단개핵포화혈장중증식유도배체(APRIL)mRNA급단백적표체수평,탐토APRIL재B-NHL중적의의.방법 채용실시형광정량취합매련반응(RFQ-PCR)급매련면역흡부시험(ELISA)방법,근거표준곡선계산대측양본중파mRNA급단백적준학함량.결과 RFQ-PCR검측파기인mRNA함량적선성범위위101～109 pg/ml,비내화비간중복성측정적변이계수(CV)분별위1.69%～5.99%화6.35%～10.12%.채용ELISA방법검측파단백함량,표준곡선r=0.9922.화료전후B-NHL각기환자APRILmRNA급단백표체균현저고우정상대조(P＜0.01),이불동분기화료후Ⅲ기급Ⅳ기환자표체저우화료전(P＜0.05),단Ⅰ화Ⅱ기화료전후비교차이무통계학의의(P＞0.05).결론 APRILmRNA여단백표체수평상일치,기삼여B-NHL적발생여발전.APRIL가능여종류부하량유관,차가능성위유효치료B-NHL적신파점.
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.