CAJ | 학술논문

目的研究慢性淋巴细胞白血病(CLL)患者脂蛋白酯酶(LPL)表达情况,探讨LPL在CLL预后中的意义.方法采用基于Taqman探针技术的实时定量逆转录PCR(qRT-PCR)方法检测62例CLL患者及10名健康对照者外周血标本中LPL的表达水平.对LPL表达水平与免疫球蛋白重链可变区(IgVH)基因突变、ZAP-70蛋白和CD38表达等预后因素进行相关性分析.采用ROC曲线确定LPL表达的最佳分界值,计算其对IgVH突变情况的阳性及阴性预测值.结果 qRT-PCR标准曲线的R2均≥0.990,敏感度可以检测到102拷贝/μg RNA,批内差及批间差变异系数(CV)均小于5%.62例CLL患者LPL表达水平中位数为0.006 0(0～0.737 0).在10名健康对照者中LPL均不表达或低表达,LPL中位表达水平为0(0～0.000 4).在经CD19磁珠分选后的3份CLL患者标本中,LPL相对表达量分别为0.036 0、0.075 0和0.197 0,与分选前表达水平(分别为0.024 0、0.074 0和0.225 0)相似.LPL表达水平与IgVH基因突变情况相关(r=0.45,P<0.05),无突变患者的LPL中位表达水平为0.006 0(0.000 7～0.110 0),明显高于突变患者的LPL中位表达水平[0.002 0(0.000 2～0.027 0)],差异有统计学意义(U=96.5,P<0.05);LPL表达水平与ZAP-70(r=0.38,P<0.05)及CD38(r=0.43,P<0.05)均显著相关.按照IgVH突变情况作为标准对LPL表达水平做ROC曲线分析,确定最佳分界值为0.036 0,敏感度为66.7%,特异度为72.4%,对IgVH突变情况阳性预测值(无突变)为51.8%,阴性预测值(有突变)为83.3%.结论 qRT-PCR方法检测LPL表达敏感可靠.LPL表达水平与CLL重要预后因素IgVH基因突变、ZAP-70、CD38密切相关,并可较准确地预测IgVH突变情况,在CLL的预后中具有重要价值.
목적 연구만성림파세포백혈병(CLL)환자지단백지매(LPL)표체정황,탐토LPL재CLL예후중적의의.방법 채용기우Taqman탐침기술적실시정량역전록PCR(qRT-PCR)방법검측62례CLL환자급10명건강대조자외주혈표본중LPL적표체수평.대LPL표체수평여면역구단백중련가변구(IgVH)기인돌변、ZAP-70단백화CD38표체등예후인소진행상관성분석.채용ROC곡선학정LPL표체적최가분계치,계산기대IgVH돌변정황적양성급음성예측치.결과 qRT-PCR표준곡선적R2균≥0.990,민감도가이검측도102고패/μg RNA,비내차급비간차변이계수(CV)균소우5%.62례CLL환자LPL표체수평중위수위0.006 0(0～0.737 0).재10명건강대조자중LPL균불표체혹저표체,LPL중위표체수평위0(0～0.000 4).재경CD19자주분선후적3빈CLL환자표본중,LPL상대표체량분별위0.036 0、0.075 0화0.197 0,여분선전표체수평(분별위0.024 0、0.074 0화0.225 0)상사.LPL표체수평여IgVH기인돌변정황상관(r=0.45,P<0.05),무돌변환자적LPL중위표체수평위0.006 0(0.000 7～0.110 0),명현고우돌변환자적LPL중위표체수평[0.002 0(0.000 2～0.027 0)],차이유통계학의의(U=96.5,P<0.05);LPL표체수평여ZAP-70(r=0.38,P<0.05)급CD38(r=0.43,P<0.05)균현저상관.안조IgVH돌변정황작위표준대LPL표체수평주ROC곡선분석,학정최가분계치위0.036 0,민감도위66.7%,특이도위72.4%,대IgVH돌변정황양성예측치(무돌변)위51.8%,음성예측치(유돌변)위83.3%.결론 qRT-PCR방법검측LPL표체민감가고.LPL표체수평여CLL중요예후인소IgVH기인돌변、ZAP-70、CD38밀절상관,병가교준학지예측IgVH돌변정황,재CLL적예후중구유중요개치.
Objective To investigate the expression level of lipoprotein lipase (LPL) mRNA in chronic lymphocytic leukemia (CLL) patients and evaluate the prognostic value of LPL in CLL Methods Quantitative real-time RT-PCR (qRT-PCR) was performed in 62 CLL patients, 10 normal controls using Taqman probe system. Association between LPL and other known prognostic factors, such as IgVH mutation status, ZAP-70 and CD38 expression, was determined using the Spearman correlation analysis. ROC curve was used to determine the cut-off value of LPL expression level, the positive and negative predictive value of IgVH mutation status. Results The correlation coefficients of the standard curves in qRT-PCR were not less than 0.990. The coefficients of variation (CV) of interrun assay and intramn assay were < 5%, and the sensitivity can reached 102 copies/μg RNA. The median LPL mRNA expression level was 0.006 0 (0-0.737 0) in 62 CLL patients, whereas in 10 normal controls LPL mRNA expression level was extremely low with the median level of 0 (0-0.000 4). The expression levels of LPL in three CLL samples after miniMACS-sorted CD19 positive B cells were 0.036 0, 0.075 0 and 0.197 0, which were similar to the levels before miniMACS-sorted (0.024 0, 0.074 0 and 0.225 0). LFL expression was significantly associated with IgVH mutation status (r=0.45, P<0.05) . LPL expression level in IgVH unmutated patients [0.006 0 (0.000 7-0.110 0)] was significantly higher than the level in IgVH mutated patients [0.002 0(0.000 2-0.027 0)] (U=96.5, P<0.05). LPL expression was also significantly associated with ZAP-70 (r=0.38, P<0.05), CD38 expressions (r=0.43, P<0.05). According to ROC curve, the cut-off of LPL mRNA expression level was 0.036, with a 66.7% specificity, a 72.4% sensitivity, a 51.8% positive predictive value (IgVH unmutated), and a 83.3% negative predictive value (IgVH mutated) for IgVH mutation status. Conclusions The qRT-PCR assay is reliable and sensitive. LPL mRNA expression significantly correlates with IgVH mutation status, ZAP-70 and CD38 expression, and could be a predictive marker of IgVH mutation status. Our data confirms a role for LPL as a novel prognostic indicator in CLL.