非霍奇金淋巴瘤患者骨髓单个核细胞BCL-2/IgH、IgH基因重排的检测

BCL-2/IgH and IgH Gene Rearrangments in Bone Marrow Mononuclear Cells of Patients with Non-Hodgkin′s Lymphoma

本研究检测非霍奇金淋巴瘤(NHL)患者BCL-2/IgH基因主要断裂区重排、IgH基因重排,并探讨其对疾病的早期诊断、疗效评价等方面的意义。分别提取70例NHL(60例B-NHL、10例T-NHL)、7例淋巴结炎性肿大患者、20名正常人的骨髓单个核细胞DNA,通过PCR方法检测BCL-2/IgH、IgH基因重排,以凝胶电泳出现相应条带者为阳性,并与患者病理组织学检查进行比较,探讨此两种基因重排发生的相关因素,比较化疗后的动态变化。结果表明,①30例弥漫大B细胞淋巴瘤患者(DLBCL)中,10例骨髓BCL-2/IgH重排阳性(33.3%),与30例其它B-NHL(除外滤泡淋巴瘤FL及DLBCL)阳性率(6.7%)、20名正常人阳性率(5%)比较均有显著性差异(p均<0.05)。所有T-NHL及7例淋巴结炎性肿大患者BCL-2/IgH重排均为阴性;②对8例BCL-2/IgH基因重排阳性患者进行动态监测,经2个疗程R-CHOP治疗后BCL-2/IgH基因重排明显减少,PCR半定量结果均值由初治0.59降至0.16(p<0.05),6个疗程R-CHOP治疗后PCR半定量均值为0,BCL-2/IgH基因重排完全转阴;③BCL-2/IgH基因重排阳性患者中LDH水平升高占81.8%,在重排阴性患者中占28.6%,两组间有统计学差异(p<0.05)。然而,BCL-2/IgH基因重排与淋巴瘤分期、是否伴有全身症状、β2-MG水平、骨髓侵犯、肝脏脾脏侵犯均无显著相关性;④20例DLBCL(均为初治)骨髓单个核细胞DNA检测显示,9例IgH基因重排阳性(45%);30例其它B-NHL(均为初治或复发患者,DLBCL除外)中14例IgH基因重排阳性(46.7%),其两组间无统计学差异性(p>0.05),但对照组20名正常人、10例T细胞淋巴瘤患者及7例淋巴结炎性肿大患者均为阴性;⑤对7例IgH基因重排阳性患者进行动态监测表明,1个疗程的R-CHOP治疗就能显著减少IgH基因重排,PCR半定量结果均值由初治0.42降至0.13(p<0.05),2个疗程后PCR半定量均值为0,重排完全消除;⑥IgH基因重排阳性患者中LDH水平升高占90%,在重排阴性患者中占30%,两组间有统计学差异(p<0.05);IgH基因重排与淋巴瘤分期、是否伴有全身症状、β2-MG水平、骨髓侵犯、肝脏脾脏侵犯均无显著相关性。结论:BCL-2/IgH、IgH基因重排均可作为B-NHL早期诊断及评价疗效的特异性指标,这两种重排均与LDH水平相关;BCL-2/IgH基因重排对DLBCL特异性较高。

This study was purposed to investigate the BCL-2/IgH gene rearrangement in major break point region（MBR） and IgH gene rearrangements of patients with non-Hodgkin＇s lymphoma（NHL）,and explore their significance for improving early diagnosis and accurately evaluating chemotherapy effect.DNA for BCL-2/IgH and IgH gene assays was extracted from bone marrow mononuclear cells in 70 cases of lymphoma（60 cases of B-NHL and 10 cases of T-NHL）,7 cases of lymphonode inflammary and 20 healthy controls.The BCL-2/IgH,IgH gene rearrangments were assayed by polymerase chain reaction（PCR）,the assayed results were compared with results of pathological biopsy;the factors related with occurrence of these 2 kinds of gene rearrangement were analyzed and the dynamic changes of BCL-2/IgH and IgH gene rearrangements after chemotherapy were compared,the chemotherapy effect was evaluated.The results indicated that（1） BCL-2/IgH gene rearrangement in bone marrow mononuclear cells was observed in 10 cases out of 30 DLBCL cases（33.3%）,and was more frequent than that in 30 other B-NHL cases（6.7%）,10 T-NHL cases（0%）,7 lymphonodes inflammary cases（0%）and 20 healthy controls（5%）（p〈0.05）.（2） the quantity of rearranged BCL-2/IgH gene of 8 DLBCL cases reduced from 0.59 to 0.16（p〈0.05） after 2 courses of R-CHOP chemotherapy and completely disappeared after 6 courses of R-CHOP chemotherapy.（3） 81.8% patients with BCL-2/IgH gene rearrangement showed high serum LDH level,while it was observed in 28.6% patients without this gene rearrangement（p〈0.05）.Lymphoma staging,systemic symptoms,β2-MG level,bone marrow involvement,infiltration of liver and spleen were not significantly correlated with BCL-2/IgH gene rearrangement.（4） IgH gene rearrangment was found in 9 cases out of 20 DLBCL patients（all newly diagnosed patients）（45%）,IgH rearrangment was observed in 14 cases out of 30 other B-NHL（all newly diagnostd or relaped patients,except patients with DLBCL）（46.7%） and there was no statistical difference between these 2 groups,however IgH rearrangement all were not observed in 20 healthy persons,10 T-NHL cases and 7 lymphonodes inflammatory cases.（5） the quantity of rearranged IgH gene in 7 DLBCL cases was reduced from 0.42 to 0.13 after one course of R-CHOP chemotherapy（p〈0.05） and completely disappeared after 2 courses of R-CHOP chemotherapy.（6） 90% patients with IgH gene rearrangement had high serum LDH level,while it was found in 30% patients without this gene rearrangement（p〈0.05）.Lymphoma staging,systemic symptoms,β2-MG levels,bone marrow involvement,infiltrationrs liver and spleen all were not significantly correlated with IgH gene rearrangement.It is concluded that the BCL-2/IgH and IgH gene rearrangements may be used as specific indicators in early diagnosis and accurate evaluation of therapy efficacy in B-NHL,these 2 kind of rearrangement correlate with LDH level.The BCL-2/IgH gene rearrangement is more specific for in DLBCL.