Objective To explore the feasibility and significance of detecting Cyclin D1 and bcl-2 with flow cytometry in the diagnosis of B-cell lymphoma.Methods 45 patients patients with confirmed hematologic diseases in final diagnosis were collected and recorded 45 cases in detail included in the study.25 healthy persons showing normal results in routine physical examinations and laboratory tests were also selected and matched according to the age and gender.In addition,according to the age and gender matching principle,selected routine physical examination and all tests were within the normal range in 25 healthy persons as a healthy control group.Select the pathologically confirmed Cyclin D1 or bcl-2 positive cases were selected as the positive control group,select and the negative cases as the negative control group.Four-color monoclonal antibodies directly immunofluorescence monoclonal antibody labelinged with immunofluorescence were used to analyze the surface and cytoplasmic antigens,and multi-parameter flow cytometry was used to investigate the peripheral blood Cyclin D1 and bcl-2 subsets in lymphoma patients.Results The normal values of Cyclin D1 MFI and bcl-2 were obtained from the 25 normal volunteers by analyzing the (x)±s values and 95 ％ confidence interval,thus establishing the diagnostic criteria.Compared to the healthy control (0.356±0.159,1.938±0.324),all B-cell lymphoma patients had increased expression in Cyclin D1 (1.824±0.312)and bcl-2 MFI (4.259±0.541) in their peripheral blood,results showing statistical significance (P ＜ 0.01).Patients with different subtypes of lymphoma expressed differing Cyclin D1 MFI.In patients with Hodgkin' s lymphoma,Cyclin D1 MFI (0.386±0.112) was significantly lower than that in the non-Hodgkin' s lymphoma patients (1.623±1.987) (P ＜ 0.01).Further,in non-Hodgkin' s lymphoma patients,mantle cell lymphoma was 100 ％ positive for Cyclin D1,while the remaining subtypes were negative.Patients with different subtypes of lymphoma also showed differences in bcl-2 expression.In the Hodgkin' s lymphoma,bcl-2 (2.045±0.877) was significantly lower than the non-Hodgkin' s lymphoma (4.045±0.499) (P ＜ 0.01).In non-Hodgkin' s lymphoma,T cell lymphoma expressed the lowest,while MCL and FL showed 100 ％ positivity.In patients with positive Cyclin D1 or bcl-2,the mean fluorescence intensity level of Cyclin D1 (before treatment 3.099±0.349; after treatment 1.008±0.279) or bcl-2 (before treatment 7.814± 1.030,after treatment 3.131±0.522) was significantly lowered after treatment (P ＜ 0.01).Conclusion Using the method of flow cytometry for detecting Cyclin D1 and bcl-2 in lymphoma cells is feasible,and it can be applied clinically to evaluate the treatment.%目的 研究流式细胞术(FCM)检测Cyclin D1和bcl-2在B细胞淋巴瘤诊断中的应用、可行性及临床意义.方法 以收治确诊的45例淋巴瘤患者为研究对象.按照年龄、性别相匹配的原则,以各项检验指标均在正常范围内的25名健康体检者作为健康对照组.同时选取病理科已确诊的Cyclin D1或bcl-2阳性病例为阳性对照组,阴性病例为阴性对照组.采用四色直接免疫荧光素法标记细胞抗原,多参数FCM检测各亚型淋巴瘤患者外周血、骨髓或淋巴结Cyclin D1及bcl-2表达水平.结果 对健康对照组Cyclin D1 MFI及bcl-2 MFI进行分析后得出(x)±s值及95％可信区间,从而得出FCM检测两者的正常参考值范围.B细胞淋巴瘤患者Cyclin D1和bcl-2的表达水平分别为1.824±0.312、4.259±0.541,较健康对照组(0.356±0.159,1.938±0.324)明显升高,两者差异均有统计学意义(P＜0.01).霍奇金淋巴瘤(HL)患者Cyclin D1 MFI(0.386±0.112)明显低于非霍奇金淋巴瘤(NHL)患者(1.623±1.987)(P＜0.01),而在非霍奇金淋巴瘤(NHL)患者中,套细胞淋巴瘤(MCL)患者Cyclin D1阳性率最高(100％),其余亚型均为阴性；HL患者bcl-2表达水平(2.045±0.877)明显低于NHL患者(4.045±0.499)(P＜ 0.01),在NHL中,T细胞淋巴瘤均为阴性,滤泡性淋巴瘤、MCL患者bcl-2表达阳性率最高(100％).在Cyclin D1或bcl-2阳性的淋巴瘤患者中,发现治疗后Cyclin D1(治疗前3.099±0.349,治疗后1.008 ±0.279)或bcl-2(治疗前7.814±1.030,治疗后3.131±0.522)的平均荧光强度均有显著降低(P＜0.01).结论 FCM检测淋巴瘤细胞中Cyclin D1及bcl-2表达的方法简便可行,其在不同淋巴瘤中表达不一,可以作为临床观察治疗效果的指标之一.
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