DJ-1基因在多发性骨髓瘤细胞的表达及临床意义

发布时间：2018-03-06 17:54

  本文选题：多发性骨髓瘤　切入点：DJ-1　出处：《川北医学院》2017年硕士论文　论文类型：学位论文

【摘要】：目的:探讨DJ-1基因在初诊的多发性骨髓瘤(MM)患者CD138+瘤细胞中的表达情况,并分析DJ-1表达水平与临床的相关性。方法:收集初诊MM患者的骨髓液及正常人外周血,通过密度梯度离心法分离骨髓单个核细胞(BM-MNCs)和正常人外周血单个核细胞(PB-MNCs)。采用磁珠分选技术,分离BM-MNCs中的CD138+瘤细胞。采用逆转录荧光定量PCR(RTq-PCR)检测CD138+瘤细胞及正常人PB-MNCs的DJ-1 mRNA表达水平;采用蛋白免疫印迹法(Western blot)检测相应DJ-1蛋白的表达情况。收集患者的临床信息,分析CD138+瘤细胞中DJ-1 mRNA表达水平与患者分型、初诊时的国际评分系统(ISS)分期、乳酸脱氢酶(LDH)水平的关系。对患者的治疗情况进行随访并评价疗效,分析基线水平的DJ-1 mRNA表达水平与疗效是否相关。结果:1、共收集初诊MM骨髓标本28例,其中成功分选出可供分析的CD138+瘤细胞的有23例。2、MM患者CD138+瘤细胞DJ-1 mRNA相对表达水平=0.197(0.117-0.408)(2-ΔCT,n=23),正常人PB-MNCs DJ-1 mRNA相对表达水平为0.019(0.017-0.021)(2-ΔCT,n=23),差异具有显著统计学意义(P0.001),并经DJ-1蛋白检测验证;3、男性和女性MM患者CD138+细胞内DJ-1 mRNA相对表达水平分别为0.232(0.095-0.511)(2-ΔCT,n=16),0.197(0.133-0.307)(2-ΔCT,n=7),差异无统计学意义(P=0.764);"g65岁和65岁MM患者CD138+细胞内dj-1mrna相对表达水平分别为0.285(0.164-0.408)(2-Δct,n=10),0.168(0.112-0.307)(2-Δct,n=13),差异无统计学意义(p=0.336);4、igg型和iga型mm患者cd138+细胞内dj-1mrna相对表达水平分别为0.178(0.133-0.510)(2-Δct,n=10),0.273(0.095-0.408)(2-Δct,n=9),差异无统计学意义(p=0.806)。同样,轻链κ型与λ型的mm患者cd138+细胞内dj-1mrna相对表达水平分别为0.249(0.112-0.558)(2-Δct,n=12),0.192(0.117-0.307)(2-Δct,n=11),差异无统计学意义(p=0.460)5、mm患者cd138+细胞内dj-1mrna相对表达水平在issi+ii期和iii期分别为0.273(0.164-0.343)(2-Δct,n=11),0.192(0.117-0.307)(2-Δct,n=11);6、初诊时issi+ii和iii期患者ldh(u/l)水平分别为138.1(124.8-197)(u/l,n=10),171.8(126-214.5)(u/l,n=7),dj-1mrna相对表达量(2-Δct)与ldh相关系数为0.458(n=17);7、化疗2周期后疗效达部分缓解及以上(≥pr)的患者和未达pr的患者,其基线dj-1mrna相对表达水平分别为0.51(0.117-0.558)(2-Δct,n=5),0.273(0.112-0.307)(2-Δct,n=7)。结论:1、mm患者cd138+瘤细胞dj-1表达水平异常增高;2、mm患者cd138+瘤细胞dj-1mrna表达水平与患者的性别、是否大于65岁、疾病分型不相关;3、由于病例数少,随访观察时间短,mm患者cd138+瘤细胞dj-1mrna表达水平与与初诊时患者的疾病分期、ldh水平、疗效的关系尚不确定,需要进一步研究。
[Abstract]:Objective: to investigate the expression of DJ-1 gene in CD138 tumor cells of newly diagnosed patients with multiple myeloma (MM), and to analyze the correlation between the expression of DJ-1 and clinical manifestations. Methods: the bone marrow fluid of newly diagnosed MM patients and the peripheral blood of normal controls were collected. Bone marrow mononuclear cells (BM-MNCs) and normal peripheral blood mononuclear cells (PB-MNCsC) were isolated by density gradient centrifugation. CD138 tumor cells were isolated from BM-MNCs. The expression of DJ-1 mRNA in CD138 tumor cells and normal PB-MNCs was detected by reverse transcriptase fluorescence quantitative analysis (RT-PCR), and the expression of corresponding DJ-1 protein was detected by Western blot. The clinical information of the patients was collected. To analyze the relationship between the expression level of DJ-1 mRNA in CD138 tumor cells and the type of patients, the international scoring system (ISS) stage at first visit, and the level of lactate dehydrogenase (LDH). The patients were followed up and the curative effect was evaluated. To analyze whether the expression of DJ-1 mRNA at baseline level is related to the curative effect. Results 28 newly diagnosed MM bone marrow specimens were collected. The relative expression level of DJ-1 mRNA in CD138 tumor cells was 0.197 ~ 0.117-0.408 and 0.019 ~ 0.017 ~ 0.021 ~ (-1) T ~ (nn2323) in normal controls, respectively. The difference was statistically significant (P 0.001) and DJ-1 protein was used to detect PB-MNCs DJ-1 mRNA. The relative expression levels of DJ-1 mRNA in the CD138 cells of male and female MM patients were 0.232P 0.095-0.511C ~ 2- 螖 CTN ~ (16) (0.1970.133-0.307) ~ (2-) 螖 CTN ~ (7) with no statistical significance, "the relative expression levels of dj-1mrna in CD138 cells of patients aged 65 and 65 were 0.285n0.164-0.4082- 螖 CTT10100.1680.1 12-0.3072- 螖 CTT ~ (n13)", and there was no significant difference between them (P = 0.3364igg) and iga (P = 0.3364igg) and iga (P < 0.05), respectively, and there was no significant difference in the expression of dj-1mrna in CD138 cells between male and female patients with MM at the age of 0.285- 0.4082- 螖 CTTN = 0.1680.112-0.3072- 螖 CTN ~ (3367G) and iga (P = 0.33671g), respectively. The relative expression levels of dj-1mrna in cd138 cells of mm patients were 0.178 ~ (0.133-0.510) ~ (2-) 螖 CTN ~ (10) ~ (-) 0.273N 0.095-0.408 ~ ~ (2) ~ (-) ~ (-) ~ (2) ~ (-) ~ (2) ~ (?) dj-1mrna in cd138 cells, respectively. There was no significant difference in the expression of dj-1mrna between the two groups, and there was no significant difference between the two groups. The relative expression levels of dj-1mrna in cd138 cells of patients with light chain 魏 type and 位 type were 0.249 ~ (0.1) 12-0.558 ~ (-) 2- 螖 CTN ~ (1212) 0.1922 0.117-0.307 ~ (2) ~ (-)-螖 CTN ~ (2 +)-螖 CTN ~ (2 +). There was no significant difference in the relative expression levels of dj-1mrna in cd138 cells of patients with light chain 魏 type and 位 type in issi II stage and iii stage, respectively. The relative expression levels of dj-1mrna in cd138 cells in issi II stage and iii stage were 0.273 ~ 0.164-0.343anl2 -ctnn11a 0.1922 0.117-0.307 ~ (2) -螖 ctnnnn ~ (11), respectively. The relative expression levels of dj-1mrna in cd138 cells in issi II stage and iii stage were 0.273 ~ 0.164-0.343n ~ (-1), respectively. The correlation coefficient between the relative expression of dj-1mrna and ldh was 0.458N / L 177. the patients with partial remission (鈮，

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