發(fā)布時(shí)間：2018-03-06 17:54

  本文選題：多發(fā)性骨髓瘤　切入點(diǎn)：DJ-1　出處：《川北醫(yī)學(xué)院》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:探討DJ-1基因在初診的多發(fā)性骨髓瘤(MM)患者CD138+瘤細(xì)胞中的表達(dá)情況,并分析DJ-1表達(dá)水平與臨床的相關(guān)性。方法:收集初診MM患者的骨髓液及正常人外周血,通過密度梯度離心法分離骨髓單個(gè)核細(xì)胞(BM-MNCs)和正常人外周血單個(gè)核細(xì)胞(PB-MNCs)。采用磁珠分選技術(shù),分離BM-MNCs中的CD138+瘤細(xì)胞。采用逆轉(zhuǎn)錄熒光定量PCR(RTq-PCR)檢測CD138+瘤細(xì)胞及正常人PB-MNCs的DJ-1 mRNA表達(dá)水平;采用蛋白免疫印跡法(Western blot)檢測相應(yīng)DJ-1蛋白的表達(dá)情況。收集患者的臨床信息,分析CD138+瘤細(xì)胞中DJ-1 mRNA表達(dá)水平與患者分型、初診時(shí)的國際評(píng)分系統(tǒng)(ISS)分期、乳酸脫氫酶(LDH)水平的關(guān)系。對(duì)患者的治療情況進(jìn)行隨訪并評(píng)價(jià)療效,分析基線水平的DJ-1 mRNA表達(dá)水平與療效是否相關(guān)。結(jié)果:1、共收集初診MM骨髓標(biāo)本28例,其中成功分選出可供分析的CD138+瘤細(xì)胞的有23例。2、MM患者CD138+瘤細(xì)胞DJ-1 mRNA相對(duì)表達(dá)水平=0.197(0.117-0.408)(2-ΔCT,n=23),正常人PB-MNCs DJ-1 mRNA相對(duì)表達(dá)水平為0.019(0.017-0.021)(2-ΔCT,n=23),差異具有顯著統(tǒng)計(jì)學(xué)意義(P0.001),并經(jīng)DJ-1蛋白檢測驗(yàn)證;3、男性和女性MM患者CD138+細(xì)胞內(nèi)DJ-1 mRNA相對(duì)表達(dá)水平分別為0.232(0.095-0.511)(2-ΔCT,n=16),0.197(0.133-0.307)(2-ΔCT,n=7),差異無統(tǒng)計(jì)學(xué)意義(P=0.764);"g65歲和65歲MM患者CD138+細(xì)胞內(nèi)dj-1mrna相對(duì)表達(dá)水平分別為0.285(0.164-0.408)(2-Δct,n=10),0.168(0.112-0.307)(2-Δct,n=13),差異無統(tǒng)計(jì)學(xué)意義(p=0.336);4、igg型和iga型mm患者cd138+細(xì)胞內(nèi)dj-1mrna相對(duì)表達(dá)水平分別為0.178(0.133-0.510)(2-Δct,n=10),0.273(0.095-0.408)(2-Δct,n=9),差異無統(tǒng)計(jì)學(xué)意義(p=0.806)。同樣,輕鏈κ型與λ型的mm患者cd138+細(xì)胞內(nèi)dj-1mrna相對(duì)表達(dá)水平分別為0.249(0.112-0.558)(2-Δct,n=12),0.192(0.117-0.307)(2-Δct,n=11),差異無統(tǒng)計(jì)學(xué)意義(p=0.460)5、mm患者cd138+細(xì)胞內(nèi)dj-1mrna相對(duì)表達(dá)水平在issi+ii期和iii期分別為0.273(0.164-0.343)(2-Δct,n=11),0.192(0.117-0.307)(2-Δct,n=11);6、初診時(shí)issi+ii和iii期患者ldh(u/l)水平分別為138.1(124.8-197)(u/l,n=10),171.8(126-214.5)(u/l,n=7),dj-1mrna相對(duì)表達(dá)量(2-Δct)與ldh相關(guān)系數(shù)為0.458(n=17);7、化療2周期后療效達(dá)部分緩解及以上(≥pr)的患者和未達(dá)pr的患者,其基線dj-1mrna相對(duì)表達(dá)水平分別為0.51(0.117-0.558)(2-Δct,n=5),0.273(0.112-0.307)(2-Δct,n=7)。結(jié)論:1、mm患者cd138+瘤細(xì)胞dj-1表達(dá)水平異常增高;2、mm患者cd138+瘤細(xì)胞dj-1mrna表達(dá)水平與患者的性別、是否大于65歲、疾病分型不相關(guān);3、由于病例數(shù)少,隨訪觀察時(shí)間短,mm患者cd138+瘤細(xì)胞dj-1mrna表達(dá)水平與與初診時(shí)患者的疾病分期、ldh水平、療效的關(guān)系尚不確定,需要進(jìn)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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