成人急性淋巴细胞白血病预后分子标记CRLF2和SH2B3特征研究

发布时间：2018-05-26 08:15

本文选题：CRLF2 + 突变　；参考：《南京医科大学》2017年硕士论文

【摘要】：第一部分成人急性淋巴细胞白血病CRLF2突变特点、表达水平及其临床意义目的:细胞因子受体样因子2(cytokine receptor-like factor 2,CRLF2)可促进早期B淋巴细胞的增殖,对正常B淋巴细胞的发育起着重要的作用。本研究将探讨成人急性淋巴细胞白血病(acute lymphoblastic leukemia,ALL)中CRLF2突变特点、表达水平及其与预后关系。方法:本研究通过聚合酶链式反应(polymerasechainreaction,PCR)方法,特异性扩增129例成人ALL患者CRLF2外显子(exon)1-6,对扩增产物进行琼脂糖凝胶电泳鉴定、DNA纯化、sanger测序,并将测序结果与基因库比对,分析突变类型、突变发生率及其临床特点。通过实时荧光定量PCR(real-time PCR)方法,检测CRLF2表达水平。分析CRLF2在B-ALL、T-ALL以及正常对照之间可能存在的表达差异。分析CRLF2表达水平与IKZF1缺失可能存在的关系及其临床意义。统计学分析采用SPSS 20.0软件,计量资料采用非参数Mann-Whitney检验,计数资料采用χ2检验(或Fisher精确检验),采用Kaplan-Meier检验对ALL患者进行生存分析及预后关系研究。P0.05为差异有统计学意义。结果:本组CRLF2共检测到6种基因改变类型,其中P224L、R186S提示预后良好,L86I、F232F、W255C可能与预后不良有关。在无IKZF1缺失的患者中,A11A突变组的RFS明显长于野生型组。CRLF2在ALL患者表达水平明显高于正常人。CRLF2高表患者预后较差,总生存期明显低于CRLF2低表组。CRLF2表达水平与IKZF1缺失有关,IKZF1缺失患者的CRLF2表达水平明显高于无IKZF1缺失患者。提示CRLF2与IKZF1之间可能存在功能上的联系。结论:CRLF2突变及表达水平异常在成人ALL的发生发展中起着重要作用,本组检测到的新型突变R186S、P224L与预后良好有关,CRLF2表达水平增高往往提示不良预后。第二部分 SH2B3在中国成人急性淋巴细胞白血病中的突变研究目的:SH2B3(Srchomology2B3)编码SH2配体适应蛋白LNK,可以抑制JAK-STAT信号通路和细胞因子信号通路的激活,SH2B3的失活突变,可以导致JAK-STAT信号通路激活,从而引起急性淋巴细胞白血病(acute lymphoblastic leukemia,ALL)的发生。本组将研究成人ALL患者SH2B3突变特征及其临床意义。方法:使用PCR及巢式PCR的方法,特异性扩增SH2B3外显子,其后进行DNA纯化、测序,测序结果与NCBI-BLAST进行比对,分析突变类型、突变发生率及其临床特点。统计学分析采用SPSS 20.0软件,计量资料采用非参数Mann-Whitney检验,计数资料采用χ2检验(或Fisher精确检验),采用二元Logistic回归、Kaplan-Meier检验、COX回归对ALL患者进行生存分析及预后关系研究。P0.05认为有统计学意义。结果:本组107例初发ALL患者共检测到两种基因改变类型,c.724CT,p.P242S及c.784TC,p.W262R。经过比对,两种基因改变均为单核苷酸多态性(Single nucleotide polymorphisms,SNP)类型,分别为 rs3184504 和 rs78894077。其中SH2B3P242S基因改变组的EFS、OS与野生型组相比,无统计学意义的延长。在无IKZF1缺失的患者中,P242S基因改变组的EFS、OS长于野生型组,差异有统计学意义,提示P242S是一种预后良好的基因改变类型。而W262R基因改变患者多次复发,包括髓内复发及中枢复发,预后较差,因此认为W262R与复发以及预后不良相关。结论:SH2B3突变在成人ALL的发生发展中起着重要作用,本组检测到的两种SNP类型可能与患者的预后相关,有望成为新的分子预后靶点。
[Abstract]:The first part of the CRLF2 mutation, expression level and its clinical significance in adult acute lymphoblastic leukemia: cytokine receptor like factor 2 (cytokine receptor-like factor 2, CRLF2) can promote the proliferation of early B lymphocytes and play an important role in the development of normal B lymphocytes. This study will discuss adult acute lymphoid lymphocytic lymphocytic lymphocytic development. The characteristics of CRLF2 mutation in acute lymphoblastic leukemia (ALL) and the relationship between the expression level and the prognosis. Methods: This study specifically amplified the CRLF2 exon (exon) 1-6 of 129 adult ALL patients by polymerase chain reaction (polymerasechainreaction, PCR). The amplified products were identified by agarose gel electrophoresis and DNA purification. Sanger sequencing, and comparison of the sequencing results with the gene pool, analyzed the mutation type, mutation rate and its clinical characteristics. The expression level of CRLF2 was detected by real-time fluorescence quantitative PCR (real-time PCR) method. The possible differences in the expression of CRLF2 between B-ALL, T-ALL and normal controls were analyzed. The expression level of CRLF2 and the absence of IKZF1 may be analyzed. The relationship and its clinical significance. Statistical analysis used SPSS 20 software, measurement data using non parametric Mann-Whitney test, counting data using chi 2 test (or Fisher accurate test), Kaplan-Meier test for the survival analysis of ALL patients and the relationship between the prognosis of.P0.05 is statistically significant. Results: this group of CRLF2 examination 6 types of gene change were detected, of which P224L, R186S suggested that the prognosis was good, L86I, F232F, W255C may be associated with poor prognosis. In the patients without IKZF1 deletion, the RFS of the A11A mutation group was significantly longer than that in the wild type group, and the expression level of.CRLF2 in ALL patients was significantly higher than that in the normal human.CRLF2 high table, and the total survival time was significantly lower than that of the CRLF2 lower table. The expression level of.CRLF2 was associated with the deletion of IKZF1, and the level of CRLF2 expression in patients with IKZF1 deletion was significantly higher than that without IKZF1. It suggested that there might be a functional relationship between CRLF2 and IKZF1. Conclusion: CRLF2 mutation and abnormal expression level play an important role in the development and development of adult ALL, the new mutation R186S, P224L in this group The high expression level of CRLF2 often suggests bad prognosis. Second the mutation of SH2B3 in Chinese adult acute lymphoblastic leukemia: the SH2B3 (Srchomology2B3) encoded SH2 ligand adapted to the protein LNK, which can inhibit the activation of the JAK-STAT signaling pathway and the cell factor signaling pathway, and the inactivation mutation of SH2B3, It can cause JAK-STAT signaling pathway activation to cause acute lymphoblastic leukemia (acute lymphoblastic leukemia, ALL). This group will study the characteristics and clinical significance of SH2B3 mutation in adult ALL patients. Methods: using PCR and nested PCR methods, specific amplification of exons of SH2B3, followed by DNA purification, sequencing, sequencing, and sequencing results. Comparison with NCBI-BLAST, analysis of mutation type, mutation rate and its clinical characteristics. Statistical analysis used SPSS 20 software, measurement data using non parametric Mann-Whitney test, counting data using chi 2 test (or Fisher accurate test), two yuan Logistic regression, Kaplan-Meier test, COX regression for survival analysis of ALL patients The relationship between.P0.05 and prognosis was statistically significant. Results: two kinds of gene change types were detected in 107 patients with primary ALL, c.724CT, p.P242S and c.784TC, p.W262R. after comparison, and the two gene changes were single nucleotide polymorphisms (Single nucleotide polymorphisms, SNP), which were rs3184504 and rs78894077.. The EFS and OS in the SH2B3P242S gene change group had no statistically significant extension compared with the wild type group. In the patients with no IKZF1 deletion, the EFS in the P242S gene change group and the OS were longer than those in the wild type. The difference was statistically significant, suggesting that P242S is a kind of gene change type with good prognosis. And the W262R gene change patients relapsed, including intramedullary relapse. The recurrence and poor prognosis of the center are considered to be associated with W262R recurrence and poor prognosis. Conclusion: SH2B3 mutation plays an important role in the development of adult ALL. The two types of SNP detected in this group may be associated with the prognosis of the patients, and are expected to be a new molecular prognostic target.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R733.71

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