本文選題：CRLF2 + 突變��； 參考：《南京醫(yī)科大學(xué)》2017年碩士論文

【摘要】：第一部分成人急性淋巴細(xì)胞白血病CRLF2突變特點(diǎn)、表達(dá)水平及其臨床意義目的:細(xì)胞因子受體樣因子2(cytokine receptor-like factor 2,CRLF2)可促進(jìn)早期B淋巴細(xì)胞的增殖,對(duì)正常B淋巴細(xì)胞的發(fā)育起著重要的作用。本研究將探討成人急性淋巴細(xì)胞白血病(acute lymphoblastic leukemia,ALL)中CRLF2突變特點(diǎn)、表達(dá)水平及其與預(yù)后關(guān)系。方法:本研究通過(guò)聚合酶鏈?zhǔn)椒磻?yīng)(polymerasechainreaction,PCR)方法,特異性擴(kuò)增129例成人ALL患者CRLF2外顯子(exon)1-6,對(duì)擴(kuò)增產(chǎn)物進(jìn)行瓊脂糖凝膠電泳鑒定、DNA純化、sanger測(cè)序,并將測(cè)序結(jié)果與基因庫(kù)比對(duì),分析突變類型、突變發(fā)生率及其臨床特點(diǎn)。通過(guò)實(shí)時(shí)熒光定量PCR(real-time PCR)方法,檢測(cè)CRLF2表達(dá)水平。分析CRLF2在B-ALL、T-ALL以及正常對(duì)照之間可能存在的表達(dá)差異。分析CRLF2表達(dá)水平與IKZF1缺失可能存在的關(guān)系及其臨床意義。統(tǒng)計(jì)學(xué)分析采用SPSS 20.0軟件,計(jì)量資料采用非參數(shù)Mann-Whitney檢驗(yàn),計(jì)數(shù)資料采用χ2檢驗(yàn)(或Fisher精確檢驗(yàn)),采用Kaplan-Meier檢驗(yàn)對(duì)ALL患者進(jìn)行生存分析及預(yù)后關(guān)系研究。P0.05為差異有統(tǒng)計(jì)學(xué)意義。結(jié)果:本組CRLF2共檢測(cè)到6種基因改變類型,其中P224L、R186S提示預(yù)后良好,L86I、F232F、W255C可能與預(yù)后不良有關(guān)。在無(wú)IKZF1缺失的患者中,A11A突變組的RFS明顯長(zhǎng)于野生型組。CRLF2在ALL患者表達(dá)水平明顯高于正常人。CRLF2高表患者預(yù)后較差,總生存期明顯低于CRLF2低表組。CRLF2表達(dá)水平與IKZF1缺失有關(guān),IKZF1缺失患者的CRLF2表達(dá)水平明顯高于無(wú)IKZF1缺失患者。提示CRLF2與IKZF1之間可能存在功能上的聯(lián)系。結(jié)論:CRLF2突變及表達(dá)水平異常在成人ALL的發(fā)生發(fā)展中起著重要作用,本組檢測(cè)到的新型突變R186S、P224L與預(yù)后良好有關(guān),CRLF2表達(dá)水平增高往往提示不良預(yù)后。第二部分 SH2B3在中國(guó)成人急性淋巴細(xì)胞白血病中的突變研究目的:SH2B3(Srchomology2B3)編碼SH2配體適應(yīng)蛋白LNK,可以抑制JAK-STAT信號(hào)通路和細(xì)胞因子信號(hào)通路的激活,SH2B3的失活突變,可以導(dǎo)致JAK-STAT信號(hào)通路激活,從而引起急性淋巴細(xì)胞白血病(acute lymphoblastic leukemia,ALL)的發(fā)生。本組將研究成人ALL患者SH2B3突變特征及其臨床意義。方法:使用PCR及巢式PCR的方法,特異性擴(kuò)增SH2B3外顯子,其后進(jìn)行DNA純化、測(cè)序,測(cè)序結(jié)果與NCBI-BLAST進(jìn)行比對(duì),分析突變類型、突變發(fā)生率及其臨床特點(diǎn)。統(tǒng)計(jì)學(xué)分析采用SPSS 20.0軟件,計(jì)量資料采用非參數(shù)Mann-Whitney檢驗(yàn),計(jì)數(shù)資料采用χ2檢驗(yàn)(或Fisher精確檢驗(yàn)),采用二元Logistic回歸、Kaplan-Meier檢驗(yàn)、COX回歸對(duì)ALL患者進(jìn)行生存分析及預(yù)后關(guān)系研究。P0.05認(rèn)為有統(tǒng)計(jì)學(xué)意義。結(jié)果:本組107例初發(fā)ALL患者共檢測(cè)到兩種基因改變類型,c.724CT,p.P242S及c.784TC,p.W262R。經(jīng)過(guò)比對(duì),兩種基因改變均為單核苷酸多態(tài)性(Single nucleotide polymorphisms,SNP)類型,分別為 rs3184504 和 rs78894077。其中SH2B3P242S基因改變組的EFS、OS與野生型組相比,無(wú)統(tǒng)計(jì)學(xué)意義的延長(zhǎng)。在無(wú)IKZF1缺失的患者中,P242S基因改變組的EFS、OS長(zhǎng)于野生型組,差異有統(tǒng)計(jì)學(xué)意義,提示P242S是一種預(yù)后良好的基因改變類型。而W262R基因改變患者多次復(fù)發(fā),包括髓內(nèi)復(fù)發(fā)及中樞復(fù)發(fā),預(yù)后較差,因此認(rèn)為W262R與復(fù)發(fā)以及預(yù)后不良相關(guān)。結(jié)論:SH2B3突變?cè)诔扇薃LL的發(fā)生發(fā)展中起著重要作用,本組檢測(cè)到的兩種SNP類型可能與患者的預(yù)后相關(guān),有望成為新的分子預(yù)后靶點(diǎn)。
[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.
【學(xué)位授予單位】：南京醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R733.71

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 肖麗嬋;李敏;葛崢;顧巖;周喜連;郭星;李建勇;;IL-7R在成人急性淋巴細(xì)胞白血病中的突變及其臨床意義[J];中國(guó)實(shí)驗(yàn)血液學(xué)雜志;2016年04期

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本文編號(hào)：1936478