本文選題：STAT5B + T-ALL�。� 參考：《蘇州大學(xué)》2015年碩士論文

【摘要】：目的1、分析闡述蘇州大學(xué)附屬第一醫(yī)院急性T淋巴細(xì)胞白血病(T-ALL)的臨床特點(diǎn)及實(shí)驗(yàn)室特征。2、探討T-ALL患者及本中心保存的8種T-ALL細(xì)胞株STAT5B基因突變。3、篩選針對(duì)STAT5B基因突變的靶向藥物,初步闡述其作用機(jī)理。研究對(duì)象和方法1、研究對(duì)象:收集2006年1月至2014年7月在蘇州大學(xué)附屬第一醫(yī)院初診急性T淋巴細(xì)胞白血病(T-ALL)的141例患者的臨床及實(shí)驗(yàn)室資料。其中,男性116例,女性25例,男女性別比例為4.64:1;成年患者(18歲)93例,兒童患者("f18歲)48例,患者中位年齡為24歲。2、選取93例急性T淋巴細(xì)胞白血病患者及本中心保存的8種T-ALL細(xì)胞株,抽提RNA,并設(shè)計(jì)引物通過RT-PCR擴(kuò)增STAT5B基因全長,所得RT-PCR產(chǎn)物采用第一代測序技術(shù)Sanger測序法分析STAT5B基因突變。3、以伴有STAT5B N642H位點(diǎn)基因突變的KOPT-K1細(xì)胞株為細(xì)胞模型,應(yīng)用CCK8法分析初步篩選STAT5B N642H位點(diǎn)基因突變靶向藥物。4、應(yīng)用流式細(xì)胞術(shù)(flow cytometry,FCM)檢測候選藥物第三代TKI Ponatinib處理KOPT-K1細(xì)胞后的凋亡及周期變化。結(jié)果急性T淋巴細(xì)胞白血病患者臨床及實(shí)驗(yàn)室特征1、141例急性T淋巴細(xì)胞白血病患者,白細(xì)胞計(jì)數(shù)中位值為93×109/L,血紅蛋白含量中位值為105g/L,血小板計(jì)數(shù)中位值為60×109/L,骨髓原始細(xì)胞比例中位值為83.5%。按照FAB分型標(biāo)準(zhǔn),141例T-ALL患者中ALL-L1型40例,ALL-L2型46例,ALL未分類型(NOS)55例。三組患者臨床基本信息統(tǒng)計(jì)分析發(fā)現(xiàn),在初診骨髓原幼細(xì)胞比例中位值方面,ALL-L1ALL-L2NOS(88%84.5%76%,P0.05);在年齡、性別、中位白細(xì)胞計(jì)數(shù)、中位血紅蛋白含量、中位血小板計(jì)數(shù)方面,三組之間無統(tǒng)計(jì)學(xué)差異。2、141例T-ALL患者中,98.6%(139/141)的患者染色體核型分析成功,染色體正常核型(NK)占58.3%(81/139),異常核型檢出率為41.7%(58/139)。其中,單體核型(MK)占5.8%(8/139),t(11;14)(p13;q11)占5.0%(7/139),其它≥2次的結(jié)構(gòu)性異常主要有6q-,del(6)(q21),i(7q),+8。3、83例T-ALL患者進(jìn)行融合基因檢測,20例患者可檢測到基因異常。4例男性患者檢測到SET-NUP214融合基因轉(zhuǎn)錄本,中位年齡為26(23-54)歲;8例患者檢測到SIL-TAL1融合基因轉(zhuǎn)錄本,中位年齡為17.5(12-42)歲,男女性別比為7:1。其他檢測到的融合基因有:SET-CAN(+)2例,BCR-ABL(+)1例,HOX11(+)1例,MLL-MLL(+)1例,MLL-AF10(+)1例,MLL-AF9(+)1例,TCR重排(+)1例。4、對(duì)117例T-ALL患者進(jìn)行常見基因突變檢測(PHF6、FBXW7、NOTCH1、WT1、LMO2),其中,NOTCH1突變最常見,發(fā)生率為47%(55/117),16例患者具有PHF6突變(13.7%),FBXW7、WT1、LMO2突變例數(shù)分別為12例(10.2%)、4例(3.4%)、3例(2.6%)。急性T淋巴細(xì)胞白血病患者及T-ALL細(xì)胞株中STAT5B基因突變研究Sanger測序結(jié)果顯示93例T-ALL患者及8株T-ALL細(xì)胞株中只發(fā)現(xiàn)STAT5B N642H位點(diǎn)基因突變。其中,1例兒童患者伴有STAT5B p.N642H突變,8株T-ALL細(xì)胞株中KOPT-K1細(xì)胞株存在STAT5B N642H突變。STAT5B基因突變靶向藥物篩選1、4種候選藥物分析發(fā)現(xiàn),酪氨酸激酶抑制劑(TKI)及STAT5B抑制劑AZD1480對(duì)KOPT-K1細(xì)胞均有抑制作用。其中,第3代TKI Ponatinib對(duì)其抑制作用最強(qiáng),其抑制作用優(yōu)于STAT5B抑制劑AZD1480。2、第3代TKI Ponatinib處理KOPT-K1細(xì)胞72h后,流式細(xì)胞周期檢測發(fā)現(xiàn)G0/G1細(xì)胞比例明顯高于未加藥組(75.61±0.12%VS 58.88±4.07%,P0.05)。結(jié)論1、本中心141例急性T淋巴細(xì)胞白血病患者中,異常染色體及再現(xiàn)性染色體異常檢出率較低(41.7%),最常見的融合基因?yàn)镾IL-TAL1,NOTCH1突變在T-ALL中發(fā)生率最高。2、成人T-ALL患者中STAT5B N642H位點(diǎn)突變罕見,兒童中STAT5B N642H突變發(fā)生率為3.6%,發(fā)現(xiàn)KOPT-K1細(xì)胞系存在STAT5B N642H位點(diǎn)基因突變,為研究STAT5B基因突變的藥物提供理想的細(xì)胞模型。3、第三代TKI Ponatinib對(duì)KOPT-K1細(xì)胞的增殖有抑制作用,且KOPT-K1細(xì)胞對(duì)Ponatinib最為敏感,提示小分子藥物Ponatinib在治療STAT5B基因突變的血液病中可能具有臨床應(yīng)用價(jià)值。
[Abstract]:Objective 1 to analyze the clinical characteristics and laboratory characteristics of acute T lymphocytic leukemia (T-ALL) in First Hospital Affiliated to Suzhou University,.2, to explore the STAT5B gene mutation.3 of T-ALL patients and the 8 T-ALL cell lines preserved in this center, and to screen the target drugs for the mutation of STAT5B gene. The research object and method 1, the research object and method, are discussed. Objective: to collect the clinical and laboratory data of 141 patients with acute T lymphocytic leukemia (T-ALL) in First Hospital Affiliated to Suzhou University from January 2006 to July 2014, including 116 male and 25 female, 25 male and female sex ratio 4.64:1, 93 adult patients (18 years old), 48 cases of children ("F18 years old"), and the median age of 24 years.2, 93 patients with acute T lymphocytic leukemia and 8 T-ALL cell lines preserved in this center were selected to extract RNA, and primers were designed to amplify the full length of the STAT5B gene by RT-PCR. The RT-PCR products were analyzed by the first generation sequencing technique and Sanger sequencing method to analyze the STAT5B gene mutation.3, and the KOPT-K1 cell line with the STAT5B N642H site gene mutation was fine. Cell model, CCK8 method was used to preliminarily screen STAT5B N642H loci gene mutation targeting drug.4, and the apoptosis and cycle changes after third generation TKI Ponatinib processing KOPT-K1 cells were detected by flow cytometry (flow cytometry, FCM). Results of acute T lymphocyte leukopathy, 1141 cases of acute T lymphatic lymph nodes were characterized by clinical and laboratory characteristics. In leukemic patients, the median value of leukocyte count was 93 x 109/L, the median value of hemoglobin content was 105g/L, the value of the platelet count was 60 x 109/L, the median of bone marrow cell proportion was 83.5%. according to the FAB classification standard, 141 cases of T-ALL patients were ALL-L1 type, 46 cases of ALL-L2, 55 cases of ALL undivided (NOS). Information statistical analysis found that ALL-L1ALL-L2NOS (88%84.5%76%, P0.05) in the proportion of primary myeloid cells in the first diagnosis; there was no statistical difference between the three groups in age, sex, median white blood cell count, median hemoglobin content, and median platelet count, and 98.6% (139/141) of the patients with T-ALL. The chromosomal normal karyotype (NK) was 58.3% (81/139), and the abnormal karyotype detection rate was 41.7% (58/139). Among them, the monosomal karyotype (MK) accounted for 5.8% (8/139), t (11; 14) (P13; Q11) accounted for 5% (7/139), and the other 2 Structural abnormalities were mainly 6q-, del (6) (q21), and 20 patients could detect gene differentiation. The SET-NUP214 fusion gene transcriptional transcript was detected in.4 male patients, and the median age was 26 (23-54) years; 8 patients detected the SIL-TAL1 fusion gene transcriptional transcript, the median age was 17.5 (12-42) years, and the sex ratio of men and women was 7:1. (2 cases, +) 1 cases, HOX11 (+) 1 cases, MLL-MLL (+) 1 cases, MLL-AF10 (+) 1 cases, M, M, M, M, M, and M. LL-AF9 (+) 1 cases, TCR rearrangement (+) 1 cases of.4, 117 cases of T-ALL patients with common gene mutation detection (PHF6, FBXW7, NOTCH1, WT1, LMO2), of which NOTCH1 mutation is the most common, the incidence is 47% (55/117), 16 cases with PHF6 mutations (10.2%), 4 cases (3.4%), 3 cases (2.6%). Acute lymphoblastic leukemia. The Sanger sequencing of STAT5B gene mutations in patients and T-ALL cells showed that only 93 cases of T-ALL and 8 T-ALL cell lines found only the mutation of STAT5B N642H loci, of which 1 children were accompanied by STAT5B p.N642H mutation, and the KOPT-K1 cell line of 8 T-ALL cell lines had a mutant targeting drug sieve. The selection of 1,4 candidate drug analysis showed that tyrosine kinase inhibitor (TKI) and STAT5B inhibitor AZD1480 had inhibitory effect on KOPT-K1 cells. Among them, the third generation TKI Ponatinib had the strongest inhibitory effect. The inhibitory effect was better than the STAT5B inhibitor AZD1480.2, and the third generation TKI Ponatinib treated KOPT-K1 cells. The proportion of /G1 cells was significantly higher than that in the untreated group (75.61 + 0.12%VS 58.88 + 4.07%, P0.05). Conclusion 1. Among 141 patients with acute T lymphocytic leukemia, abnormal chromosomes and reproducible chromosomal abnormalities were low (41.7%), the most common fusion gene was SIL-TAL1, NOTCH1 mutation was the highest in T-ALL and S in adult T-ALL patients. The mutation of TAT5B N642H site is rare, and the incidence of STAT5B N642H mutation in children is 3.6%. It is found that there is a STAT5B N642H locus mutation in the KOPT-K1 cell line, which provides an ideal cell model.3 for the study of the STAT5B gene mutation. The third generation TKI Ponatinib can inhibit the proliferation of the KOPT-K1 cells, and the cells are most sensitive to the STAT5B. It is suggested that small molecule Ponatinib may be of clinical value in the treatment of STAT5B gene mutation hematopathy.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R733.71

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 崔森;母子同患急性B淋巴細(xì)胞白血病[J];中華血液學(xué)雜志;2001年01期

2 馬麗輝,楊波,喬振華,蘇麗萍,李建蘭;急性B淋巴細(xì)胞白血病伴嗜酸粒細(xì)胞增多癥一例[J];中華血液學(xué)雜志;2001年09期

3 周曉宏,童春容;自體細(xì)胞因子誘導(dǎo)的殺傷細(xì)胞治療急性B淋巴細(xì)胞白血病22例臨床觀察[J];河北醫(yī)學(xué);2003年04期

4 張伯偉,趙磊,郭如華,李秀珍,別立潔,楊瑞云;鄭州地區(qū)T淋巴細(xì)胞白血病病毒-Ⅰ感染情況調(diào)查[J];河南醫(yī)學(xué)研究;2003年02期

5 孔小行,柴憶歡,李建琴;40例兒童T淋巴細(xì)胞白血病MICM分型及其治療與預(yù)后分析[J];中國血液流變學(xué)雜志;2003年03期

6 陳正林;朱建一;余靜;;急性雙表型淋巴細(xì)胞白血病誤診為淋巴肉瘤細(xì)胞白血病1例[J];臨床檢驗(yàn)雜志;2006年02期

7 邢江濤;李剛榮;史冬霞;高爽;李慧慧;;花形核急性B淋巴細(xì)胞白血病一例[J];白血病.淋巴瘤;2007年06期

8 薛麗;汪盛;魯昌立;王婷婷;王琳;郭在培;;伴有皮膚表現(xiàn)的慢性B淋巴細(xì)胞白血病[J];臨床皮膚科雜志;2008年05期

9 陳宏安;門自起;;慢性B淋巴細(xì)胞白血病漏診一例[J];中國療養(yǎng)醫(yī)學(xué);2011年06期

10 曹盛;唐斌;吳昌松;朱廷倫;盧征翠;;貴州黔南地區(qū)人群T淋巴細(xì)胞白血病病毒Ⅰ/Ⅱ感染情況調(diào)查分析[J];檢驗(yàn)醫(yī)學(xué)與臨床;2012年13期

相關(guān)會(huì)議論文 前10條

1 王偉東;桑素風(fēng);李建華;;青少年患急性型T淋巴細(xì)胞白血病一例[A];中華醫(yī)學(xué)會(huì)第八次全國血液學(xué)學(xué)術(shù)會(huì)議論文匯編[C];2004年

2 ;慢性B淋巴細(xì)胞白血病共刺激分子表達(dá)的研究[A];2008年浙江省檢驗(yàn)醫(yī)學(xué)學(xué)術(shù)年會(huì)論文匯編[C];2008年

3 仇海榮;楊慧;王蓉;吳雨潔;李建勇;;熒光原位雜交技術(shù)在成人急性B淋巴細(xì)胞白血病中的應(yīng)用[A];第13屆全國實(shí)驗(yàn)血液學(xué)會(huì)議論文摘要[C];2011年

4 劉英;朱平;胡亞美;;急性B淋巴細(xì)胞白血病免疫球蛋白重鏈可變區(qū)基因的分子特征[A];第九屆全國實(shí)驗(yàn)血液學(xué)會(huì)議論文摘要匯編[C];2003年

5 劉英;朱平;胡亞美;;急性B淋巴細(xì)胞白血病免疫球蛋白重鏈可變區(qū)基因的分子特征[A];第三屆全國血液免疫學(xué)學(xué)術(shù)大會(huì)論文集[C];2003年

6 李可嘉;施若非;鄭捷;;伴有皮膚表現(xiàn)的慢性B淋巴細(xì)胞白血病[A];中華醫(yī)學(xué)會(huì)第十八次全國皮膚性病學(xué)術(shù)年會(huì)論文匯編[C];2012年

7 沈樹紅;蔣黎明;湯靜燕;薛惠良;陳靜;董璐;潘慈;胡文婷;劉秋霞;劉茵;羅成娟;顧龍君;;早期T前體細(xì)胞樣表型是兒童急性T淋巴細(xì)胞白血病的重要不良預(yù)后指標(biāo)[A];第六屆江浙滬兒科學(xué)術(shù)會(huì)議暨兒科學(xué)基礎(chǔ)與臨床研究進(jìn)展學(xué)術(shù)班論文匯編[C];2009年

8 楊艷萍;;原代急性T淋巴細(xì)胞白血病的生物學(xué)特性研究[A];第13屆全國實(shí)驗(yàn)血液學(xué)會(huì)議論文摘要[C];2011年

9 孫慧;王倩;李元堂;張之芬;武煥玲;田文君;劉斌;;145例成人急性B淋巴細(xì)胞白血病免疫表型分析[A];中華醫(yī)學(xué)會(huì)第七次全國中青年檢驗(yàn)醫(yī)學(xué)學(xué)術(shù)會(huì)議論文匯編[C];2012年

10 李可嘉;施若非;鄭捷;;伴有皮膚表現(xiàn)的慢性B淋巴細(xì)胞白血病[A];2012全國中西醫(yī)結(jié)合皮膚性病學(xué)術(shù)會(huì)議論文匯編[C];2012年

相關(guān)重要報(bào)紙文章 前1條

1 蔣明　戴勁松;Ｌｉｖｉｎ蛋白導(dǎo)致Ｔ淋巴細(xì)胞白血病抗藥[N];醫(yī)藥經(jīng)濟(jì)報(bào);2004年

相關(guān)博士學(xué)位論文 前2條

1 沈燕;肝再生增強(qiáng)因子在急性T淋巴細(xì)胞白血病中的表達(dá)及其功能的實(shí)驗(yàn)研究[D];重慶醫(yī)科大學(xué);2014年

2 康園園;染色質(zhì)相互作用介導(dǎo)的癌基因TAL1的特異性激活對(duì)急性T淋巴細(xì)胞白血病的調(diào)控機(jī)制[D];吉林大學(xué);2013年

相關(guān)碩士學(xué)位論文 前10條

1 馬曉霖;急性T淋巴細(xì)胞白血病中STAT5B基因突變及靶向藥物篩選[D];蘇州大學(xué);2015年

2 于永娟;淋巴細(xì)胞白血病相關(guān)差異基因表達(dá)譜整合分析[D];蘇州大學(xué);2010年

3 盧娟;免疫表型在慢性B淋巴細(xì)胞白血病診斷、鑒別診斷及預(yù)后評(píng)價(jià)中的意義研究[D];安徽醫(yī)科大學(xué);2009年

4 沈和萍;兒童急性T淋巴細(xì)胞白血病微小殘留病的監(jiān)測及臨床價(jià)值[D];浙江大學(xué);2010年

5 倪萬茂;三氧化二砷誘導(dǎo)T淋巴細(xì)胞白血病細(xì)胞株細(xì)胞凋亡及其分子機(jī)制研究[D];浙江大學(xué);2004年

6 吳春晨;趨化因子及其受體在B淋巴細(xì)胞白血病發(fā)病機(jī)制中的作用[D];武漢大學(xué);2005年

7 楊晨;CD25與急性B淋巴細(xì)胞白血病的相關(guān)性及臨床意義探討[D];山西醫(yī)科大學(xué);2014年

8 趙慧芳;耐門冬酰胺酶的急性T淋巴細(xì)胞白血病細(xì)胞生物學(xué)特性研究[D];鄭州大學(xué);2010年

9 付偉;LIM蛋白FHL1C對(duì)急性T淋巴細(xì)胞白血病的作用和機(jī)制研究[D];第四軍醫(yī)大學(xué);2011年

10 南禎;MicroRNA-223在淋巴細(xì)胞白血病原代細(xì)胞中表達(dá)及作用機(jī)制的研究[D];天津醫(yī)科大學(xué);2013年

，

本文編號(hào)：2014940