CAJ | 학술논문

目的探讨巯基嘌呤甲基转移酶(TPMT)活性和其遗传多态性在儿童急性淋巴细胞白血病(ALL)个体化治疗中的临床意义.方法运用高效液相色谱法(HPLC)对100例初诊ALL患儿、180名健康儿童进行TPMT活性检测.采用在线Primer3软件设计引物,对30例具有临床事件的患儿进行TPMT基因DNA测序.分析TPMT活性、遗传多态性与临床不良反应的相关性.TPMT活性以在单位时间(1 h)内单位质量的血红蛋白(Hb,g)催化生成的2-氨基-6-甲基巯基嘌呤(6-MTG)的量(nmol)表示.结果 100例初诊ALL患儿TPMT活性为(31.72±10.31)nmol·g-1Hb·h-1,180名健康儿童TPMT活性为(30.70±9.67)nmol·g-1Hb· h-1,两者TPMT活性差异无统计学意义(P=0.450).19例发生严重骨髓抑制的患儿TPMT活性为(20.96±7.24) nmol·g-1Hb·h-1,6例ALL复发者初诊时TPMT活性为(40.46±8.18) nmol·g-1 Hb·h-1,两者分别与30例具有临床事件的ALL患儿初诊时活性[(24.07±11.43)nmol·g-1Hb·h-1]相比差异具有统计学意义(P值均＜0.05).5例化疗后发生严重肝脏毒性者初诊时TPMT活性为(23.60±7.48) nmol·g-1Hb·h-1,与30例ALL患儿初诊时活性相比差异无统计学意义(P =0.930).对30例具有临床事件的ALL患儿TPMT基因DNA测序,3例发生TPMT*3C杂合突变,其活性为(11.99± 1.32)nmol·g-1Hb·h-1,27例呈野生纯合型,其活性为(24.95±11.32)nmol·g-1Hb·h-1,杂合子突变患儿TPMT活性明显低于野生纯合子型患儿,差异有统计学意义(P＜ 0.05).启动子-100附近区域发现5种数目变异的串联重复(VNTR)基因型(*V3/* V3、*V3/* V4、*V4/* V4、*V5/* V5、*V4/* V6),其平均活性分别为19.35、25.06、22.61、32.16、11.85 nmol·g-1Hb·h-1,各组间活性差异无统计学意义(P =0.186).结论 TPMT基因多态性与其活性有一定的相关性,TPMT活性高低对于儿童ALL预后的判断和指导个体化用药有重要的临床意义.
목적 탐토구기표령갑기전이매(TPMT)활성화기유전다태성재인동급성림파세포백혈병(ALL)개체화치료중적림상의의.방법 운용고효액상색보법(HPLC)대100례초진ALL환인、180명건강인동진행TPMT활성검측.채용재선Primer3연건설계인물,대30례구유림상사건적환인진행TPMT기인DNA측서.분석TPMT활성、유전다태성여림상불량반응적상관성.TPMT활성이재단위시간(1 h)내단위질량적혈홍단백(Hb,g)최화생성적2-안기-6-갑기구기표령(6-MTG)적량(nmol)표시.결과 100례초진ALL환인TPMT활성위(31.72±10.31)nmol·g-1Hb·h-1,180명건강인동TPMT활성위(30.70±9.67)nmol·g-1Hb· h-1,량자TPMT활성차이무통계학의의(P=0.450).19례발생엄중골수억제적환인TPMT활성위(20.96±7.24) nmol·g-1Hb·h-1,6례ALL복발자초진시TPMT활성위(40.46±8.18) nmol·g-1 Hb·h-1,량자분별여30례구유림상사건적ALL환인초진시활성[(24.07±11.43)nmol·g-1Hb·h-1]상비차이구유통계학의의(P치균＜0.05).5례화료후발생엄중간장독성자초진시TPMT활성위(23.60±7.48) nmol·g-1Hb·h-1,여30례ALL환인초진시활성상비차이무통계학의의(P =0.930).대30례구유림상사건적ALL환인TPMT기인DNA측서,3례발생TPMT*3C잡합돌변,기활성위(11.99± 1.32)nmol·g-1Hb·h-1,27례정야생순합형,기활성위(24.95±11.32)nmol·g-1Hb·h-1,잡합자돌변환인TPMT활성명현저우야생순합자형환인,차이유통계학의의(P＜ 0.05).계동자-100부근구역발현5충수목변이적천련중복(VNTR)기인형(*V3/* V3、*V3/* V4、*V4/* V4、*V5/* V5、*V4/* V6),기평균활성분별위19.35、25.06、22.61、32.16、11.85 nmol·g-1Hb·h-1,각조간활성차이무통계학의의(P =0.186).결론 TPMT기인다태성여기활성유일정적상관성,TPMT활성고저대우인동ALL예후적판단화지도개체화용약유중요적림상의의.
Objective To investigate thiopurine S-methyltransferase (TPMT) activity and gene promoter polymorphism to probe its significance of individual chemotherapy in acute lymphoblastic leukemia (ALL) children.Methods The average TPMT activities were (31.72 ± 10.31) nmol · g-1Hb · h-1 and (30.70 ± 9.67) nmol · g-1 Hb · h-1 in ALL and healthy groups respectively,without gender differences of TPMT activities (P =0.45) in both groups.The TPMT activity with clinical events in newly diagnosed ALL patients (n =30) was (24.07 ± 11.43) nmol · g-1Hb · h-1.There are significant differences of TPMT activities between severe bone marrow suppression [(20.96 ± 7.24) nmol · g-1 Hb · h-1] and ALL patients with clinical events groups (P ＜0.05).The TPMT activity of(40.46 ±8.18) nmol · g-1Hb · h-1 in recurrence children was also significantly different (P ＜ 0.05).TPMT activity in severe liver toxicity group was not significantly different (P =0.930).Of TPMT gene sequencing in ALL patients with clinical events,only 3 children were heterozygosity mutations of TPMT * 3C,while others homozygous genotype.There were significant differences of TPMT activities between heterozygosity genotype [(11.99 ± 1.32) nmol · g-1 Hb ·h-1] and homozygous genotype groups [(24.95 ± 11.32) nmol · g-1 Hb · h-1] (P ＜ 0.05).There were five kinds of variations at the vicinity of the promoter region of-100 of tandem repeats (VNTR) polymorphism(* V3/* V3、* V3/ * V4、* V4/* V4、* VS/* V5、* V4/* V6) without significant differences of TPMT activities among five kinds (P =0.186).Conclusion TPMT activity was related to the gene polymorphism.TPMT activity determination had prognostic value and guided individualized treatment.