Y HQC Vl|T NAM THANG 8 • Stf BJiC 8161/21112 song, NXB Khoa hgc X ! hgi vi Nhan vSn.
Nguyin Th| Mai (2011), Luin in Tiln st Y hgc, Nghiin dot rii lopn trim .cam to dud tre em bf ung thu vi ing phd cua cha me. Dji hgc Y Hi Ndi.
Nguyin Sinh Phic (2007), Tim thin hpc vi (am 1^ Apc^ AtK, NXB Quin dOi nhin din.
Nguyin Thj Doan Tnmg (2008), Khda lu|n tit nghiep, Bute diu nghiin ciu rii nhiiu dm xic
(loiu- trim cam) a tre em bi bfnh ung thu tpl khoa Ung budu -BVNhi Trung uang, Tnidng DH Khoa hpc Xi hdi vi Nhin vSn.
A.VXavaxeilc6, Iu.G.Dubarep, (I991X Tim ly hpc binh nhdn,YiXBYi«>c.
Lovibond, S. H., & Lovibond, P. F, (199S), Manual for the Depression Anxiety Swess Scales, Psychology Foundation of Australia.
IJMG DUNG Kf THUAT FISH KHAO SAT CHUYEN DOAN T(1;19) TRONG BfNH BACH cAu cAP BONG LYMPHO
TOM TAT
Chuyen doan t(l;19)(q23;pl3) l i y£u td hin ludng xau, gap trong 3 - 5 % benh b^ch c3u cip ddng lympho B. Cd 2 kleu chuyen doan t(l;19) gom chuyen doan can bing va chuyen doan khdng can bang, trong dd nhdm chuyen doan khdng cin bing chl€m ti le d€n 75% v i dugc cho li cd den lUdng tot hdn. Tir thing 1/2011 den thing 4/2012, chung toi sif dung ky thuat RSH de khio sit kigu chuyen do?n t(l;19) frSn 9 binh nhin cd bleu hien E2A/PBX1 bing ky thuat PCR tai Benh vien TruySn miu Huyet hgc.
Ket qua ciia nghien cdu cho thay 44,4% benh nhin cd chuyen doan t(l;19) cin liSng v i 55,6%
benh nhan cd chuyen do?n t(l;19) khdng cin bang, trong do 1 benh nhan cd chuyin doan t(l;19) cin bing nhuhg cd them 1 tin hieu NST 1. Ky thuat FISH la ky thuat ddn giin, cho ket
Le Nguyen Kim Dung*, Pban Th) Xinh*
qui nhanh v i vdi doan dd die hieu gliip xic djnh chinh xic cic kilu bat thudng cila t(l;19).
SUMMARY
DETECTION OF t(l;19) TRANSLOCATION IN ACUTE LYMPHOBLASnC LEUKEMIA USING FISH
TECHNIQUE
The translocation of t(l;19)(q23;pl3) Is a poor prognostic factor, occurs in 3 - 5% of patients with B-cell acute lymphoblastic leukemia.
There are two fomis of t(l;19) translocation Including balanced t(l;19) and unbalanced der(19)t(l;19), in there the unbalanced rates up to 75% and expected to be a better prognostic.
From January 2011 to April 2012, fluorescent in situ hybridization (FISH) technique was used to detect t(l;19) b-anslocatlon of 9 patients with
• Khoa Dl truyen hgc Phan Tu; Binh vien TruySn mau-Huy&hpc TP.HCM.
Phan Ufn khoa hpc: TS. Phan Thj Xinh
137
CHUVeN eg; HOI NOHI KHOA HQC HUYBT HQC • TRUYJN MAU TOAN QU6C 2012 expression of B2A/PBX1 by PCR technique In the
Blood Transfusion and Hematology hospital.
Results of the study showed that 44.4% of patients with balanced t(l;19) and 55.6% of patients with unbalanced der(19)t(l;19). In which a patients with balanced t(l;19) but there Is an additional signal chromosome 1. FISH technique Is technically simple, results In fast and with specific probes help accurately Identify abnomiallty patterns of t(l;19).
[. oijiT V £ N O I
Bfich ciu d p ddng lympho (BCCDL) li mOt rii l o ^ ic tinh cda tl bio diu ddng lympho B hoic T, die tnmg bdi sy ting sinh vi tich ty tl bio non trong tiy xucmg giy ra suy giim cic ddng te bio miu binh thudng [1]. Benh BCCDL chiim khoing 80% ti-ong tit ci cic ca b^nh bach ciu d p d tri em vi khoing 20% cic trudng hgp d ngudi Idn [5].
Chuyen doan nhiem sic the (NST) li yiu ti ti& lugng quan trpng trong BCCDL. Mgt sd chuyin doan NST tao ra nhiing to hgp gien die himg trong benh BCCDL nhu t(9;22) (q34;qll), t(l;19) (q23; pl3), t(4;ll) (q21;q23) va t(12;21) (pl3;q22) tao ra to hgp gien BCR/ABL, E2A/PBX1, MLUAF4 va TEUAMLI tuong ling [1]. Benh nhin cd t(12;21) thi thugc nhdm tidn lugng tdt, ngugc lai binh nhan cd mdt trong 3 chuyin dogn NST cdn lai thi thugc nhdm tien lugng xau.
Chuyin dosm t(l;19)(q23;pl3) xiy ra ti-ong khoing 5% tri em vi 3% d ngudi Idn bi benh BCCDL-B [1]. Cic phin tich vl di truyin hgc tl bio di xac dinh dugc hai dang cua chuyin doan t(l;19) gom chuyen dogn can bing chiim 25% cic trudng hgp, vi phd bien ban li chuyin doan khdng can bing chiim khoang 75% [3,8]. Chuyin doan t(l;19) tjo ra sy td hgp ciia exon 13 cua E2A (cdn dugc ggi la TCF3) vdi exon 2 cua PBXl
dl ding phit h i ^ bing k$> thu$t k h u i c h ^ gien sao mi ngugc (RT-PCR). Tuy nhi&i, § thujt RT-PCR khdng phan bi|t dugc 2 kilu bit thudng ciia t( 1; 19). Vi viy, chiing tii li dyng ky thuit FISH khio sit chuyin ttt^
t(l;19) nhim dinh gii cic kieu bit thuong cia t(l ;19) ttin b ^ nhin BCCDL-B tai Benh vijn Truyin miu Huyit hgc TP.Hi Chi Minh.
II. etfI TUpNG vA PHUCMG PHAP N G H I N CDU 2.L s i i tiling:
Nghiin ciiu dugc tiln hinh tifn 9 benh nhin BCCDL-B cd biiu hi?n t i hgp gien E2A/PBX1 bing ky thu^t RT-PCR t?i Bfnh vien Truyin miu Huyit hgc TP.HCM ti thing 1/2011 din thing 4/2012.
2.2. Phuvng phip nghiin cAu 2.2.1. Thtilkinghiin cAu:
Md ti lo^t ca
2.2.2. Phinmg phip tiin hinh Benh phim li 1-2 mL miu tuy xuong ching ddng ACD hoic heparin cua benh nhm BCCDL-B duong tinh vdi ti hgp gien E2A/PBX1. Mgt phin mSu dugc diing dl liy bgch d u vi ci djnh tl bio trin tiiu bin, phiu cdn 1^ dugc nudi d y trong mdi tnidng RPMl 1640 dl phin tich nhiem sic thi. Doan do dugc su dyng de lai hda li LSI TCF3/PBX!
dual color, dual fusion translocation probe (Vysis, Hoa Ky) cd d u tnic nhu hinh 1. Ti6i bin tl bao vi do?n dd din?c biin tinh bing miy ThermoBrite (Abbott, Hoa Ky) d 75°C hong 5 phiit, sau dd u d 37°C qua dim (khoing W18 dln20tilngdinghd).
Sau khi il, tiiu bin dugc nia dl loai bo nhOng mau dd bit cip khdng die hieu Hng dung dich 0,3% NP-40 / 0,4 x SSC d nhiS dp 75°C trong 2 phut, ke tiip nia bing dung djch 0.1 % NP-40 / 2 X SSC d nhiet do phdng tion^:
phiit. Tiiu bin dugc dl khd vi phu bing 20 jiL dung djch DAPI II counterstain (Vysis) tni*
138
Y HQC VI$T NAM THANG 8 - S 6 BAC BI$T/201 2
khi quan sit dudi kinh hiin vi huynh quang.
u
LSI PBXl I9p13.3n*a)0n
B
Hittk I: cdu trtic Oo^n dd E2A/PBX1 (E2A cdn Oifffc gpi tin M TCF3) Bogn dd tgi vung lq23 phu toan b$ gien PBXl trdi ddi 635kb gdn chAtphdt huynh quang mdu do (A) vd dogn dd tgi vung I9pI3.3 trdi dai 730kb phu gien E2A vd cdc gien a ddu 5' vd
3' cda E2A. gdn chdtphdt huynh quang mdu xanh (B).
Mm ten chi vf tri diem gdy.
III. KiT QUA
De xac djnh ket qua, dgc kieu hinh tin hieu huynh quang phat ra a 200 te bSo tr§n tieu ban cua moi benh nhan dirgrc phan tich. Trong nghien curu nay, chung toi chpn 9 b$nh nhan co ket quii duong tinh t6 hgip gien E2A/PBX1 bing k^ thu^t RT-PCR trong tong so 104 b?nh nhan BCCDL-B (8,7%), dl khdo sat c^c kieu h\h\x hi?n bang ky thuat FISH. Ket qua duQC trinh bay a bang 1.
ST T 1 2 3 4 5 6
Nam sinh 2007 2003 1997 2006 2008 2009
Gidi tinh Nam Nam Nam NO' Nam Nam
Bang 1: K i t q u i FISH v i NST dd cua benh nhan BCCDL KSt qua FISH
52% te bao cd t(l;19) vdi 2 tin hieu lq23, 1 tin hieu 19P13.3 va 2 tin hieu to hop qiula NST 1 v i 19 88% te bao cd t(l;19) vdi 2 tin hieu lq23, 1 tin hieu 19013.3 va 1 tin hieu td hOo qitia NST 1 va 19 88% te bao cd t(l;19) vdi 2 tin hieu lq23,1 tin hieu 19D13.3 va 1 tin hieu to hdp qiiis NST 1 v i 19 62% te bao cd t(l;19) vdi 2 tin hieu lq23,1 b'n hieu 19P13.3 v i 1 tin hieu to hop qiiJa NST 1 va 19 90.5% te bao cd t(l;19) vdi 2 tin hieu lq23,1 tin hieu 19P13.3 va 1 tin hiiu td hdp qiuei NST 1 v i 19 98% te bao cd t(l;19) vdi 2 tin hieu lq23, 1 tin hieu
Ket qua NST do Khdng cay Khdng cay Khong cay Cay khdng moc 46,XY,der(19)t(lil9)(q
23;pl3)ri61/46,XYr41 46,XY,der(19)t(l;19)(q 139
C H U Y S N Bi: HOI N G H | KHOA HQC H U Y 6 T HQC • T R U Y S N M A U T O A N QUgc 2012
7 8 9
1995 2005 2010
NO' ND' Nam
19013.3 vi 1 tin hl$u t5 hdp qiOa NST 1 vi 19 89% tS bio cd t(l;19)(q23;pl3.3) chuSn 94% t£ bio cd t(l;19)(q23;pl3.3) chujin
32% t i bio cd t(l;19)(q23;pl3.3) chuSn
23;pl3)ri71/46.XY[31_
Khdng ciy Khdng ciy 46,XY,t(l;19)((|23
;pl3)ri2]/46,XYr81
l : u l ( l ) 2: iil(19) 3,4: t(l;19)
B ilJl
4|.J«_J^R«.jp_j|3__i«_
Hinh 2: Kiiu chuyin dopn I(l;19)(q23;pl3.3) cin bing
Kit qua FISH cho thiy cd 1 tin hifu NST 1. I tin hifu NST 19va2 tin hifu td hpp (A), luang img vdi I NST 1, I NST 19 binh thuang vi 1 der(l). 1 der(19) do 1(1:19) tpo ra (B>
Hiu hit cic tnrdng hgp ii tri em, chi cd 1 benh nhin 17 tuoi vi benh nhin nam chiim da so (6 trong 9 frudng hgp, 66,7%). Trong 9 benh nhin chi cd 3 benh nhin cd kit qui NST d6 de so sinh, 1 benh nhan ciy khdng mgc va 4 benh nhan cdn l?i khdng lim NST dd. Ket qui FISH cho thiy cd 2 kilu biiu hien ciia t(l;19) gdm kilu chuyin do?n can bing (chuin) v4 kieu chuyen doan khdng cin bing (Bing 1). Chuyin do?n t( 1; 19) can bing gip trong 4 Bitdng hgp (44,4%), tao ra 1 tin hieu mau do (NST lq23), 1 tin hieu miu xanh (NST 19pl3.3)vi2 tin hieu miu vang ciia der(l) va der(19) (Hinh 2A). Kit qua niy tuong img vdi 1 NST 1 va 1 NST 19 binh thudng, vi 2 tin hieu td hgp li cua der(l), der(19) tren NST dd (Hinh 2B). B$nli nhan 1 cd t(l;19) cin bing nhung cd them bit thudng cua NST lq23 (Bing 1).
140
Y HQC VigT NAM THANB 8 • s6 CAC BI6T/2012
i r 14 IS 16 17 18
r £ •> -A?--M- i
flin* J: ificK CAHVCTI dopn I(l;19)(q23;pl3.3) khdng can bing Kit qua FISH cho thiy cd 2 tin hifu NST 1, I tin hifu NST 19 vai tin hieu td hgp (A),
tuong ling vdi kit qud NST dd gdm 2 NST 1 binh thuang, I NST 19 binh thuang va 1 NST 19 do 1(1:19) tao ra (B).
Kilu chuyin doan khdng cin bing biiu hien gdm 2 tin hieu miu do (NST lq23), 1 tin hieu miu xanh (NST 19pl3.3) va 1 tin hieu td hgp mau ving (Hinh 3A) gip trong 5 tirudng hgp, chiim ti le 55,6%. Kit qua FISH tuong img vdi kit qui NST dd vdi 2 NST 1 binh thudng, 1 NST 19 binh thudng vi chi cd 1 der(19) mi khdng tim thiy der(l) (Hinh 3B).
IV. BAN LUAN
Mgt si nghien ciru trudc diy bio cao ring nhdm benh nhan BCCDL-B cd chuyin doan t(l;19) cin bing se cd tien lugng xiu hon nhdm cd t(l;19) khdng can bing [7,9].
Tuy nhien, cic nghien ciru trong nhiing nim gin diy cho thiy khdng khac nhau vl thcri gian sdng khdng biin ci va thdi gian sdng toan thi giiia 2 nhdm [2,4]. Diiu niy cd thi do cac nghien ciru da sir dung phac dd diiu tii BCCDL-B khic nhau va cac phac do
trong thdi gian gin day cd the cii thien dugc tien lugng cua nhdm t(l;19) cin bing. Kit qui nghien cuu cua chung tdi cho thay 44,4%
cd t(l;19) can bing vi 55,6% la chuyin do?n khdng can bing, tuong ducmg vdi cac nghien cim khic [4,7]. Benh nhan 1 (Bang 1) c6 t(l;19) can bing nhung cd them 1 tin hieu NST lq23, nhu vay benh nhan cd thi cd ttisomy 1 vi t(l;19) xay ra ti-en 1 NST 1, tuy nhien der(l) khong bj mit di nhu trudng hgp chuyen do?in khdng cin bang [6].
CHUYEN e g : H Q I N Q H I K H O A HQC HUYfiT HQC • TRUYJN M A U T O A N Q U 6 C 2012
V. KST LUAN
K^ thu§t FISH giup nh$n di$n nhanh c6c ki^u b^t thuimg cOa t( 1; 19) m6 khfing c4n thu dugc te bAo phfin chia. Can c6 nghifin cihi tr£n so mhx \(m hon vk thbi gian kh^o sdt ddi hon de ddnh gid ti€n lui^ng ciia 2 nhdm BCCDL-B c6 t{l;19) cdn b&ng vd khdng cfin bang tai Vi?t Nam.
TAI Llfu THAM KHAO
1. Phan Nguyin Thanh V i n , Phan Thf Xinh, Triln Thj T r i e Thanh, Nguyin TAn Bfnh, "Bdt thurfmg nhiSm sSc th^ vA gen trong b$nh b^ch ciu dp ddng lymphfl". Y h<?c TP.HCM 2011;
15(4):72-78.
2. Andersen M.K, Autio K, Barbany G, el ai
"Paediatric B-cell precursor acute lymphoblastic leukaemia with t(l;19Kq23;pl3): clinical and cytogenetic characteristics of 47 cases from the Noniic countries treated according to NOPHO protocols". Br J Haematol 20\ 1 Oct;155(2):235-43.
3. Brandon M.S, Heather C F , Ryan A.K, et al.
"Interphase FISH to detect PBX1/E2A fusion resulting from the der{19)t(l;19)(q23;pl3.3) or t(l;l9){q23;pl3.3) in paediatric patients with acute lymphoblastic leukaemia". British Journal of Haematology 2005A29: 45-52.
4. Felice M.S, Gallego M.S, Alonso C.N, et al.
"Prognostic impact of t(l;19y TCF3-PBXI in
childhood acute lymphoblastic leukemia in the context of Berlin-Frankftirt-MUnster-hised protocols". LeukLymphoma20\ \ Jul;52{7);1215.2l, Mesqulla D.R, Cdrdoba J.C, Magalh8estQ,a at. "Molecular and chromosomal mutations among children with B-lineage lymphoblaitic leukemia in Brazil's Federal District". GeneAa and Molecular Research 2009;8{ 1): 345-353.
Paulsson K, Horval A, Fioretos T, el d.
"Formation o f der(19)t(I;19Xq23;pI3) in acute lymphoblastic leukemia". (Jene.'i Chromosoma Cancer 2005 Feb;42(2); 144-8.
Seeker-Walker L.M, Berger R, Fenam ?,elal
"Prognostic signiflcance o f the balanced t(l;19) and unbalanced der<l9)t(l:l9) translocatiooi Ji acute lymphoblastic leukemia". Leukemia 1992 May;6(5):363-9.
Uckun F.M, Downing J.R, Gunlher R,ad
"Human t(l;l9Xq23;pl3) pre-B acute lymphoblastic leukemia in mice with sevat combined immunodeficiency". Blood 1993;ll (81): 3052-3062.
Uckun F.M, Nachman J.B, Satber H.N, etd
"Clinical significance o f Philadelphia chromosome positive pediatric acute lymphoblastic leukemia in the context of contemporary intensive therapies; a report from the Children's Cancer Group". Cancer 1998 Nov 1 ;83(9):2030-9.
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