Tgp chi Cong nghe Sinh hoc \\{\): 27-32, 2013
N G H I E N C U U G E N CYPIIB2 6 N G U O I B E N H C O SU T.ANG C A O A L D O S T E R O N E Ngu\en Huy Hoiing'•^ Rita Bernhardt^
' Vien Nghien ciru he gen, Vien Hdn ldm Khoa hoc vd Cong nghe Viel Nam 'Dgi hpc Tdng hop Saarland, Cong hoa lien bang Dire
TOM T A T
Aldosterone la thanh phan chinh cua nhom mineralocomcoid hen quan ten sy dieu hoa muoi va niroc trong CO the Khi aldosterone tang cao se gay ra sir tang cao huyel ap trong mau Trong nghien cuu nay, chiing toi nghien ciru benh nhan nam co bieu hien tang cao aldosterone va tieu dircmg typ 2 Benh nhan nay duoc chan doan CO hien tu'png glucocorticoid ap che tang cao aldosterone (GRA) GRA xay ra do nguyen nhan cua qua irinh trao d6i cheo giua gen CYPIIBI va CYPUBZ Toan bo gen CYP1IB2 (khoang 7 kb) va viing Promoter ciia gen CYPlIB2 (khoang 2,2 kb) duoc nhan len va giai trinh tu gen, Oanh gia cac bien dot da hinh tren timg allele cua promoter gen CYPlIB2 thong qua viec chon dong san pham PCR vao vector pGow-TOPO va giai trinh tir gen tir cac dong thu duoc, Ket qua cho thay chi c6 mot dot bien da hinh tai vi tri R374K tren gen CYPl IB2; Khong co sir Irao doi cheo Iao gen ao" dau 5" mang gen CYPIIBI va dau 3" mang gen CYPIIB2.
Da phat hien duoe 13 diem bien doi da hinh iren promoter ciia gen CYPl IB2 lai cac vi tri -84, -130 den -131. - 257, -337.^64, -638,-656,-695, -1415 den -1418, -1506,-1577 d^n -1579.-1644 va-1740 Nhirng bicn doi da hinh nay co Ihe la nguyen nhan chinh gay anh hudng tdi su lang cao aldosterone d nguoi benh.
Tir khda: Aldosterone, CYP1IB2, CYPl IB2 pro luio doi ilico giira gen CYPIIBI vd CYPIIBI
r. glue irlicoid dp che tdng cao aldosterone lGR.4).
MO DAU
Aldosterone la thanh phan chinh ciia nhom mineralocorlicoid, duoc bieu hien d viing glomcrulosa va duac dieu khien chinh bdi angiolensin II va kali trong mau. Tren \ o thirong than, su tong hop aldosterone dupc xiic tac bdi mot so enzyme cytochrome P450 (Curnow el al, 1991).
O ngudi, budc cuoi ciing cho qua trinh long hop aldosterone duoc xiic tac hot enzyme CYP11B2 CYP11B2 xiic tac qua trinh chuyen hoa deoxycorlicoslerone(DOC) tdi aldosterone, qua trinh nay bao gom ba budc hydroxyl hoa tai vi tri 1 ip de lao ra corticosterone (B), tiep iheo lai vi tri 18 de tao ra IS-hydroxycorticosterone ( I 8 0 H B ) , va cuoi ciing la 0X1 hoa lai vi tri 18 de tao ra aldosterone (Bureik et al., 2002).
Hpi chimg lang aldosterone so cap dugc bieu hi<;n ciia tSng cao huyet ap ciing vai sy du thira aldosterone, mk muoi kali. \ a ap che he thong rennin-angiolensin Thong thudng. bicu hien phu lam Siing cua lang aldosterone so cap la do san pham aldosterone ciia u tuyen thuong than \ a lang san ihu^mg Ihan d 2 iu\cn vo. Glucocomcoid ap che liing aldosterone (GR-A) rat il gap trong tang aldosterone so cdp. GRA dac trung bai su bai net
aldosterone tdi adrenocorticotropic hormone (ACTH) khong binh thudng va no chiem uu the hon vdi he ihong rennin-angiotensin. trong su dieu khien cua bai tiet aldoslerone (Gordon. 1994)
Gen mii hoa CYP11B2 nam tren chromosome 8q22, CO kich thudc khoang 7 kb bao gom 9 exon va 8 inlron. Gen CYPIIBI co do tuong dong idi 95% doi vdi viing exon va 90% doi vdi vimg intron so vdi gen CYPIIBI Diem khac biel dang ke ve ciiu triic cua hai gen CYPIIBI va CYPltB2 la them mot doan 442 bp trong inlron 5 cua CYP11B2 (Kawamoto el al. 1992.
Momet et ai. 1989, Brand el at., 1998) Cac dot bi^n tren Ol'l IB2 la nguyen nhan chinh cua viec suy giam aldoslerone (Nguyen el al, 2008; Nguyen el cd., 2010.
Lovas (*/ cd.. 2009) Cac nghien cuu gan day cho thay gen ao CYPIIBI/CYP1IB2 la nguyen nhan cua GRA (Lifton el ai. 1992, Lifton el ai. 1992, Pascoe et at.
1992; Miyahara el ai, 1992) Gen ao nay co dau 5' la CYPIIBI va ph^n phia dAu 3" la CYPIIB2, do vay su bieu hien protein hen quan tdi long hirp aldosterone dudi su dieu khien ciia promoter cua gen CYPIIBI Ngoai ra. Hampf\ac6ng su(20()l) da mim Ui gen a o a ngudi benh cb hien tuong tang san ihucmg than bam sinh vdi cdu true diiu 5' la C)I'IIB2 dau 3" la CYPIIBI \ a llic nay promoter cua gen ( ) PI 182 dieu khien qua trinh long hop Cortisol
Trong nghien cuu nay, chiing toi tien hanh nghien ciru gen d benh nhan co hien tugng tang cao aldosterone d ngudi benh tieu dudng typ 2. Phan tich kha nang trao d6i cheo giua 2 gen CYPIIBI va CYPI1B2 a benh nhan, va ti^n hanh sang loc dot biSn tren gen CYPIIBI va promoter ctia gen CYPIIB2.
NGUYEN LIEU VA PHU'ONG PHAP Nguyen lieu
Cac hoa chat thong dyng dugc sii dung la san pham ciia hang New England Bioiabs, Merck/BHD Chemicals (Due), Sigma (My), .pGlow-TOPO vector (Invilrogen).
Benh n h a n
Benh nhan nam nhap vien tai Phong kham da
Nguyin Huy Hoang & Rita Bernhardt khoa. Benh vien Trudng Dai hpc Eriangen, CHLB Diic. Benh nhan co hien tugng tieu dudng type 2.
Dac biet, k6t qua xet nghiem hda sinh cho thay str tang cao aldosterone la 475 pg/ml so vdi ngudi binh thudng la 30-160 pg/ml. Chan doan benh nhan co hien tugng glucocorticoid ap ch^ tang aldosterone, Nhan vd gidi trinh tie gen
DNA tdng s6 ciia benh nhan dugc tach chiet theo phuang phap ctia Sambrook va cong sy (1989) d6 tao khuon mau cho phan ung nhan gen PCR.
Toan bg gen CYP11B2 bao gom tit ca exon va intron duoc nhan bang 3 cap moi rieng biet theo theo Nguyen va cong su (2010). Promoter ctia gen CYPIIB2 dugc nhan len vdi cap moi AP-F, AP-Rva dugc dgc trinh tu gen vdi moi A I - A 6 (Bang I) Ket qua dgc trinh Ur duoc phan tich va so sanh vdi trinh tu promoter cua gen CYPIIBI chuan trong ngan hang gen bang phan mem Bio Edit 3 0
Bang 1: Ooan oligonucleotides
Trinh t y (5' > 3') AP-F
AP-R
GAATTCTGCATCCTGTGAAATTATCCTTCA CAGCTCCTGGAAGGTCTGGTGCATC ATCTGCAGCAGCCTCAGCCA GGGTCTGCTCCTGGTGGTCAACATGCT GCGGTGACTCACCACACCTGTAATCC CAGCAATGATGCAAGGGATGGGTGAGTGGA TCCACTCACCCATCGCTTGCATCATTGCTG GTGGAAGGTGGTGGGACCTGGCCT CCCATGACGTGATCCCTCTCGAAGG AAACACAGGCCCTGACCCGTATCCC GAGCGTCATCAGCAACGGAAACGCT
Phan img PCR duoc tien hanh vdi long the tich la 50MI bao gdm IX Buffer, 10 inM dNTP, 10 pmol mOi mm. 200 ng DNA long s6 va 4 U BIO-X-ACT DNA polymerase (Bioline) Chu trinh nhiet nhan promolei cua gen CYPIIB2 bat dau bang lien bien tinh 9?"C Uong 5 phiit. 60"C trong I phiit tiep theo la 30 chu ki gan moi \ a keo d.ii bao gom keo dai chuoi a 68"C trong 5 phtit. bien tinh 95"C trong 30 giay, va gan moi a dO'T trong 30 giay, budc cuoi ciing la 6S"C trong 10 phiit. San phiim PCR sau do duac tinh sach hang bo Kit Gel Extraction cua hang Fermcnias Phan ung PCR danh gui su trao doi cheo giua 2 gen Ol'l IBI Vli CYPIIBI duac ti^n hanh vdi long
the tich 50 ^1 bao gom IX Buffer, 10 mM dNTP, 10 pmol m6i mdi (C-3 va A-6), 20 pmol C - l , 200 ng DNA tOng s6 va 4 U BIO-X-ACT DNA polymerase (Bioline), Chu trinh nhiel nhan kiem Ira sy xuat hien trao doi cheo nhu sau; bi^n tinh 95"C trong 3 phiit;
tiep theo la 30 chu ki gan moi va keo dai bao gom:
bien tinh 95"C trong I phiit, gdn mdi d 60"C trong I phut va keo dai chuoi o 72"C trong 5 phiit; budc cuoi cimg la 72"C trong 10 phiil
Chan ddng gen trong vector pGlow-TOPO Vector pGlow-TOPO duac thi^t k6 vdi Irinh tir phia trudc co gan doan trinh tu ma hda protein phat
Tgp chi Cong nghe Smh hoc \\(\y. 27-32, 2013 huynh quang xanh-green fluorescent protein (GFP).
Toan bd promoter cua gen CYPl IBI dugc gan vao vector pGlow-TOPO, sau do duac kiem tra bang dgc trinh ty gen su dung moi Al va A4.
KET QUA VA THAO LUAN
Ket qua nhan va dpc trinh ty gen CYP11B2 (kich thudc khoang 7kb) d ngudi benh co hien tugng tang cao aldosterone cho thay: c6 mgt dot bien di hop tu nucleotide tai vi tri 518 tren cDNA bien doi nucleotide tii G thanh A, dot bien nay dan den su thay the arginine thanh lysine tai vi tri amino acid 173 Tuy nhien cac nghien cuu trudc do da chung minh dot bien nay khong anh hudng tdi hoat tinh enzyme C Y P n B 2 sau khi tien hanh thu hoat tinh tren te bao dpng vat (Nguyen el ai, 2008; Portrat- Doyen etai, i998)
Gen ao CYPl IBI/CYPl IB2 duac xem la nguyen nhan ciia GRA (Lifton et al, 1992, Lifton el al, 1992, Pascoe e? a / , 1992; Miyahara e; a/., 1992) Gen ao mang trinh ty dieu khien dau 5' cua gen CYPIIBI va trinh tu dSu 3 ' ciia gen CYP1JB2, do do sy bieu hien protein vdi hoat tinh tong hpp aldosteone dugc dieu khien dudi promoter ciia gen CYPIIBI va ACTH (Hinh 1 A) Cac dilm trao d6i cheo nay nam tren intron 2 den intron 4 da duoc tim
*h5y d ngudi benh GRA (Lifton el ai, 1992; Pascoe
el ai, 1992). Ket qua nghien cuu sy trao doi cheo giira 2 gen CYPIIBI vi CYPIIBI bSng each su dung 3 cap moi dac hieu trong ciing mpt phan img (Hinh 1), cho thay mau ngudi benh chi co mot bang duy nhat tucmg ung vdi mau doi chimg. Trong trudng hgp co sy trao doi cheo gen CYPIIBI va CYPl 182, vdi viec su dung 3 moi dac hieu se cho ra 2 bang 4,4 kb (Hinh LA) va 1 bang co kfch thudc 2,7 kb (Hinh 1 B) Tuy nhien ket qua phan iing PCR d ngudi benh nay chi cho ra mgt bang duy nhat co kich thudc 2,7 kb, chung to khong co hien tuong trao d6i cheo giua 2 gen CYPIIBI va CYPIIBI
O vo thucmg than va tuyen smh due ciia ngudi trudng thanh, steroidogenic factor I (SF-1) la yeu to chinh cho viec bieu hien ciia cac gen hydroxylase steroid bao g6m CYPl IAI (Clemens e! al, 1994.
Liu and Simpson, 1997), 1 ip-hydroxylase ( C Y P I I B I ) (Wang el ai, 2000), 21-hydroxylase (CYP21) (Ikeda el ai, 1993), 17a- hydroxylase/17,20 lyase (CYP17) (Givens el at, 1994) va aromatase (CYP19) (Michael el al.
1995), Cac viing yeu to cis. Ad tren promoter ciia gen CYPllB co the xem la viing gan vdi SF-I Ngoai ra yeu to CRE/Adl tren promoter cua CYPllB dong vai tro quan trong Irong qua trinh phien ma ciia gen CYPllB (Ye el ai. 2009), Chinh vi vay nhirng bien doi da hinh tai vimg nay cd the se anh hudng tdi su tong hap aldoslerone.
1 A
ti
Promoter CYP11B1
C-1
II
c-1 27
..—
1 m 1 CYP11B2
1
C-6
J[ ^ i 1^
Promoter aen CYPIIBI
H i n h i A So do Sl,!'Irao doi Cheo giQ'a promoler cua gen C V P I J B I v a C y P r 7 e 2 B Ntian So^n gen CY11B1 (6ai chwng) C San pham PCR sir dgng 3 cap m6i C 1 , C3 va_C6 duac phan tach tren gel agarose 1 % , siJ' dung 810-X-ACT DNA polymerase M marker 1 kb, 1 mSu doi chiing, 2 m i u ngu-o'i b^nh
Nguyen Huy Hoang & Rita Bernhardt
c;
P C R product ( - 2 2 kb) of C Y P l 1E2 promoter
<=
- 3 3 7 T/C - 2 S 7 A
^
C Y P 1 1 B 2 p r o
Hinh 2. Sa d6 nhan doan promoter cOa gen CYP11B2va vi Iri cac diem da hinh d u o c phat hien 13 diem bien doi da hinh tren promoter ciia gen CYP11B2
Trong qua Irinh nghien ciru promoter cua gen CYPIIB2, chung toi lien hanh nhan va giai trinh Ur doan promoter ctia gen CYPIIBI, ket qua cho thay da phat hien duoc 13 diem bien doi da hinh bao gom mai, them va thay ihe nucleotide (Hinh 2), De CO the bict dugc dot bien do xay ra tren lung allele, chiing toi tien hanh chon dong trong
vector pGlow-TOPO Doan gen promoter CYPl 1B2 dugc gan vao vector pGlow-TOPO (Hinh 3), sau do dugc bien nap vao te bao E. coh TOP I OF, Mudi dong te bao dugc chpn de tach chiet DNA plasmid va giai trinh ty gen, ket qua cho thay cac bien dot da hinh tren promoter ciia gen CYPIIBI tren timg allele (Bang 2).
Hmh 3. Vector bieu hien promoter ciia gen CYP1 -82
Tgp chi Cong nghe Sinh hpc \\{\): 27-32. 2013
Bang 2. Vjtri da hinh promoter cua gen cyP1tB2\rhn 2 allele Promoter cua gen CYP11B2
Allele -1740 -1644 -1577 -1506 -1415 -656 -638 -464 -337 -257 -130
Allele C 1 Allele C
2 PCR O T
-1579 TGG
• 1418 AAAT
AAAT/- A/-
Chu thich: (-) Mat nucleotide, (+) Them nucleotide, (>) thay doi nucleotide, Y (T+C), W (A+T), R (A+G) Dot bien mat mgt nucleotide d tren I allele xay
ra d VI tri -84, -695, mat 4 nucletide tren I allele tii vj tri -1415 den -1418 va mat 3 nucleolide tren I allele tir vi tri -1577 den vi tri -1579; Dot bien them 1 nucleotide tren 1 allele vao giua vi tri -130 va - 131; Ngoai ra dot bien thay the tren 1 allele tai vi tri -257, -337, -464, -638, -656, -1506, -1644 va dot bi^n thay the tren 2 allele tai vi tri -1740 (Bang 2) Dac biet, VI tri -337 tren promoter ciia gen CYPl IBI (tuong duong vdi vi tri -344 neu tinh vi tri dau tien la ATG) CO anh hudng tdi su gan ciia SF-1 (White el al., 1995) Nhung ngudi CO diem da hinh la-337T se lam tang ty le aldosterone/than va tang bai tiet aldosterone nudc tieu (Davies el al, 1999; Lim el al, 2002). Chinh vi vay nhiing bien doi da hinh tren promoter ciia gen CYPIIBI co the la nguyen nhan gay hien tugng tang cao aldoslerone a ngudi benh.
KET L U A N
Trong nghien cim nay, chiing loi khong phal hien dugc dot bien tren gen CYPl IBI va khong phal hien CO su trao doi cheo giira promoter cua 2 gen CYPIIBI va CYPIIBI d ngudi benh Tuy nhien chiing loi da phal hien duoc 13 bien doi da hinh tren promoter ciia gen CYPIIBI Tit nhung ket qua tren, budc dau chiing toi co the giai Ihich rang sy tang cao aldsoterone ciia ngudi benh co the la do nhimg bien doi Iren promoter ciia gen CYPl IBI.
I.M ciiiii o'n: Chimg ldi ciing xin chdn lluinh cam an Bi'nh vien Tnrang Dai hgc Eriangen, Cpng hda lien bang Dire da cung cdp mdu.
TAI Lli:U THAM KHAO
Brand E, Chatclain N, Mulalero P. et al. (1998) Structural analysis and evaluation of the aldosterone s>Tiihdse gi;iic m hypertcnsiun Hypertension "^2 l4S-2fl4
Bureik tvI, Lisurek M. Bernhardt R (2002) The human steroid hydroxylases CYPIIBI andCYPllB2 Biol Chem 383,1537-1551
Clemens JW, Ula DS. Parker KL, Richards JS (1994) Steroidogenic facior-1 binding and transcnpUonal aLti\it\
of the cholesterol side-chain cleavage promoter m rat granulosa cells Endocrinology \24 1499-150S Cuniow KM. Tusie-Luna MT, Pascoe L, Nalarajaii R. Gu JL, Nadler JL, White PC (1991} The produci of the CYPl 1B2 gene is required for aldosterone biosynthesis m the human adrenal cortex Moi Endocrinol 5- 1513 1522 Davies E. HollowayCD. Ingram MC, Inglis GC, Friel EC, Morrison C. et al (1999) Aldosterone excretion rate and hlood pressure in essential hypertension arc relaled lo polymorphic differences in the aldoslerone synthase gene CYP1IB2 Hypci lension 33 703-7
Gordon RD|i994) Mineralocorlicoid h>T)ei-tensK)u Lancet .144 240 • 24.1
Givens CR. Zhang P, Bair SR. Mellon ,SH (1994) TranscnpUonal regulation of rai cytochrome P450cl7 expression in mouse Leydig M-\-IO and adren.il Y-1 cells idenlilication of a single protein that mediates hoih basal and cAM P-indiiced acm ities D.\'A Cell Biol 13 10X7 -1 ri9S Hampf iM- Dao NT. Hoan NT. Bemhardl R (2000) Ne\^
molecular genetic defects causing I Ihela-hydrox^Lise deliciencylCAH) Endocr Res 26 781-782 Ikeda Y, Lala DS. Luo X, Kim E, Moisan MP Parker KL (1993) Characlenzauon of the mouse f T / - H gene, whiih encodes a key regulator of steroid hydroxylase gene expression Mot Endocrinol 1 852-860
Kawamoio T, Milsuuchi Y. Toda K, et al (1992) Role ol steroid 1 l[}-hydroxylase and steroid 18-hvdro\ylase in the biosynthesis of glucocorticoids and inineraloLoriiciiids in humans Pioc \ad icadSci USi fi9 \4'.X 14W LiOon RP. DhihyRG. Powers M, et ai (19' 1 ip-h\dro\>lase aldosterone synlhase glucocoriicoid-rcmediable aldosteronism hypertension \aiurc 3>5 262 26>
2)-\chmi.i
Nguyin Huy Hoang & Rita Bernhardt Lim PO. Macdonald TM, Holloway C, Fnel E. Anderson
NH. Dov. E, et al (2002) Variation at the aldoslerone synlhase (CYPl 182) locus contributes to hypenension in subjects with a raised aldosterone-to-renin ratio J Ctin Endocrinol Metab ?,! 4398-402.
Liu Z. Simpson ER (1997) Steroidogenic facior-l (SF-1) and SPI are required for regulation of bovine CVTIIA gene expression in bovine luteal cells and adrenal YI cells.
Mot Endoa-inol 11 127-137
Lavas K. McFarlane L Nguyen HH, Curran S, Schwabe J.
Halsall D, Bemliardt R. Wallace AM. Chatterjee VK (2009) A novel CYP11B2 gene muiation in an Asian family with aldoslerone synthase deficiency / Clin Endocrinol Melah 94 914-919
Michael MD, Kilgore MW, Morohashi K. Simpson ER (1995) Ad4 BP/SF-1 regulates cyclic AMP-mduced transcnplion from the proximal promoter (pu) of the human aromata.se p450 (cypl9) gene in Ihe ovary. J Biol Chem 210: 13561-13566.
Miyahara K, Kawamoto T, Mitsuuchi Y, et al (1992) The chimeric gene linked lo glucocorticoid-suppressible hyperaldosteronism encodes a fused P-450 protein possessing aldoslerone synthase activity Biochem Biophvs Res Commun 1S9 885-891
Momet E, Dupont J, Vitek A, White PC (1989) Characterization of two genes encoding human steroid 11 P-hydroxylase {P-45011P) J Biol Chem 264" 20961- 20967
Nguyen HH, Hannemann F, Hartroann MF, Malunowicz EM. Wudy SA, Bernhardt R (2010) Five novel mutations in CYP11B2 gene detected in patients with aldosterone synlhase deficiency type I: Functional charactenzalion and structural analyses. Moi Genet Metab 100. 357-364.
Nguyen HH. Hannemaim F, Hartmann MF, Wudy SA.
Bemhardl R (2008) Aldoslerone synlhase deficiency caused by a homozygous L451F mutation m the CYP11B2 gene. Mot Genet Meiah 93: 458-467
Pascoe L. Cumow KM, Slutsker L, et al, (1992) Glucocorticoid-suppressible hyperaldosteronism results from hybrid genes created by unequal crossovers between CYPIIBI and CYPIIBI. Proc Nad Acad Sci USA 89 8327-8331
Sambrook, Maniaiis T, Fnlisch EF (1989) Molecular cloning: a laboratory manual,
Wang XL, Bassett M, Zhang Y, Yin S. Clyne C, White PC. Rainey WE (2000) Transcriptional regulation of human lip-hydroxylase (^iCXP/ZF/; Endocrinology 141 3587-3594
While PC, Slutsker L (1995) Haplolype analysis of CYP11B2 Endocr Res 2\ 437^42
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1303-1309 STUDY ON ALDOSTERONE
HYPERALDOSTERONISM
SYNTHASE GENE IN PATIENT SUFFERING
Nguyen Huy Hoang'^ ". Rita Bernhardt^
Inslilule of Gen 'Saarland Unive
e Research, Vieinam Academy of Science and Technology y, Germany
,Aldosierone is the main inineraloconicoid in human ll is involved m the regulation of the salt and water hoLiseiiold of the body and thus m the regulation of blood pressure. Herein, we have studied a male patient wilh hyperaldosteronism and diabetes lype 2 The patient was diagnosed with glucocorticoid-re media bie aldosteronism (GRA) GRA IS caused by crossing-over helween CYPIIBI and CYPIIBI gene CYPIIBI gene (approximately 7 khi and C) PI IB: promoter (approximately 2.2 kb) vsere amplified by PCR and sequenced The vanant of on each allele of C) PI IBI promoter gene were performed by CYPl IBI promoter gene cloned into pGlow-TOPO vector, ,iiid ihen 10 clones \\as sequenced The results showed that one mulani R374K was detected in CYPIIBI gene' There was noi chimeric gene cross-o\er helween 5' coding .senquence of CYPl IBI gene and 3' coding sequence oft )PllBI\itne Thirteen'.ariaiionsofCYPllB2 promoter gene were detected in positions'-84 -130to-13l - 257. -331 -4M. -63S -6.%. -695. -1415 lo -1418, -1506. -1577 to -1579, -1644. and -1740 Tlie vanalion of C\ P11B2 promoter could cause affecimi; h\peraldosteronism of patient
hcywords- \ldo--
iriuoid-rcmcduihlcaldosicn.
ween CYPIIBI t
u iGB-i) ul CYPIIBI gene CYPIIBI. CYPl 1B2 promoter.
.•spuiulciKC F-111L1|1 /?.',
