VIETNAM MEDICAL JOURNAL N°2 - APRIL - 2018 - Ty le tre SDD the gay cdm la 5,7%, trong
dd nam la 6 , 1 % , nd la 5,0%.
- Ve ty le tre SDD d cac the tang dan theo nhdm tuot.
KIEN NGHI
Can da dang hda eac loai hinh truyen thdng nang cao ki§n thdc va thUc hanh cham sde tre cho eac ba me ed con dudi 5 tuoi nham cat thien tinh trang dinli dUdng cho tre em tai xa Nghi Van ndi rieng va ngUdi Viet Nam ndi chung.
TAI LIEU THAM KHAO
1. Nguyin Thi Hai Anh, Le Thj Hdp va con^ su*
(2006), "Tinh trang dinh du&ng va mgt so yeu td lien guan den suy dinh dudng tre eni dudi 5 tuoi b'nh Lao Cai nim 2003', Tap chi Dinh Du9ng va ThUc Pham so 3 + 4, thang 11 nam 2006, Hoi nghi khoa hoc Hoi Dinh DUSng Viet Nam lan thir 3 nam 2006.
^. \^^\&i;imi),^Oiudngt3inh phong chong SDD iji em nim 2(X)7, To thU ky ChUdng tnnh Phong chdng Suy dinh du8ng tre em, 48 Tang Bat Ho, Ha Noi.
3. Nguyen Kim Dung (2004), ''Thu'c trang suy dinh duSng d tre dt/di 5 tuoi vi mot so yeu to lien qm den tinh trang do taixa Phung Hutig, huygn Khoi Oliu, Hutig Yen nam 2004", Oe tai thUc dia qiat doan 1^ khoa eao hpc 7, TrUOng Dai hpc Y te Craig corig Ha Noi, nam 2004.
4. Nguyen t h i NhuT Hoa (2011), Tmh tmng mti du&ng vi mot so yeu to lien guan cua tre em dif$
5 tuoi huyen Yen Thuy tinh Hoa Binh, Khoa luan tot nghiep bac sT y khoa, Dai hoc Y Ha Not, 5. Ngo Thj Phu'dng (2007), Thut irang tfmlf
du&ng tre em du&i 5 tudi vi kien thdc, thuth^nli ve cham sdc tre cua bi me tai xa My Hda, huyen Tan Lac, tinh Hda Binh, Luan v3n chuyen khoa I TrUdng,dai hoc y te eong cong
6. Nguyen Thj Vu Thanh (2005), " Thu'c trang va mgt so yeil to inh hu&ng den tmh trang siiydinl) du&ng d tre em dudi 5 tuoi tai xa Sdn Dong, huyin Noil Ddc, Hi Tay, nam 2005'. Luan van tot nghiep Thac sy Y te cong ddng TrUdng Dai hoc Y te Coiig Cgiig nam 2005.
' - Trung tam CSSKSS tinh Nghg An "Bao cao tdng ket cong tac PCSDDTE nam 2016 trien Wiai ke hoach nam 2017".
DANH GIA TAC DUNG KHANG MOT SO DONG TE BAO UNG THU' CtA ALTRETAMIN TONG HO'P TAI VIET NAM
Pham Thi Man', Nguyen Son Nam^ Vu Binh Dirffng'
T 6 M TAT
Muc tieu: nghien euTu tac dung khang te bao ung thu VU(^ ung thU bieu mo phe nang A549, te bao ung thu buong trirng Ovcar-3 ciia Altretamin trong invitro.
Doitddnf va phddng phap nghien cuU: danh gia t^C dung Uc che sU phat trien ciia te bao ung thU ciia altretamin tdnci hpp tren cac dong te bao T-47D, A549, Ovarc bang thU nghiem Trypan Blue. Oanh gia tac dung khang eae dong te bao ung tJiU T-47D, A549, Ovcar-3 ciia altretamin bang thU nghiem MTT. Ket qui: ThU nghiem Trypan Blue, ty le te bao ung thU chet va tdng so te bao d cac gieng nuoi te bao giu^ sir dung chat mi Altretamin tong hpp d eac nong dd 10 pg/ml, 5pg/inl va 1 pg/ml, 0,5pg/ml va Doxombicafi 10 pg/ml khong eo sU khac biet (p>0,05). ThU nghiem WXT eho thay Albetamin tong hpp ire che sU phat trien ciia ^ bao ung thU vu d thdi diem 72 gi6 vdi nong do lO^ig/ml, 5ug/ml (p<0,05); te bao ung thU bieu mo phe nang dong A549 6 thdi diem 24 gid vdi nong dp lOpg/ml, thdi diem 48 gid va 72 gid vd\ cac nong dp lOpg/ml, 5pg/ml va Ipg/ml, 0,5pg/ml (p<0,05). Tac dung gay dut gay DNA te bao ung thU
'B^h vien lao khoa Tmng trddng
^Benh vien TWQuin ddi 108
^Hgc vien Quin y
Chiu trach nhiem chinh: Vu Binh DUdng Email: [email protected] Ngay nhan bai: 17.2.2018 Ngay ph^n bien khoa hoc: 29.3.2018 Ngay duyet bai: 5.4.2018
V(S cac dong vdi cac dong te bao T-47D, A549, Ovarc eiia Altretamin khong cd sU khae biet vdi nhom chimg.
Ket luan: Altretamin co tae dung khang te bao ung thU tUdngjJUdng vdi Doxorubicin; Altretamin ii'c che sU phat trien va tac dung gSy dirt gay DNA te bao ung:
thU vdi cac dong vdi cae dong te bao T-47D, A549;
Ovarc tren thUc nghiSm invitro.
Tdkhda: Altretamin tong hpp, Uc ehS t? bao ung thU tren invitro
SUMMARY
TO EVALUATE RESISTANCE EFFECT WITH SOME CANCER CELLS OF SYNTHESIZED
ALTRETAMINE I N VIET NAM Objeciiires: To study resistance effect of Altretamine with breast cancer cells, AS49 alveolar carcinoma cells, and Ovear-3 cancer cells in invitro.
Subject and method: To evaluate resistance effect of synthesized Altretamine with evolution of T-47D, A549, Ovarc cells by Trypan Blue trial. To evaluate resistance effect of altretamine with T-47D, A549, Ovcar-3 tumor cells by MTT trial. Results: The Trypan Blue trial, the rate of death cancer cells and the total number of cells in the cell culture wells were were not significantly different (p> 0.05) between the synthesized Altretamine reagents with concentrations of lOpg/mt, 5pg/ml and Ipg/mi, 5pg/ml and Doxorubicin lOMg/ml). The MTT trial showed that synthetic altretamine inhibited the evolution of breast cancer cells at 72 hours with concentration of lOpg/ml, 5pg/ml (p <0.05); of A549 alveolar epithelial
40
TAP CHl Y HOC V l i T NAM TAP 465 - THANG 4 - SO 2 - 2018 carcinoma cells at time 24h with concentration of
lOpg/ml, 48h and 72h with concentration of lOpg/ml, 5pg/ml and Ipg/ml, Spg/ml (p<0.05). The effect of cancer cell DNA fracture with T-47D, A549 Ovarc cells, were not significantly different (p>0.05) between the synthesized Altretamine and control group.
Conclusion: Altretamine has anti-eancer cells were equivalent to Doxorubicin; Altretamine inhibited the evolution and cancer cell DNA fracture with T-47D, A549, Ovarc cells in vitro experiments.
Key words: Synthesized Altretamine, inhibit cancer cells in invitro
I. OAT VAN DE
Trong khoang 50 nam trd lai day, ung thU la benh gay tU vong xep thU hai, sau eac benh tim maeh. Hdn 70% tU vong do ung thU la d eac nUdc thu nhap thap va trung binh, ndi cac nguon iUc danh cho phdng chdng, chan doan, dieu tri ung thU edn han che. Theo thdng ke eiia WHO, nam 2016, sd trUdng hdp ung thU mdi (ty le mae benh ung thu) la 454,8 tren 100.000 ngUdi, sd ngUdi ehet vi ung thU la 171,2/100.000 ngUdt.
Ty le tU vong do ung t h u cao hdn d nam gidi so vdi phu nu' (207,9 tren 100.000 nam gidi va 145,4 tren 100.000 phu niJ). Ty le mae ung thU eao nhat d dan dng My gdc Phi (261,5/100.000) va thap nhat d phu ntf chau A - Thai Binh DUdng (91,2/100.000), (sd lieu thdng ke nam 2008- 2012). Sd ngUdt sdng vUdt mire chan doan ung thu da dat gan 14,5 trteu trong nam 2014 va dU kien se tang ien gan 19 trteu vao nam 2024. 6 Viet Nam, nam 2008 cd 111.581 ca mac mdi benh ung thU.
Altretamin la thuoc dUdc sU dung phd bien trong dieu tri ung thU vii, ung thU phoi va ung thu buong trimg. Loai thudc nay hien dang dUde sU dung dieu tri ung thU vdi biet dUde nhir:
Hexastat (Phap), Hexinawas (Tay Ban Nha), Altrenamin (My),..[2]. Cung gidng nhUeac thudc dieu tr| ung thU khac, altretamin hau het phai nhap ngoai vdi gia thanh eao. TU nam 2014, Hpc vien Quan y da nghien cUu thanh cdng quy trinh tdng hdp altretamin [ 1 ] , de san pham ed the dUdc dng dung trong dteu trj thUc te can cd cac nghien ciru ve tmh an toan cung nhU tac dung khang ung thU. Bai bao nay chimg tdt thdng bao ket qua danh gia tac dung kfiang mdt sd ddng te bao ung thU cua altretamin tdng hdp dUdc.
II. £)6l TU'ONG VA PHU'OfNG PHAP NGHIEN CUU 2 . 1 . Odi tUdng, vat lieu nghien ciru 2.1.1. Chd pham nghien cuU: Altretamin do Trung tam Nghien ciru dng dung san xuat thude Hpc vien Quan y tdng hdp dat tieu chuan DUdc dien My 34.
2.1.2. Vai lieu nghien cdu: Cae ddng te bao ung thU T-47D, A549, Ovcar-3 dUde cung cap bdi Nexeelom Bioselenee, dddc nudi cay va thU nghiem tai Khoa Sinh ly benh - Hpe vien Quan y.
2.2. Phu'dng phap nghien cu'u: PhUdng phap nghien cUu danh gta tae dung khang te bao ung thU vu, ung t h u buong trirng, ung t h u phdi ngUdi in vitro ciia altretamin.
2.2.1. Ky thuit nudi cay tetiio ung thd ngddl Cae ddng te bao ung thUT-47D, A549, Ovear- 3 dUde nudi cay tren dTa nudi cay te bao dUdng kinh i o cm bang mdi trudng nudi cay tUdng il'ng la RPMI, ed bd sung 10% FBS va 1 % Penieiltin va Streptomycin. Sau moi 72 gid, thay mdi trUdng nudi cay eho den khi te bao mpc du sd iUdng (~
80% dien tieh be mat dTa) thi cay chuyen te bao vao cac dTa petri hoac vao dTa 96 gieng de thUc hien cae thU nghiem Trypan Blue va I^TT.
2.2.2. Thiet ke nghien cdu: PhUdng phap nghien eUu danh gia tac dung Ue che sU phat trien ciia te bao ung thU cua altretamin toan phan tren cae ddng te bao T-47D, A549, Ovarc bang thU nghiem Trypan Biue. Danh gia tae dung khang cac ddng te bao ung thu T-47D, A549, Ovcar-3 ciia altretamine bang thU nghiem MTT.
2.2.3. Danh gia tie dung gay dut gay DNA te bao ung thd: Cac te bao dUde ehia thanh eac gieng u vdi altretamin 10|j/ml va dUdc danh gia sau 48h il. Tach chiet va tinh sach DNA tdng sd theo qui trinh hUdng dan eiia nha san xuat (kit QIA Amp DNA^Mini Kit, QIAGEN, Cong hda Uen bang OUe). Mau te bao sau khi nghien ky, hda tan trong 200 ui PBS. Tiep tue tach DNA theo qui trinh tdm tat nhu sau:
1) Cho 20ni Protease K vao dng eppendorf l,5mi. Sau dd cho 200|il dung dich nghien m l u .
2) Cho 200^1 dem AL vao mau sau do lac trdn trong 15 giay.
3) U d nhiet dp 56''C trong l-3h, ly tam ngan.
4) Cho 200(ii eon tuyet dot vao mau, lac 15 giay, ly tam 15 giay.
5) Chuyen toan bd sang cdt lpc va dUng trong dng di kem. Ly tam 8000vdng/pliut trong 1 phiit, bd djch lpc va gltT lat edt.
6J Cho 500^1 dem A W l de rUa cdt. Ly tam 8000 vdng/phiit trong 1 phut sau do bd djeh lpc va giu" lai edt.
7) Cho SOO^it dem AW2. Ly tam 14000 vdng/
phut trong 3 phut sau dd bd djeh tpc, giir lai edt Tiep tue ly tam 14000 vdng/phut trong 1 phut,
8) Chuyen edt sang mdt dng eppendorf 1.5 ml vd triing. Sau do them 260|il dem AE hoae nUdc cat khU ion. U nhiet dp phdng trong 1 phut roi ly
VIETHAM MEDICAL JOURNAL N°2 - APRIL - 2018 tam 8000 vdng/phut tnDng 1 phut.
9) Thu DNA trong ong va bao quan d - 20''C.
Do ndng dp DNA.
Dien di san pham DNA tren Agarose gel 1,5%;
III. KET QUA VA BAN LUAN
3 . 1 . Tac dung khang t e bao ung thu" vu ngu'di cua nguyen lieu Altretamin td'ng htfp tai Viet Nam
3.i.l. Ket qua thiy nghiemJrypan Blue cua nguyen lieu Altretamin iong hdp iren dong tebao ung thd vu
Bang 3.1. Kei qua thd nghiem Trypan Blue trin ddng tebao T-47D
Altretamin 1 pg/m Altretamin 0,5
ill
ug/ml (5)
87,95± 12,52 94,73± 9,08 J) > 0,05
P M > 0,0!
Ps-o > 0,01
Nhom Chu'ng Doxorubicin
10 yg/ml Altretamine
10 lici/ml Altretamine
5 pg/ml Altretamine
1 ^lg/ml Altretamine
0,5 ijg/ml So t e bao
chet 25 27 32 29 25 29
Tong so te bao 398 387 401 412 407 385
P
p >0,05
Ket qua thU nghiem Trypan Blue cho thay nguyen lieu Altretamin tdng hdp d cac ndng dp nghien ciTu khac nhau va Doxorubicin 10 ^glvn\
khdng gay ddc trUe tiep ddi vdi te bao T-47D.
Bieu hien, ty le te bao ehet va tdng sd te bao giu'a cae gieng nudi cay sU dung dieh nguyen lieu Altretamin tdng hdp d eac nong dp khae nhau va Doxorubicin lOpg/ml khdng cd sU khac biet cd y nghTa thdng ke so vdt chUng (p > 0,05).
3.1.2. Ket qua thd nghiem MTT cua nguyen lieu Altretamine td'ng hdp tren ddng te't/ao ung ihd vii
Ket qua nghien cUu tae dung khang te bao ung thU vu ngUdi ddng T-47D ciia nguyen lieu Altretamin tdng hdp dUde xae djnh d eac thdi diem sau 24 gid, 48 gid va 72 gid.
Bang 3.2. Kei qua thd nghiem MTT iren ddng tebao T-47p thdi diem 24 gid
Nhom Chulig (0) Doxorubicin 10
MQ/ml (1) Altretamin 10
liq/ml(2) Altretamine 5
pg/ml t3)
Ty le t e bao song (Vo)
100 84,47± 11,95 87,23± 11,65 86,89± 17,22
P
Pi-o < 0,0!
P2 0 > 0,01 P M > 0,0!
Quan sat dUdi kinh hien vi cho thay thdi diem sau 24 gid: nhu'ng gieng te bao trong mdi trUdng ed nguyen lieu Altretamin tong hdp vdi cac nong dp khac nhau ed sd lUdng te bao ft hdn so vdi cae gieng te bao khdng dimg thuoc.
Thu" nghiem MTT cho thay nguyen lieu Altretamin d ndng dp cang eao thi sd IUdng te bao ung thU sdng sdt cang giam (mat dp quang chat mau giam).
Lieu Ue ehe 50% te bao (IC50) ciia nguygn lieu Altretamin tdng hdp tren ddng te bao ung thUT-47D sau 24 gid la: IC50 = 75,50 [jg/ml.
Bang 3.3. Ket qua thiy nghiem MTT tren ddng tebao T-47D thdi diem 48 aid
Nhdm Chu'ng (0) Doxorubicin 10
ug/ml (1) Altretamin 10 ^lg/mi
121 Altretamin 5 pg/ml Altretamin 1 [jg/ml Altretamin 0,5
Ty le t e b^o sdng (°/o)
71,799± 4,603 85,15± 8,34 85,66± 10,65 99,68± 11,06
I8,26± 15,48 p5.o>0,05 Pi-o < 0,05 p2.o > 0,05 P3-0 > 0,05 P4-0 > 0,05
Tat thdi diem 48 gtd, cae gieng ed sU dun Doxorubicin lOpg/mi ty le cac te bao sdng so vdi Id ehirng giam manh, sir khac biet ed y nghia thdng ke vdi p<0,d5.
Cae gieng ed sd dung nguyen lieu Altretamin tong hdp vdi cac ndng dp lOug/ml, 5pg/ml va Ipg/ml quan sat dudt kinh hien vt cho thay so IUdng te baogiam di rd ret, tuy nhien so vdi nhdm chimg sU khae biet chua cd y nghia thong kevdi p>0,05.
Lieu Uc ehe 50% te bao (IC50) ciia nguyen lieu Altretamin tdng hdp tren ddng te bao ung t h u T-47D sau 48 gid 1^: IC50 = 30,53Mg/ml.
*Tac dung khang te bao ung thU vu ngUcfi sau 72 gid:
DUdi kinh hien vi quang hpc cho thay theft diem 72 gtd sau diing nguyen lieu Altretamin tdng hdp va Doxorubicin lOpg/ml te bao bj ut che phat trien rd ret. Nhat la nhulng te bao moi trudng cd nguyen lieu Altretamin nong dp lOtjg/ml, 5pg/ml. Hinh anh thUa thdt eac te bao bam vao day gieng.
TAP CHt Y HOC VIET NAM TAP 465 • T H A N G 4 - SO 2 - 2018
Bang 3.4. Kei qua thu" nghiem MTT tren dong tdbio T-47D thdi diem 72 g/d
Nhom Chij'ng (0) Doxorubicin 10
pg/ml f l ) Altretamin 10
pq/ml (2) Altretamin 5
pg/ml (3) Altretamin 1
pq/ml (4) Altretamin 0,5
pg/ml (5) P _.
Ty Ie t e bao song ( % )
100 40,553 ± 4 , 5 6 0 81,195± 13,518 82,346± 9,461 85,581± 13,875 85,937± 20,306
p > 0,05 P
Pi-o < 0,05 P M < 0,05 P M < 0,05 p « , > 0,05 Ps-o > 0,05
Tat thdt diem 72 gid, cae gieng ed sU dung Doxorubicin lOpg/ml ty le eac te bao sdng so vdi Id chiTng giam manh, sU khac biet cd y nghTa thdng ke vdi p<0,d5.
Cac gieng cd sU dung nguyen lieu Altretamin tdng hdp vdt d cac nong dp lOpg/ml, 5pg/ml, Ipg/ml va 0,5|jg/ml quan sat dUdt kmh hien vi eho thay sd IUdng te bao giam di ro ret, tuy nhien vdt nong dp lOpg/ml, 5pg/ml sU khae blet ed y nghTa thdng ke vdt p<0,05.
Lieu Uc ehe 50% te bao (IC50) ciia nguyen lieu Altretamin tdng hdp tren ddng te bao ung thUT-47D sau 72 gid la: IC50 = 64,43yg/ml.
3.2. Tac dung khang t e bao ung thu* bieu md ph€ nang A549 ciia nguyen lieu Altretamin tong hcfp tai Viet Nam
3.2.1. Kit qua Hid nghiem trypan Blue cua nguyen lieu Altretamin td'ng hdp tren ddng tdb^o ung tiiUbieu md phe nang AS49
Tten hanh thU tae dung khang ddng te bao ung thU bieu md phe nang A549 ciia altretamin tdng hdp, ket qua dddc trinh bay d bang 3.93.
Bang 3.5. Ket qua thvT nghiem Trypan Blue
nghien cirtj khac nhau va Doxorubicin 10 pg/ml khdng gay ddc trUc tiep dot vdi te bao A549.
Bleu hien, ty le te bao chet va tong sd te bao giu'a cac gieng nudi cay sU dung dich nguyen lieu Altretamin tdng hdp d eac nong dp khac nhau va Doxorubicin 10 pg/ml khdng cd sU khac biet cd y nghta thdng ke so vdi chimg (p > 0,05).
3.2.2. Kei qua ihd nghiem MTT aia nguyen lieu Altretamin td'ng hdp tren ddng tebao ung thUbieu md phe nang A549
Ket qua nghien cUu tac dyng khang te bao ung thU bieu md phe nang ngudi ciia nguyen lieu Altretamin tdng hdp dddc xae djnh d cae tiidi diem sau 24 gid, 48 gid va 72 gid nhU sau:
Bang 3.6. Kei qua thd nghiem MTT bdn ddng te bio A549 thdi die/n 24 gid
Nhom Chu'ng Doxorubicin 10
pg/ml Altretamin 10
pg/ml Altretamin 5
pg/ml Altretamin 1
pq/ml Altretamin 0,5
pg/ml S S t e bao chet
20 26 27 24 21 27
Tong so t e bao
385 376 400 395 388 385
P
p >0,05 STT
1 2 3 4 5 6
Nhom Chu'ng (0) Doxorubicin 10
pq/ml (1) Altretamin 10
pq/ml (2) Altretamin 5
pg/ml (3) . Altretamin 1
pq/ml (4) Altretamin 0,5
ug/ml (5) P
Ty Ie t e bao song ( % )
100 61,22± 26,12 80,31± 20,11 89,97± 22,48 93,29± 19,72 90,38± 22,56 P M < 0 , 0 5
P Pl-0<
-
0,05 P2-0<
0,05 P M >
0,05 P4 0>
0,05 Ps-o>
0,05 Quan sat dudi Idnh hien vi cho thay thdi diem sau 48 gid: cac gieng \& bao cd sU dyng Doxorubicin 10^Jg/ml, t / ie te bao sdng thap nliat, sU khac biet cd y nghTa thdng ke vdt p<0,05.
Nhihig gteng te bao trong mdt trUdng ro nguyen lieu Altretamin tdng hdp vdi ndng dp lOpg/ml, sU khac biet ed y nghTa thdng ke vdt p<d,05.
Lieu Uc ehe 50% te bao (IC50) ciia nguyen lieu Altretamin tong hdp tren ddng te bao ung t h u A549 sau 24 gid la: IC50 = 36,82pg/ml.
Bang 3.7: Kei qua thU nghiem MTT trdn ddng tebao A549 thdi diem 48 g/d
Ket qua thU nghiem Trypan Blue cho thay nguyen lieu Altretamin tdng hdp d cac nong do
Nhom Chu'ng (0) Doxorubicin 10
pg/ml (1) Altretamin 10
pg/ml (2) Altretamin 5
pq/ml (3) Altretamin 1
pg/ml (4) Altretamin 0,5
pg/ml (5) P
Ty Ie t e bao soiig ( % )
100 49,31± 3,24 67,35 ± 6,99 73,86 ± 11,00 78,43 ± 7,56 76,10 ± 14,23 P2-1 <0,05
P Pl-0<
0,05 P2-0<
0,05 P3-0<
0,05 P4<1<
0,05 Ps-ii<
0,05
VIETMAM MEDICAL JOURNAL N°2 - APRIL - 2018 Tai thdi diem 48 gid, cac gieng cd sU dung
Doxorubicin lOpg/ml va va eac gieng ed sU dungnguyen lieu Altretamin tdng hdp vdi cae ndng dp lOpg/ml, Spg/ml va Ipg/mi, 0,5Mg/ml quan sat dudi kfnh hien vi eho thay sd lUdng te bao giam di rd ret, ket qua do mat dp quang eung cho ket quatUdng ddng, ty le cae te bao sdng d tat ca cac Id nghien ciili giam manh, so vdi nhdm chUng sU khac biet ed y nghTa thdng ke vdi p<0,05.
Lieu Uc che 50% te bao (IC50) eiia nguyen lieu Altretamin tdng hdp tren ddng te bao ung thu A549 sau 48 gid la: IC50 = 26,73pg/mi.
Bang 3.8. Ket qua thd nghiem MTT tren dong tdbSo A549 thdi diem 72 gid
ng/ml (2) Altretamin 5 pg/ml Altretamin 1 |jg/ml Altretamin 0,5
pg/ml (5)
71,20 ± 12,95 P3-0 < 0,OS
76,26 ± 13,47 72,90 ± 10,94 P2-1 <0,05
tVo<0,OS Ps-o < 0,05
Nhdm ( 1 ) ChUng (0) Doxorubicin 10
Mg/ml (1)
% t e bao song s d t ( 2 )
100 25,64 ± 9,26 69,48 ± 11,66
pi-o < 0,05 P2-0 < 0,05
Tai thdi diem 72 gtd, cae gteng cd sU 1 Doxorubicin 10[jg/ml va va cae gieng chii dungnguyen lieu Altretamin tdng hdp v i ^ c &
ndng dp lOpg/ml, Sp^/ml va Ipg/mi, 0,5Mg/nil quan sat dUdt kfnh hien vi cho thay sd lUdng te bao giam di rd ret, ket qua do mat dp quang cung cho ket qua tUdng ddng, ty 1$ cac te bao sdng d tat ca cac Id nghien cuU gtam manh, so vdi nhdm chUng sU khac biet cd y nghla thdng ke vdi p<0,05.
Lieu Uc che 5 0 % te bao (IC50) ciia nguyen lieu Altretamin tong hdp tren ddng te bao ung thUA549 sau 72 gid la: IC50 = 41,18|jg/ml.
*Tac dung khang tebao ung ff"{bieu/nd phe nang A549 ngddi sau 72 gid:
Hinh 3.1. Tebao ung thUAS49 sau 72 gidnudi cay dmdi trUdng khdng bdsung miu nghien cdu (A) v^ bdsung altretamin 10 pg/ml (Dg phdng dai 400X).
3.3. Tac dung khang t e bao ung thu' j Altretamin 0,5 I buong tru'ng ngu'di cua nguyen lieu
Altretamin t d n g hcfp t a i Viet Nam 3.3.1. Ket qua thd nghiem trypan Blue cua nguyen lieu Altretamin idng hdp tren ddng tebao ung thu budng tru'ng Ovcar-3
Tten hanh thU tac dung khang ddng te bao ung thu budng trimg bvcar-3 ciia altretamin tdng hdp, ket qua dddc trinh bay d bang
Bang 3.9: Kei qua thd nghiem Trypan Blue tren ddng ti'bao ung thU buong tru'ng Ovcar-3
42 I 385 T
Nhom Chiitiq Doxorubicin 10
pq/ml Altretamin 10
pq/ml Altretamin 5
pq/ml Altretamin 1
pq/ml Sote bao chet
39 41 37 36 38
Tong so t^bao
412 423 398 400 389
P
p >0,05
Ket qua thir nghiem Trypan Blue cho thay nguyen lieu Altretamin tong hop d cac nong do nghien cull khac nhau va Doxorubicin 10 pg/ml Ichong gay (Jpc trtfc tiep doi vdi te bao Ovarc - 3._Bieu hien, ty le te bao chet va tong so te bao giij^ cac gieng nuoi cay sit dung dich nguyen lieu Altretamin tong hdp 6 cac nong do l<hac nhau va Doxorubicin 10 pg/ml khong co sir khac bift CO <i nghta thong ke so vdi chiJng (p > 0,05).
3.3.2. Oanh gia tac dung gay diit gay DNA t e bao ung t h y cua Altretamin
Tien hanh danh gia tac dung gay dut gay DNA te bao ung t h u vdi cac ddng te bao T-47D, A549, Ovarc, ket qua dUdc trinh bay trong Asns 3.10^3 hinh 3,2.
Bang 3.10. Ndng do trung binh DNA tai cac gieng nudi
Mau thu' Chu'nq
T e b a o T - 4 7 D
Niong dd DNA (ug/til) ^
35,1±9,6 T e b a o
A549 40,7±12,5
T e b Ovare' 37,7±13i!
TAP CHt Y HOC VIET HAM TAP 465 - THANG 4 - SO 2 - 2018 Altretamin
lOnq/ml 29,1±6,2
>0,05 33,9±9,5
>0,0S
31,5±10,2
>0,05 UI vu LTTPhoi LTTBuong Chu'ng- Chu'ng- tru'ng
™retamin Altretamin Chifng-
^ Q ^ r ~ i Aisa™,
• • •
Hinh 3.2. Ket qua dien d/DNA Ket qua cho thay, tat eae gieng nudi cac ddng te bao Ichac nhau, mac dii nong dp DNA eiia cac gieng chirtig cd xu hUdng eao hdn, nhUng dmh lUdng DNA trong eac gieng nudi khdng cd sU khac blet cd y nghla thdng ke giu^ gieng chUng va gieng cd Altretamin.
IV. BAN LUAN
Ket qua nghien culJ ciia chiing tdt cho thay budc dau Altretamin da cd tae dung Uc che sU phat trien, ddng thdi ed tac dung lam dirt gay cac DNA ciia te bao ung thU tuyen vu, ung thU bieu md phe nang va ung thU budng trimg. Tae dyng ciia Altretamin ddi vdi ung thU buong trUng da dUdc nhieu tac gia chUng minh tren thuc nghiem va tren ngUdi chii yeu giai doan ttiU nghiem pha I I tren ngUdt. Keldsen N (2003) nghien CUIJ tac dyng Altretamin trong dieu trj ung t h u buong trimg da dieu tri tan ^ u nhUng tat phat trong vdng 6 thang, cae benh nhan dUde dieu trj Altretamin vdi lieu 260mg/mVngay dieu tri 4 lan trong hat tuan va lap lat sau 4 tuan, ket qua danh gia sau 2 lan dteu trj 26/31 benh nhan dap img vdi dieu tri, thdi gian sdng trung binh tien trien la 10 tuan, cd rat ddc tinh ve huyet hpe, chil yeu ddc tfnh vdi dUdng tieu hda, tae gia ket luan khdng nen chpn dieu tri Altretamin eho benh nhan ung t h u buong trUng tai phat cd di can hach bach huyet, tuy nhien nen chpn Altretamin cho nhutig benh nhan Uu tien dieu tri bang dUdng udng va ndi ed dieu kien kinh te chu^ phat trien.
Tuy nhien Altretamin dUdng nhU chu^ cd tac dung nhteu ddi vdi cac ung thU khac ngoai ung t h u buong triing. NhU vay, tren thUc nghiem eho thay ket qua nghien eUu ciia chiing tdi cho bUde
dau ehirng minh Altretamin da ed tac dung Uc ehe sU phat trien, ddng thdi ed tac dung lam diit gay eae DNA ciia te bao ung thU tuyen vii, ung thU bieu md phe nang va ung thU budng trUng.
Oieu nay can tien hanh tren thuc nghiem d dpng vat va tien ahnhf tren ngUdi d Viet Nam, rat cd the trong thdi gian khdng xa altretamin diidc sir dung eho nhu'ng benh nhan ung thU vdt ldi the diing dUdng udng, dieu trj ngoai tru va eho nhij'ng ngudi cd dieu kien kinh te khd khan.
V. KET LUJSiN
- ThU nghiem Trypan Blue, ty le te bao chet va tdng sd te bao giO'a cac gieng nuoi cay sU dung djch nguyen lieu Altretamin tdng hdp d cae ndng do 10 pg/ml, 5|jg/ml va 1 pg/m'l, 0,5 pg/ml va Doxorubicin lOpg/ml khdng ed sU khae bi|t (p>0,05) dot vdi te bao ung thU vu, ung thU bieu md phe nang A549, te bao ung thU budng trUng Ovear-3.
• ThU nghiem MTT cho thay nguyen lieu Altretamin tong hdp Ue che sU phat trien eiia te bao ung thU vu giam d thdi diem 72 gid vdi ndng dd^ lOpg/ml, 5[jg/ml (p<0,05); te bao ung thU bleu md phe nang ddng A549 d thdt diem 24 gid vdi ndng dp lOpg/ml, thdi diem 48 gid va 72 gid vdi eae nong do lOpg/ml, 5pg/ml va Ipg/mi, 0,5pg/ml (p<0,05).
- Tae dung gay dirt gay DNA te bao ung thU vdi cac ddng vdi eac ddng te bao T-47D, A549, Ovarc ciia Altretamin khdng cd sU khac biet vdi nhdm eh Ung.
TAI LIEU T H A M KHAO
1. Phan Dinh C h i u , Vu Binh DUdng, Nguyen Thi Thu H i n g , Bill Thj Thu Ha ,(2014J, Nghien euU quy trinh san xuat altretamin la thuoc dieu tri ung thu. Tm ehi dUcJc hoc so 462; trang 2-6.
i. Nguyen Ba Du'c (2003);H6a chat dieu tn ung thU, NXB Y hoc HN,tr. 130-137.
3. C. Rhoda L ^ , Diana Faulds (1995), "Altretamine A Review of its Pharmacodynamic and Pharmacokinetic Properties, and Therapeutic Potential in Cancer Chemotherapy", Drugs June 1995, Volume 49, Issue 6, pp 932-953, 4. Fabian C J . , Rasmussen S , Stephens R, Haut
A , Smith F, BalcxRak S , Tranum 8, (1979), "Phase II evaluation of hexamethylmelamine in advanced breast cancer: a Southwest Oncology Group study". Cancer Treatment Reports [01 Aug 1979, 63(8): 1359-1361].
5. Keldsen N l , Havsteen H, Vergote I , Bertelsen K, Jakobsen A, (2003), ".Altretamine (hexamethylmelamine) in the treatment of platinum-resistant ovarian cancer: a phase II study", Gynecol Oneol. 2003 Feb;88(2): 118-22.
o. WHO (2017), "Cancer Statistics - National Cancer Institute",https://www.caneer.gov/about-caneer/
understanding/ statistics.
