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Me

dic
ina
lPl
ant
,201
5,6(9-10):4-7


nhibi
tor
y Ef
fec
ts ofBlac
k Garl
ic Ex
trac
ting Sol
uti
on on t
he
Gro
wthofMiceColonCancerHt
29

Chunf
engYU,Ai
huaHOU,Yue
junMU,We
iLI
U,Lai
f U
uLI
Onc
olo
gyo
fYa
nta
iHo
spi
ta
lofTr
adi
ti
ona
lChi
nes
eMe
dic
ine
,Ya
nta
i,S
hando
ng2
640
00,Chi
na

Abst
ract [Obj e
cti
v e
]Tor es
earc
hthei nhi
bit
oryeffe
c t
sofblackgarl
icextr
acti
ngsolut
iononthegro
wtho fcol
oncancerHt29thr
oughPI3K/
AKTs i
gnalpathway.[Me t
hod]Th etumorbea
ringC5 7BL/6mo usemo delwasesta
bli
shed.Micewe r
egivend i
ff
eren
tc on
centr
ati
onsofbla
ck
ga
rli
cextrac
ti
n gsol
uti
on sandPI3Kinhi
b i
torLY294 0
02.Att hee ndofthetes,t
t hetumorswerepeele
do f
fa n
dwe ig
hedtoc al
cul
atetheinhi
bi

oryrat
eo ftumorgrowth.Ak tpro
tei
ne xpr
essi
onwa sd ete
ctedb yimmunoh i
st
ochemis
try.Badan dPTENp r
otei
ne xpr
essi
onwa sanal
yzedby
West
ernb l
ot.An dthee xpr
essi
onofCas p
ase9wasd etect
edb yRealt
imePCR.[Re s
ults]Thetumorinhi
b i
tor
yrateofblackgar
li
ce xt
rac
ti
ng
so
lut
iononmi cecolonc anc
erreac
hed4 9.93%.Bl a
ckg arl
icextr
acti
ngsoluti
onrest
ric
tedtheexpre
ssi
ono fAktandp Bad,b utenhanc
edthe
ex
press
ionofPTEN,Ba dandc as
pase
9.[Co nclusi
ons ]Blackg ar
licextra
cti
ngs ol
uti
onmi g
h tre
str
icttheg r
owtho fcol
onc anc
erHt 
29

hroug
hPI 3K/ AKTs i
gn alpa
thway.
Keywo rds Bl ac
kg arli
ce x
tra
cti
ngsol
u t
ion;Ht29;PI 3K/Ak tsi
gnalpathway

1 I
ntr
oduc
tio
n fr
om Ame rsc o; BCA wo r
kings o
luti
on, Ak t, Ba d, PTEN,
Atp res
e nt
,t r
eatmentsofc ol
onc a ncerarema inl
yo peratio
na nd Caspase9a ndβ a
cti
na ntibody,h orser
adishp erox
ida s
el abe l
ed
chemo t
h er
apy.Ho wever,chemo t
h era
p ydrugh aslet
hali
tyt ob ot
h goatan t
imu se ,h ors
erad i
shp er
oxidaselab el
edg oata nti
rabbit


u morc ell
sandn ormalcell
s.Att h esamet i
me ,ithassidee ff
ect
s andECLPl usl uminescenc ek i
twe rea l
lp urchasedf romS i
gma ;
onl i
ver ,k i
dney,ma r
row,u r
inarys ystema ndg ast
roin t
est
ina
l DMS Owa sp urchasedfromAn huiAn t
eBi ochemi st
ryCo .,Lt d.;

ract.Th eref
ore,itisurgenttofi
n dingane f
fect
iveandl owtox
ici
 DEPCwa sp urch a
sedf romS igma;RNAi soPl u s,r evers
et ran

ya ntit
u mordrug.Re sear
chesh avefo u
ndo utthatgar
licp romot
es scr
ipti
onk ita ndRe alt
ime PCRk i
twe reallp ur
c hasedf r
omTa Ka

h etumo rc e
llapoptos
is,c hangest umorc el
lc y
cle,p rot
e ct
sthe Ra;LY2 94 00 2wa spurch asedfromPr omegaCo rp or
ati
o n;an do t
h
organi
smf romr a
diati
ona ndsoo n[1].Ph os
phoinos
it
ide3 kina
se/ erconve nt
io nalr eag
entswe rep urchas
edf r
omBe y ot
imeI nsti
tuteof
seri
ne/threoni
nep rot
einkinase(PI 3K/ Ak t
)i sanimp ortantsi
g Biote
chn ology .
nalme diat
orinc el
ls,wh i
chp ar
ticipate
sint heproce
sso fp r
oli
fer
 2.1.3  Ex perimentala nimala ndc el
ll i
ne .A t o
talo f40ma l

ati
on,a poptos
isandmi gra
tionofma n
yt umorcell
s.Du etot heim C57BL/ 6mi c ea t6-8we ekswe rep urchasedfr om Ex per
imen ta

port
ancet ot heoccurre
n c
e,d e
ve l
o p
me nta ndrecover
yo ftumor Anima lCe n teri nPe k i
ngUn i
onMe dicalCo llege.Th eirb ody
dise
ases,PI 3K/Ak th a
sb ecomet her esea
rchh ot
spoti nr ecen
t weightwa sb et
we en1 8a nd2 1g .Mi cewe ref e
du nderS PFg rade
[2]
years .Ba sedonthese,blackg arli
cwa sprepar
edintoe xtr
acti
ng fe
ed i
ngc o
n dition.Mi cec olonc ar
cinomaHt 
29c el
lswe rep r
ov i
d
solu
tionsinthisresea
rch.Th eeffecti
vec omponent
swe ree xt
ract
ed edb yth eEx perimentalCe nterinBi ngz
houMe dicalCo l
leg
e .

o rther es
earchont hetranspl
antationtumo rofcolonc ancerHt 2.2 Me tho ds
29.I nhibit
oryeffec
tso fblackg a r
li
ce xtract
ingsolut
iono nthe 2.2.1 An ima lmo deli
n ga ndt es
tg r
ouping.Ht 29c ell
sa tloga

u moro fcolo
nc ancerwereo bs
erved,a ndi t
srela
tio
nt oPI 3K/Akt ri
thmicp h
a sewa swa sheda ndd i
g e
stedb y0.2 5% t rypsn,a
i nd
si
g n
alp athwaywa sdis
cussed. th
e ncen tr
ifug eda t10 00r /mi nfor5mi n.Ce lls u
rvivalratewa s
detec
tedb y0. 4% t r
yp anb lues ol
uti
on(s u rvi
va lrat
e> 9 0%).
2 Ma
ter
ial
sandme
tho
ds Thec ellco ncen tr
ati
onwa sadjustedto1×1 06 c
ells/mL.S kind is

2.1 Ma t
erial
s in
fecti
onwa sc arr
iedou tb y75% a lcohol
.0. 2mLHt 2 9cellsu s

2.1.1  Pl an
tma ter
ial
s.Bl a
ckg ar
li
cwa sp r
ovid
edb yDa l
ian pensi
onwa si n ocula
tedt ot heb acko feachmo u se
.Wh ent umor
Hu ag
uGa rl
icIndustr
yCo .,Ltd.Ga r
li
cwa sgri
nded,soake
di n diameterfo rs ubcutaneousi nocul
atio
nwa s8-1 0mm,mi cewe r

95% e t
hanol
,f i
lt
rat
edb yvacuump u
mp,c oncen
trat
edandster
i ra
n domlyd i
v i
d edintofou rgroups,wh ichwe reb l
a n
kc o nt
rolgroup
li
zed.Th u
s,t heblackgarl
icextr
act
ingsol
uti
on(a bou
t1g/ mL (intr
aperit
o neali nject
iono f0 .2mL o f0 .9% Na Clf ore ach
crudedrug)wa sobt
ainedandstor
edat4℃ f orfut
ureuse
. mou s
e),b lackg arl
iclow concent
rat
iong r
ou p(i n t
raperi
tonealin
2.1.2 Ma i
nr eag
ents.Feta
lc a
lfser
uma ndRPMI 1640cult
ure je
ctio
no f0. 2mLo f50mg/ mLb lac
kg arl
ice xtr
a ct
ings olu
tionf o

med i
umwe rebothpurcha
sedfr
omHy clo
ne;try
psinwaspurc
hased eachmo u s
e ),b l
ackg a
rlich i
ghconcent
rati
ong roup(i ntrape
rit
on ea

in
jecti
ono f0 .2mLo f1 00mg/ mLb lac
kg arli
ce xtract
ings ol
uti
o n)
Re
cei
ved:Augus
t2,2015  Accept
ed:Sep
tember23,201
5 andLYg r
ou p(i ntrap
eritonealinje
cti
ono f0. 2mLo f25u g/mLLY
Cor
resp
ondi
ngaut
hor
.Emai
l:y
gq19821
009@1 63.co
m 294002).I n j
e cti
onwa sc arri
edo utforinall1 4d ays.
Chunf
engYUe
tal
.Inh
ibi
to
ryEf
fe
ctso
fBl
ackGa
rli
cEx
tra
cti
ngSo
lut
io
nont
heGr
owt
hofMi
ceCo
lonCa
nce
rHt
29 5

2.2.2 Ca l
culationo fthei nhibitionr ateo ftumo rg rowth.Wh en ed.Af terwellmi xe
d,t hes olutionwa splace
df o r3mi na ndthen
thete s
te nded,mi cewe rek ill
e d,a n dt heirb odys urfacet umor centr
ifuged.Th us,wh i
tet abletwa sobtai
n e
d.Af terdrie
d,4 0μL
wa stakeno f
fa ndwe i
g hed.Ac cordingt ot heeq uati
ono fin hibiti
on enzymewa sadd e
df ord is
soluti
o n.RNAp uri
tya ndc ont
entwe r

rateoftumo rg rowth=(a ve raget umo rwe ightinc ontrolg roup- detect
edb yu lt
ravio
letsp ect
roph ot
ome t
er.RNAp uri
tywa sdetec

averagetumo rwe i
ghti nt r
eatme ntg roup)/a ve r
ag etumo rwe ig
ht tedb ynu cl
eica c
ids pectr
ome ter.OD260/ OD280 waswi thi
n1. 8-
inc ont
rolg r
oup×1 00%.Th ei nhibiti
o nr at
eo ftumo rgro wthwa s 2.2,i ndicat
ingg oodRNA p ur i
ty.Pr i
me rd esigna nds ynt
hesi

calculat
edi ne acht estg r
oup. werea sfoll
ows:1μgs amp l
ewa sr e
versel
yt r
an scri
bedt ocDNA.
2.2.3 Ef fect
so fb lackg arli
ce x t
ractings oluti
o no nth eAk tp r
o Primerwa ss ynt
he s
ize
db yTa k araBiotechnol
og y(Da l
ian)Co .,
tei
ne xpr
essiono fHt 29t r
a nsplantedt umo rtissues.Ak tmo no Ltd.Th eu pstr
eamp rimerwa sc c
ttgt
gt
c c
tactccacctt
c,a ndthe
clonala nt
ibodywa ss el
ected;i mmu no hist
oc hemicals t
ain i
n gwa s down s
treamp rimerwa sa t
ctgcttg t
aagtcc
c t
tt
cg.Re act
ions ys
tem
carri
edo utb yS ABC me t
h od:mi c
et u morwa sf ixedb y4% wasp reparedaccordi
ngt ot hefo l
lowingcomp onen t
s:① Re agent

parafor
ma ldehydef ixedf l
uidf or6h.Af terwa she db yf l
o wwa t
er SYBR Pr emixExTa q(2×)(1 0μL);PCRf orwardp ri
me r
(10
for6h,d ehyd r
a t
iono fgrad i
en te t
ha no lwa sc arri
edo ut.Af t
er μmo l)(0.4μL);PCR r eversep r
imer(1 0μmo l
)(0.4μL);
tr
a ns
p ar
enc ean dwa x i
n g,4μm s l
icewa sp repared.Af t
e rd ewa DNAt emp l
ate(2. 0μL),d H2O(7. 2μL),at otalof2 0.0μL
xing,r et
ri
e vi
nga nti
g en,c oolingt or oo mt emp eratur
ea nds erum solut
ionwa swe l
lmi x
eda n damp l
ifi
edb yRo t
orgen e3000Re al
ti
me 
blocking,1∶2 00Ak tmo noclona la ntibodya nds econda ntibody PCRa mp l
if
ier;thereac t
ionp rogramo fRe a
lTi mePCR:9 5℃ 3 0
we r
ea d ded.Ba sedo nS ABC s t
aining ,c o l
ord eve
lopin ga g e
nt S;PCRr eacti
ona t95℃ 5S 
,6 0℃ 2 0S ,4 0c ycles
;me l
ti
ng
AECwa sa dded.Mi ldr edyeingo fh aema toxyli
nwa sc on ducted. curvewa sa na
lyzed.
Afterd ehydrati
ona ndmo un t
ing ,t hei mmu noh i
stochemic als li
ce 2.3 St a ti
st
icaltreatme nt S PSS13.0s of
twarewa susedford a


wa sobservedu nd erh ighp o
we rl ens(×4 0)o fOLYMPUSBX5 1 taa n
a l
yses.Co mparis
ono fs amp lerat
ewa sc arri
e doutb yχ t
est

(FN2 2).Fi vev is
ua lfi
e l
dswe res el
e ctedf r
ome achs li
ce .Th er
e An dc o
mp ari
sono fsamp leme anv aluewa scon ductedb yv a
rian
ce
we r
ei na l
lt h
re er e
p eti
ti
o ns
. analysi
s.
2.2.4 Ef fect
so fb lackg arli
ce xtr
actings olutiono nth eBa da nd
PTENe xpressioni nt ranspl
a ntedt umo rt iss
u esb yWe sternBl o
t. 3 Re
sul
tsa
nda
nal
yse

Asf o reac hmo use,a bout3 0mgt umo rtissueswe rec u ti nt
o 3.1 I nhibit
orye ff
e ct
so fblackg arl
ice xt
racti
ngsoluti
ono n
pieces,a ndtran sf
erredt o20 0μLc ellly s
iss ol
u t
ion.Th es oluti
on theg rowtho ftransplantedtumo ro fmi cewithcoloncancer
wa scentri
fugeda t120 00r /minf or1 0mi nu nde r4℃.Pr ote
in Ht 
29 Mi c
ewe reprocessedbyd r
ugsf or14d,a ndthenk i
ll
ed.
concentrat
iono fs upern a
tantwa sd et
ec t
e db yBCAme t
h od.5 0μg Thec omplet
et u
mo rb od
ywa sc ol
le
cted;a ndtheinhi
bit
oryra
teof
protei
nwa su se dfors amp l
el oad ing.Pr o t
einwa si sol
atedb y1 5% tumorgrowthwa scalcul
a t
ed.Ta bl
e1r epor
tedtheres
ult
s.Co m
SDS PAGEg ele l
ectrophoresis.PVDFme mbran eo fp rotei
nwa s paredwithb l
ankc ontrolgrou
p,t het umorg r
owthinlowdos
age
tr
a ns
ferredb y7% s kimmi l
ke nc l
osedme thodfor1h,s o akedi n groupo fblackg ar
li
ce xt
ract
ingsoluti
on,h igh
dosa
geg rou
po f
1×TBS Tb uf
fer,a nds l
owlywa sh e
do ns haki
n gt abl
ef or5mi n. blackg ar
li
ce x
tra
c t
ings ol
uti
o nand LY g r
ouph ads i
gnif
ica
nt
Prima r
ya nti
bo dyo fc orr
espon dingc onc entrat
ionwa sadd ed.i ncu dif
fere
nc e
s(P <0. 05).Co mpare
dwi thp os
iti
vecont
rol(LY)
batio
nwa sc arri
e do uta t4℃ f oro v ernight.Th eso l
u t
ionwa s group,lowdosag
ea ndh i
ghdos
ageg ro
u psshowednos i
gnif
ica
nt
wa sh
e dbyTBS Tf o
rt hreetime swi th5mi nf ore achtime .Un d
er dif
fere
nc s(P >0.
e 05).
Ta
ble1 Ef
fec
tso
fblackg
arli
cext
rac
tingsol
uti
onont
het
rans
pla
nte

roomt emp era
tu r
e ,itwa sincu batedwi t
hd il
uteds econda ntibody

umorgrowt
hofcol
onca
ncerHt2
9(x珋±s)
for1h.Wa shingwa sc arri
edo u tforth reetime sb ythes ameme t
h
Averag
et u
mo r Aver
ageinhib
ito
ry
od.Af terECLPl usc olo
rd evelop ment,me mb ranewa sp u tinECL Gr
oup
wei
ght∥g rat
e∥%
chemi ca
lf luo
resc entreagentf orre a
c t
ionf or3mi n.X p hotograph Blankcont
rolgro
up 7.33±0. 1
2 -
icfil
mwa se xp osedi nd arkb ox .Re se archr esult
swe rea na l
yzed Lowdos
ageg r
oupofbla
ckgarl
icex 5. 45±0. 6
4 25.24#
tr
acti
ngsol
utio

qua n
tit
ativel
yb yGe lProAn alyzer.Av e r
a g
ea bsorbancywa sd e
Highdo
sag
eg rou
pofblac
kgarl
icex 3. 67±0. 3
6 49.93#
tecte
db yQu antit
yOn es oft
wa re. tr
acti
ngsol
utio

2.2. 5 Ca spase9mRNAe x pressiond etect
e db yRe altime PCR. LYg rou
p 3.78±0. 2
7 48.43
Note:n=1 0; ind
ica
teds
tat
is
ti
cald
iffer
encescompa
redwi
thbl
ankc ont
rol
50mgt umo rt i
ssu e
ss t
oreda tl owt emp eratur
ewe reg ri
n dedi nt


rou
p(P <0.
05);#i
ndi
cat
edn
osi
gni
fi
can
tdi
ff
ere
nce
sco
mpa
redwi
thp
osi

powd er.Af t
era dd i
n gliquidn itrogen,t h ep owd e
rwa stran sfe
rred
intoh omog e
na t
ec entri
fuget ubea ndp laceda tr oomt emp era
t u
re t
iv
eco
ntr
olg
rou
p(P >0.
05).

for5mi n.Ce ntrifu


g ati
onwa sc arri
e da t1 20 00g/ minf o r5mi n 3.2 Ex pre
ssi
ono frela
tedg enepr ote
ine xpres
sionofc ol
on
und e
r4℃.Th es upernatantwa st ransferredintoan ewc entri
fuge cancertumortis
suesindiff
erenttestgroups Ex pres
sio
no fco
tube.0. 2mLc hloroformwa sad dedi ntoh omogen i
zedl ysate.Af  lo
nc ancerHt
29tumortiss
uesAk twa sdet
ecte
db yimmu nohis
to
tersh a
kingf or1 5S th esolutionwa sp laceda tr oomt emp era
t u
re chemist
ry;andFig.1ill
ust
rat
edt heresul
ts
.Co mparedwit
hb la
nk
for5mi n.Ce ntrif
ug at
ionwa sca rri
eda t1 20 00g/ minf or5mi n, cont
rolgrou
p,e x
press
ionoftumo rti
ssuesAktr e
duceda f
terpro
thes uper
n at
an twa sc oll
ected.An d0. 2mLi sopropanolwa sa dd cess
edb yLYa ndb l
ackgarl
icextrac
ti
n gsol
uti
on.
6 Me
dic
ina
lPl
ant 2
015

No
te:A.Bla
nkcont
rolg
roup;B.Lo
wdos
ageg
rou
po fbla
ckg a
rli
c;C.Hi
ghd
osa
geg
roupofbl
ackgar
li
c;D.Po
sit
iv
eco
ntr
olg
rou
pLY.
Fi
g.1 Expr
ess
ionofrel
atedge
nepro
tei
nAkto fc
o l
onc a
ncertumort
is
suesi
ndi
ffe
rentt
estgr
oups
3.3 Eff
ect
so fbl
ackg a
rli
ce xtr
act
ingsol
uti
onontheexpres
 Compar
edwithb l
ankcont
rolgro
up,ex
pre
ssi
ono
fBadinb l
ack
si
onofBada ndPTENpr ot
einsinB16cel
lsbyWe st
ernBlot
 ga
rli
cg r
oupsenhanc
eds i
gnif
ic
antl
y;a
ndPTEN ex
pre
ssi
onin
ti
ng Expre
ssi
onofBada ndPTENp rot
ei
nsinmicet
umorti
ssu
es cr
ease
do bv
iou
sly(Fig.2).
wasan
aly
zedbyWe st
ernBlot
ti
n g
.LYwa sposi
ti
veco
ntr
olgr
oup.

Fi
g.2 Ef
fec
tso
fbl
ackg
arl
ice
xtr
act
ings
olut
iono
nthee
xpr
ess
iono
fBa
dandPTENpr
ote
insi
nB1
6ce
ll

3.4  Cha ngeso fCaspase
9mRNA e xpressi
onbyRe a
lti
me
PCR  Fl uoresce
ntq uan
tit
ati
onPCR d etec
ti
onf oundo utthat
Caspas
e9mRNA e xpr
essi
onr e
ducedind if
fer
entg r
oups,c o
m
par
e dwit
hb l
a n
kc ont
rolgro
up.Amo ngthem,h ighd
osagegroup
ofb l
ackg ar
licextr
act
ings ol
uti
onr e
duceds i
gnif
icant
ly(P <
0.05) comparedwithb la
nkc ontr
olg r
oup,s ho
wings tat
is
ti
cal
di
ffer
ence
s.Ho wever,hig
hdosag
eg r
ouphadn osi
g n
ifi
cantdi
ff
er
enceswit
hPI 3KLYp o
sit
ivecont
rolgr
oup(P>0. 05).

4 Co
ncl
usi
onsa
nddi
scus
sio
ns
Prol
if
erat
io
na ndapopt
osi
softumorcel
lswe reinequil
ibr
iumu n
dernormalsi
tua
ti
on,soa stoensur
etheh ealt
hyd ev
elopmentof Fi
g.3 Ef
fec
tso
fblac
kg ar
lice
xtr
act
ings
olut
iono
ntheCa
spa
se
or
ganis
m.On cetheequil
ibr
iumwasdest
royed,tumordis
easesde 9mRNAe x
press
ion
vel
opedandd et
eri
ora
ted.PI3K/Akthadi mp o
rta
ntp hy
siol
ogi
cal l
is
m,t
ran
sla
ti
oni
nit
ia
ti
ona
nds
oon[3-4].PI
3K,c
omp
ose
dofr
eg
fu
ncti
onstotumorcel
ls,whichr
egul
atethep r
oli
fer
ati
on,me t
abo
Chunf
engYUe
tal
.Inh
ibi
to
ryEf
fe
ctso
fBl
ackGa
rli
cEx
tra
cti
ngSo
lut
io
nont
heGr
owt
hofMi
ceCo
lonCa
nce
rHt
29 7

ul
atorysubun i
tp 8 5a ndc at
alyti
cs ubu ni
tp 1 10,wa sad i
me rk i
 showedthatblac
kg arl
icextr
acti
ngsolut
ionhads i
gnif
icantinhibi

nase.Th esti
mu lat
e dPI 3Kg en er
atedt hes econ dme ss
e nger
sPI P3 to
rye ff
ect
so nHt29micetranspl
ante
dt umor.Immu nohis
toc
h em
onp l
asmalemma .PI P3wa stransf
erredt oc y t
o me mb r
an ethrough is
try,We st
ernBlot
tingandRe al
timePCRwe r
eu sedtod et
ectthe
si
gnalregulat
e dk inaseAk twithPHs truct
u re,s oa st oc hangethe expres
sionofprot
einsinvol
vedi ntumortis
sues.Re sult
ss howed
or
iginalconfor
ma t
ion.PDK1i nAk te nvelopec au sedd own s
tream th
a tblac
kg ar
li
ce x
tract
ingsolut
io
nr edu
cedth eexpress
ionofAk t
[5]
pat
hwa ythroughS er308p hosph o
ryla
tions i
tes .Re sultsofimmu  prote
in,p r
omotedtheBa da ndc a
spase
9e xpres
sion,a nde n
nohi
stochemistrya ndWe sternBlo t
ti
ngs howe dt hatb lackg arl
icex hancedthee x
pres
sionofPTEN.Th er
efor
e,itc ou
ldb econcluded

racti
ngs ol
u t
ionh ade ff
ectso ntrans
p lant
edt u mo ro fmic ewithco th
a tPI3K/Aktpathwayofb l
ackg ar
li
ce xt
ract
ingsoluti
onh a
di n

onc ancerHt 29,a ndr est
rictedthee xpressiono fAk tp ro
tein. hi
b it
oryeff
ect
s,andHt 29mi c
etranspl
ante
dt umorwa srest
ri
c t
ed
Ba dbelon gedt ob cl
2f ami l
y,c ouldc omb inedwi thb cl2or th
ro u
gha dj
usti
ngth ePI3K/ Aktp a
thway.Ho wever,f ur
therre
bcl
xl,soa st oma keb cl
2a ndb cl
x llostitsa n ti
a po
p tos
isfunc searc
hwa ssti
llneededonthee f
fect
sofblackgarl
icex t
ract
ingso

ion,a ndtop romo tet hec el
la poptosis
.Th ep hosp horyl
atedBa d lu
tiono no t
herr e
lat
edp a
thways,o ront heirinte
racti
onwi th
wasma int
ainedi nt h ec yt
oplasma ndc oul
dn otb ec omb inedwi t
h PI3K/ Aktpat
hway.
bcl
2o rbclxl,s oa st oa voi
da p o
biosiso ftumo rc el
lsa ndt oac
chi
vep rol
ifer
ation[6-7].Th eref
ore,itwa so fg reats ig
n if
icanceto Ref
erences
det
ectt h
eBa de xpre s
sioni nt umort iss
u es.Att h es amet ime, [1]KI MS H,J UNGEY,KANGDH,e ta l.Ph ysicalstabili
ty,antioxida
tive
cas
p as
e9p layeda ni mp ort
antr ol
ei nt umo rc el
la poptos
is,a nd propert
ies,a ndp hot
o pro
tecti
vee f
fect
so fafu nct
ionali
zedf ormula
tionc on
tai
ningb l
ac kga r
li
ce xtract[J ].J ourna
lo fPh otochemistrya ndPh ot
o bi
ology
wasth estart
e ro fc e l
la poptosis.Ca s
pase 
9wa sc ata l
yzeda nd B:Bi ology,2 01
2 ,117C:1 04-1 10.
phosphory
latedb yAk ,s
t othatc as
pase 
9c ou l
dn ote xerttheapop [2]HUL,J IANGHG.Th estudyp ro
gressa bo
utme chani
smo fPI3K/ AKTs ig

osisp r
omo t
inge ffects.Th isresearchs howe dt hatb lackg arl
icex nalc o
n ducti
onp athwayr el
a t
edtot umorme tasta
sis[J ].Me dica
lRe ca
pitu

racti
ngs olut
ioni ncreasedt heBa dt ranscri
p tiona ndt ra
n s
lati
on l
a t
e,2006 ,12(22):13 7
5-1 373.(i nCh ine
se ).
[3]KURTZJ E,RAY COQUARDI .PI3k inasei n
hibitor
si nt h
ec li
nic:a nu p

evelint umo rtissue s,r educedt hee xpressiono fp hosphorylat
ed dat
e [J].An t
icancerRe search,201 2
,3 2 (
7 ):2463-2 470.
Bad,a nde nh ancedt hec aspase9e xpression.Th eref
o r
e,b l
ack [4]PAPADI MI TRAKOPOULOUV.De ve
lopme ntofPI 3K/ AKT/ mTORp at
hwa y
gar
licextra
c t
ings ol
u ti
onc ou l
dr est
rictt hep ro l
iferat
iono fHt 
29 i
n hi
b i
torsa n
dt heira pplic
a t
ioninp er
sonalize
dt h er
apyf o
rn onsma l
l
c e
ll

u ngc ancer[J].J ournalofTh or
acicOn cology,2 012,7 (8): 1
315-1 326.
cel
la ndh adtu mo rk il
linge f
fect
s.
[5]ALTOMAREDA,KHALEDAR.Ho me ost
asisandt hei mp o
rtanc
ef orab al
Asac ance rsu ppressorgener ecentl
yf oun d,PTENh adc lo
se anceb etweenAKT/ mTORa ctiv
ityandi ntr
a cel
lularsignali
n g[J].Cu r
rent
cor
relat
ionwi t
ht umo rd evel
op ment.Ab s
en to rre ducedPTENe x Med ic
inalCh emistr
y, 2 0
1 2,1 9(22):3748-3 7
62.
pr
e s
sionwa sfoun di nma nytumo rdis
e ases.Tu mo rc e
llh admi gr
a [6]LILQ,LIXL,WANGL,e ta l
.Ma t
rinein hi
bitsb r
ea s
tc ancergrowthv ia
miR21/ PTEN/ Ak tp ath
wa yi nMCF 7c ell
s[J ].Ce l
lularPh y
siol
o gya n


iona ndb oneme tasta
se s
,wh ichwa st hei mp ortantr easonwh y
Biochemis t
ry,2012,3 0 (3):6 31-6 41.

umo rwa sd i
ffi
c ulttoc onquer,a ndwa sth er esearchh ot
spotf o
r [7]QUANJ H,CHAGH,ZHOUW,e tal.Involvemen tofPI3k inas
e/ Aktde

umo rresear
ch.Re searchr es
u l
tsshowe dth atPTENc o
u ldregulat
e pendentBa dp hosphorylat
ioni nTo xoplas
mag ond i
imed i
atedinh i
b i
tono f
var
iouss i
gnalp a t
h wa y,in cl
ud i
ngPI 3K/ Ak t,J AK/ STAT,FAK hostcella popt
osis[J ].Ex p er
imentalPa r
asito
logy,2 013,1 3
3(4):4 62-
[8-9] 471.
andERK1/ 2 .Du ringthePI 3K/ Ak tsigna ltransdu ct
ionp at
h
[8]CHETRAM MA,HI NTONCV.PTENr e
gu la
ti
ono fERK1/ 2s i
gna l
ingin
way,e nhancingPTENe xpressi
onc ouldrestrictth ep hosphorylat
ed cancer[J ].J ournalofRe cep t
orsandS i
gnalTr ansduction,2 01
2,3 2(4):
Bada ndc aspase9i n ducedb yAk ,a
t nde xertt hea p opto
sispr
o 190-1 95.
moti
n geff
ectso fBa da ndc aspase9[10].
 [9]PANAGI OTOUI ,TS IAMBASE,LAZARI SA,e ta l.PTENe xpr
es si
onin
nons mallc el
llungc a
rcinomab asedo nd i
gitiz
edi magea n a
lysi
s[J ].J our
PI3K/ Ak tsign altransducti
onp athwayp arti
c i
p at
edi nt hede nalofB. U. ON. :o f
fici
aljournaloftheBa lkanUn iono fOn col
ogy,2 012,17
vel
opme ntproces so fma nyma li
gnantd ise
a ses,p romotedt hes ur
 (4): 7
1 9-7 23.
vi
val,p r
olif
eration,b l
o odv es
selforma ti
ona ndt u morme t
astasi
s, [10]HARAS, OYAM, MI ZUNOR, e
ta l.Ak ta ct
ivatio
ni nr enalcellc ar
cino
re
stri
ctedthePI 3K/ Ak tsignaltr
a n
sdu ct
ionp a t
h wa y,p romotedthe ma:c ontri
bu t
io
no fad ecreasedPTENe x
pre s
siona ndt heinducti
ono fa p
opt
osisb ya nAk tin hi
bito
r[J ].An nalsofOn cology,2005,16(6): 928.
cel
la poptos
is,i nhibit
e dp roli
fera
ti
ona nds oo n.Th isr es
e ar
ch
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(Frompa ge3) f
romGi nsen
gBa i
kalS ku
llc
apRo otontheinjuryoforgani
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