·230·
中国肿瘤临床
2009年第36卷第8期
·基础研究·
新型光敏剂CPD4对人乳腺癌细胞体外光动力杀伤效应:l:
摘要
目的:研究新型光敏荆叶绿素衍生物4(chlomphyU
dedvative,CPD4)对人乳腺癌细胞系T47D和MCF一7
体外光动力杀伤效应,并对机理进行初步探讨。方法:采用激光共聚焦荧光显微镜结合荧光探针标记技术观察光
敏剂在细胞内分布及亚细胞定位;将终浓度分别为2.5、2.0、1.5、1.O、O.5斗咖L的CPD4加入到T47D及MCF-7细胞
培养皿中,孵育2h后用670nm半导体激光治疗仪照射,光剂量为10J,cm2,照毕,继续培养24h后采用染料排斥实验
测定细胞的存活率。流式细胞仪AnnexinV—FlrI’C,Pl双染法检测细胞死亡方式和凋亡率。实验分为四组:空白对
照组、单纯光敏剂组、单纯细胞光照组,光敏剂一细胞光照组。结果:光敏荆CPD4可穿过T47D及MCF-7细胞膜进入
细胞,分布在细胞膜及胞浆中,线粒体是其亚细胞分布的主要住点之一。光动力组中光敏剂浓度分别为2.5、2.O、
1.5、1.0、0.5斗g/mL,T47D及McF一7细胞死亡率分别为(76.77±6.41)%、(40.51±7.28)%、(38.4±5.35)%、(17.08±
0.52)%、(“.16±O.28)%、(81.67士1.53)%、(69.67±4.51)%、(45.33±4.73)%、(17.67±2.52)%、(12.00±2.oo)%。
Annexinv—FITc/PI法检测PDT作用24h后凋亡率分别可达36.80%及40.90%。结论:新型光敏荆CPD4分布在人乳
腺癌细胞T47D及MCF-7的细胞膜及细胞浆中,线粒体是其亚细胞分布主要位点之一;光敏剂CPD4对人乳腺癌细
胞T47D和McF一7具有良好的光动力杀伤效应,呈现量效关系。诱导细胞凋亡是其主要杀伤方式。线粒体途径促进
细胞凋亡可能是其光动力杀伤机制。
关键词光敏剂CPD乳腺癌光动力疗法
doi:lO.3969,j.issn.1000—8179.2009.08.014
Photodvnamic
Effects
of
New
Photosensitizer
CPD4
for
Human
Breast
Cancer
Cells
in
Vitro
QIAO
Zhu,CHEN
Ping,TONG
Zhongsheng,ZHANG
Siwen,LIN
Lie,LI
Shul.en
Corresponding
author:TONG
Zhongsheng,E—mail:tonghang@medmail.com.cn
Department
0f
Breast
Cance
r,Cancer
Inst.tute
and
HospitaI
of
11anjin
MedicaI
Universjty。11anjin
300060,Chi—
na
lnstitute
0f
Modem
Optics。Nankai
UniVerSi坝The
Key
Laborato叫of
Opt-eIectmnic
Informatlon
Science
and
。rechnoIogy,11anjin
300071。China
Grant
support:NationaI
NaturaI
Science
Foundation
of
Chlna(60508004):National
NaturaI
Scjence
Founda-
tion
of
China(60778043):Ph.D.Programs
Foundatjon
of
Ministry
of
Education
of
China(No.2006055024)
Abstract
obj∞Hve:1-0
investigate
the
photodynamic
e仟ects
of
CPD舢mediated
photodynamic
the阳py
0n
breast
cancer
ce¨Iine
T47D
and
MCF一7,and
t0
discuss
the
pOssibIe
mechanism.Meth似Is:Human
b怆ast
cancer
ce¨s
of
the
line‘r47D
were
cO-cuItured
w.th
CPD4。Rhodamine-1
23,and
mitOchOndria啊uOresCence
probe
dyes.Laser
scanning
confDcaI翻uOrescence
micrOscOpe
was
app¨ed
to
co¨ect
the
fIuorescence
of
the
phOtOsensItizer
and
the
probes
and
tO
image
the
ce¨s.T47D
and
MCF一7
ce¨s
were
cuItured
and
divided
Int0
4
groups
incIuding
the
bIank∞ntrol
group,the
Iaser
irrijdiation
grOup
wIthout
phOtosensitize
r,the
photosensi—
tizer
grOup
withOut
irradiation,and
the
PDT
gnDup.Dye
excIusiOn
method
was
app¨ed
t0
detect
the
death
rate
of
T47D
and
MCF.7
ce¨s.ApoptOtjc
ratiO
and
the
characteristics
of
ce¨death
was
anaIyzed
by
Annexin
V二FlTC,PI
doubIe
labe¨ed
staining
assay.R船ults:The伺uorescence
Of
the
photosensitizer
and
that
0f
the
probes
we旧0bserved
in
the
ce¨memb旧ne
and
cytopIasm
and
in
the
pe
rj—nucIear
region,mainIy
in
the
mlto—
chondria
0f
the
T47D
and
MCF.7
ceIIs.The
death阳te
0f
T47D
and
MCF一7
ce¨s
t陀ated
wIth
2.5,2.0,1.5,1.0,
·本文课题受国家自然科学基金(编号:60778043)和教育部博士点基金资助(编号:20060055024)
①南开大学现化光学研究所光电信息技术科学教育部重点实验室
通讯作者:佟仲生
tongllang@medmail.c叫.cn
万方数据