激光生物学报
激光生物學報
격광생물학보
ACTA LASER BIOLOGY SINICA
2014年
1期
16-24
,共9页
刘忠涛%刘志鹏%熊力%苗雄鹰%林良武%文宇
劉忠濤%劉誌鵬%熊力%苗雄鷹%林良武%文宇
류충도%류지붕%웅력%묘웅응%림량무%문우
空心纳米化光敏剂%光动力疗法%肝癌%细胞凋亡
空心納米化光敏劑%光動力療法%肝癌%細胞凋亡
공심납미화광민제%광동력요법%간암%세포조망
nanoscale photosensitize%conventional photosensitizer%photodynamic therapy%hepatic carcinoma%cell apoptosis
目的:探讨空心纳米粒子光敏剂与普通光敏剂在不同实验条件下对人HepG2肝癌细胞增殖的抑制作用及其作用机制。方法:应用体内外培养的HepG2肝癌细胞,以纳米化的photosan及photosan为光敏剂,半导体激光器(波长630 nm)为光源进行光照射,在不同的浓度光敏剂经不同剂量的光照后,用MTT法测定光动力治疗后肝癌细胞的残存率,并用流式细胞分析检测光动力治疗对肝癌细胞凋亡的影响,采取Western blot检测光动力治疗肝癌后凋亡相关蛋白caspase-3和caspase-9的表达情况,并建立裸鼠动物模型,比较纳米粒子光敏剂与普通光敏剂在动物体内对肝癌抑制作用。结果:纳米粒子光敏剂介导的光动力治疗对肝癌有明显的抑制作用,在一定范围条件下,随着光敏剂浓度的增加和激光剂量的的增大,肝癌细胞的的残存率明显下降,且纳米粒子光敏剂在同等条件下要优于普通光敏剂,流式细胞分析显示,空心纳米粒子光敏剂光动力治疗肝癌出现了明显的凋亡,且比普通光敏剂更显著。Western blot检测结果显示,两种光敏剂均引起凋亡蛋白caspase-3和caspase-9的表达,在空心纳米粒子光敏剂光动力作用组该两种蛋白的表达水平要明显高于普通光敏剂组。体内实验显示:用纳米粒子光敏剂与普通光敏剂分别对裸鼠肝癌模型干预,在生存期,瘤体的体积大小,纳米粒子光敏剂组显然优越于普通光敏剂组。结论:空心纳米粒子光敏剂对肝癌细胞在体内外具有明确的抑制作用,这种抑制作用可能通过引起凋亡相关蛋白高水平的表达,从而促发凋亡通路的开放,引起癌细胞发生凋亡。
目的:探討空心納米粒子光敏劑與普通光敏劑在不同實驗條件下對人HepG2肝癌細胞增殖的抑製作用及其作用機製。方法:應用體內外培養的HepG2肝癌細胞,以納米化的photosan及photosan為光敏劑,半導體激光器(波長630 nm)為光源進行光照射,在不同的濃度光敏劑經不同劑量的光照後,用MTT法測定光動力治療後肝癌細胞的殘存率,併用流式細胞分析檢測光動力治療對肝癌細胞凋亡的影響,採取Western blot檢測光動力治療肝癌後凋亡相關蛋白caspase-3和caspase-9的錶達情況,併建立裸鼠動物模型,比較納米粒子光敏劑與普通光敏劑在動物體內對肝癌抑製作用。結果:納米粒子光敏劑介導的光動力治療對肝癌有明顯的抑製作用,在一定範圍條件下,隨著光敏劑濃度的增加和激光劑量的的增大,肝癌細胞的的殘存率明顯下降,且納米粒子光敏劑在同等條件下要優于普通光敏劑,流式細胞分析顯示,空心納米粒子光敏劑光動力治療肝癌齣現瞭明顯的凋亡,且比普通光敏劑更顯著。Western blot檢測結果顯示,兩種光敏劑均引起凋亡蛋白caspase-3和caspase-9的錶達,在空心納米粒子光敏劑光動力作用組該兩種蛋白的錶達水平要明顯高于普通光敏劑組。體內實驗顯示:用納米粒子光敏劑與普通光敏劑分彆對裸鼠肝癌模型榦預,在生存期,瘤體的體積大小,納米粒子光敏劑組顯然優越于普通光敏劑組。結論:空心納米粒子光敏劑對肝癌細胞在體內外具有明確的抑製作用,這種抑製作用可能通過引起凋亡相關蛋白高水平的錶達,從而促髮凋亡通路的開放,引起癌細胞髮生凋亡。
목적:탐토공심납미입자광민제여보통광민제재불동실험조건하대인HepG2간암세포증식적억제작용급기작용궤제。방법:응용체내외배양적HepG2간암세포,이납미화적photosan급photosan위광민제,반도체격광기(파장630 nm)위광원진행광조사,재불동적농도광민제경불동제량적광조후,용MTT법측정광동력치료후간암세포적잔존솔,병용류식세포분석검측광동력치료대간암세포조망적영향,채취Western blot검측광동력치료간암후조망상관단백caspase-3화caspase-9적표체정황,병건립라서동물모형,비교납미입자광민제여보통광민제재동물체내대간암억제작용。결과:납미입자광민제개도적광동력치료대간암유명현적억제작용,재일정범위조건하,수착광민제농도적증가화격광제량적적증대,간암세포적적잔존솔명현하강,차납미입자광민제재동등조건하요우우보통광민제,류식세포분석현시,공심납미입자광민제광동력치료간암출현료명현적조망,차비보통광민제경현저。Western blot검측결과현시,량충광민제균인기조망단백caspase-3화caspase-9적표체,재공심납미입자광민제광동력작용조해량충단백적표체수평요명현고우보통광민제조。체내실험현시:용납미입자광민제여보통광민제분별대라서간암모형간예,재생존기,류체적체적대소,납미입자광민제조현연우월우보통광민제조。결론:공심납미입자광민제대간암세포재체내외구유명학적억제작용,저충억제작용가능통과인기조망상관단백고수평적표체,종이촉발조망통로적개방,인기암세포발생조망。
Objective:This study examined the inhibitory effects of conventional photosensitizers and photosensitizers delivered in hollow silica nanoparticles on the proliferation of HepG2 human hepatoma cells under different experimental conditions and the underlying mechanisms of these effects. Methods:Photosensitizers (conventional Photosan or nanoscale Photosan)were administered to in vivo and in vitro cultured HepG2 hepatoma cells,and a semiconductor laser was used as a light source to photoirradiate these cells.Photodynamic therapies(PDTs)involving different levels of light exposure and different photosensitizer concentrations were tested;To assess the effects of these PDTs,the cellular viabil-ity was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyl tetrazolium bromide (MTT)assay.Apoptotic and necrotic cells were measured by flow cytometry.The express of caspase-3 and caspase-9 caused in the photodynamic therapy by western blot.A nude mouse animal model was established to conduct in vivo experiments comparing the in-hibitory effects of nanoscale photosensitizers and conventional photosensitizers on liver cancer.Results:Nanoscale photo-sensitizer-mediated PDTs produced significant inhibitory effects on liver cancer cells.Within a certain range of condi-tions,hepatoma cell viability significantly decreased as photosensitizer concentrations and laser doses increased.Moreo-ver,under the same experimental conditions,the nanoscale photosensitizer performed better than the conventional photo-sensitizer(P<0.05).The flow cytometry demonstrated that the laser induced cell death with PS-loaded HSNP was much more severe than that of free PS (P<0.05 ).The activated forms the Caspase3 (Caspase3 20 kD)and caspase-9 (Caspase 9 35 kD)expression with PS-loaded HSNP were significantly higher than free PS(P<0.05 ).In vivo experi-ments using either nanoscale photosensitizer or conventional photosensitizer in a nude mouse liver cancer model suggested that nanoscale photosensitizer treatments were superior to the corresponding conventional photosensitizer treatments with re-spect to survival time and tumor volume.Conclusions:Hollow nanoparticles containing photosensitizer clearly inhibited hepatoma cells both in vivo and in vitro.These inhibitory effects may result from the induction of high levels of apoptosis-re-lated protein expression,which trigger the activation of apoptotic pathways that cause the programmed death of cancer cells.