叶酸偶联白蛋白Cypate纳米粒介导光热治疗效应对荷人膀胱癌5637细胞裸鼠的抑瘤作用及其机制  

Photothermal therapy with folic acid-bovine serum albumin-Cypate nanoparticles on bladder cancer 5637 cells in vitro and the antitumor effect and mechanism of tumor-bearing nude mice in vivo

作　　者：朱伟[1] 朱庆国[1] Zhu Wei;Zhu Qingguo(Department of Urology,Fujian Provincial Hospital,Shengli Clinical Medical Collge of Fujian Medical University,Fuzhou 350001,China)

机构地区：[1]福建医科大学省立临床医学院福建省立医院泌尿外科,福州350001

出　　处：《中华实验外科杂志》2021年第6期1032-1036,共5页Chinese Journal of Experimental Surgery

基　　金：福建省自然科学基金面上项目(2017J01251)。

摘　　要：目的:探讨FA-BSA-Cypate纳米粒介导的靶向PTT效应对人膀胱癌5637细胞荷瘤裸鼠的抑瘤作用及其机制。方法:免疫荧光法检测5637细胞叶酸(FA)受体分布;1,3-二苯基异苯并呋喃(DPBF)消耗量检测单态氧(1O 2)生成。分4组:空白对照组(N组)、激光组(L组)、BSA-Cypate+激光组(BL组)、FA-BSA-Cypate+激光组(FL组)。流式细胞术检测各组细胞凋亡率;蛋白质印迹法(Western blot)法测定热休克蛋白(Hsp)70、Hsp90蛋白表达变化;建立裸鼠膀胱癌5637细胞皮下移植瘤模型,观察FA-BSA-Cypate纳米粒介导的体内靶向抗肿瘤效应。应用GraphPad Prism 5.0统计软件,采用单因素方差分析及t检验。结果:人膀胱癌5637细胞膜表达叶酸受体;808 nm激光照射,FA-BSA-Cypate纳米粒能促进1O 2生成。FL、BL、L及N组细胞凋亡率分别为(56.912±5.457)%、(43.941±4.097)%、(12.923±2.489)%及(8.401±1.453)%,FL组与其他各组比较差异有统计学意义(F=203.502,P<0.05)。FL、BL、L及N组Hsp70、Hsp90蛋白相对表达分别是1.903±0.097及4.833±0.208;1.410±0.123及3.731±0.153;0.910±0.224及2.501±0.265;0.867±0.153及2.533±0.252;FL组与其他各组比较差异有统计学意义[F=25.180(Hsp70),P<0.05;F=74.530(Hsp90),P<0.05];荷瘤裸鼠模型提示FL、BL、L及N组肿瘤区域最高温度分别为(58.512±1.936)、(50.334±1.483)、(38.260±2.177)、(34.322±1.308)℃,至第21天肿瘤相对体积分别是7.462±0.462、5.581±0.669、1.403±0.354、0.386±0.089,FL组较其他各组表现出更显著的靶向光热效应(F=198.612,P<0.05)及抗肿瘤增殖作用(F=285.403,P<0.05),差异均有统计学意义。结论:FA-BSA-Cypate纳米粒介导的PTT效应对人膀胱癌5637细胞具有显著的靶向杀伤效果,促进了5637细胞的凋亡及单线态氧的生成;FA-BSA-Cypate纳米粒介导的光热疗法对人膀胱癌5637细胞荷瘤裸鼠有抑瘤效果。Objective To investigate the antitumor effect of targeted photothermal therapy(PTT)mediated by FA-BSA-Cypate nanoparticles on human bladder cancer 5637 cell tumor-bearing nude mice and its potential mechanism.Methods The distribution of folate receptor in 5637 cells was detected by immunofluorescence assay.1,3-diphenylisobenzofuran(DPBF)consumption was detected for singlet oxygen(1O2)generation.The cells were divided into four groups,control group(group N),laser group(group L),BSA-Cypate+laser group(group BL),and folic acid-bovine serum albumin(FA-BSA)-Cypate+laser group(group FL).The apoptosis rate of each group was detected by flow cytometry.The protein expression of heat-shock protein 70(Hsp70)and Hsp90 was detected by Western blotting.The subcutaneous tumor transplantation model of nude mouse bladder cancer 5637 cells was established to observe the targeted antitumor effect mediated by FA-BSA-Cypate nanoparticles in vivo.GraphPad Prism 5.0 statistical software,One way ANOVA and t test were used for statistical analysis.Results The cell membrane of human bladder cancer 5637 cells expressed folate receptor.Under 808nm laser irradiation,FA-BSA-Cypate nanoparticles could promote singlet oxygen generation.The apoptosis rate of in groups FL,BL,L and N was(56.912±5.457)%,(43.941±4.097)%,(12.923±2.489)%and(8.401±1.453)%respectively.The apoptosis rate in group FL was significantly higher than that in the rest groups(F=203.502,P<0.05).The expression of Hsp70 and Hsp90 in groups FL,BL,L and N was 1.903±0.097 and 4.833±0.208,1.410±0.123 and 3.731±0.153,0.910±0.224 and 2.501±0.265,0.867±0.153 and 2.533±0.252 respectively.The expression of Hsp70 and Hsp90 was significantly increased in the group FL[F=25.180(Hsp70),P<0.05;F=74.530(Hsp90),P<0.05].The tumor-bearing nude mouse model suggested that the maximum temperature in groups FL,BL,L and N was(58.512±1.936),(50.334±1.483),(38.260±2.177)and(34.322±1.308)℃respectively,and the relative TV in groups FL,BL,L and group N was 7.462±0.462,5.581±0.669,1.403±0.

关 键 词：膀胱肿瘤 治疗 纳米技术 

分 类 号：R737.14[医药卫生—肿瘤]