负载紫杉醇的丝素蛋白纳米粒子对乳腺癌荷瘤小鼠肿瘤生长及凋亡蛋白表达的影响  

Effects of silk fibroin nanoparticles loaded with paclitaxel on tumor growth and expression of apoptosis protein in breast cancer-bearing mice

作　　者：夏凡林 沈磊莹 曹阳 林子晶 XIA Fanlin;SHEN Leiying;CAO Yang;LIN Zijing(Shanghai Urban Construction Vocational College,Shanghai 201415,China)

机构地区：[1]上海城建职业学院健康与社会关怀学院,上海201415 [2]上海交大医学院附属第九人民医院 [3]重庆医科大学附属第二医院

出　　处：《山东医药》2021年第2期12-16,共5页Shandong Medical Journal

基　　金：重庆市博士后科研项目(Xm2015089)。

摘　　要：目的构建负载紫杉醇的丝素蛋白纳米粒子,观察其对乳腺癌荷瘤小鼠肿瘤生长及凋亡蛋白表达的影响。方法从蚕丝纤维中提取丝素蛋白,加入液体石蜡中乳化,戊二醛固化,冷冻干燥后获得空白微球。将紫杉醇加入乙醇溶液完全溶解后,按照制备空白微球的方法制备载药微球。向小鼠右前肢腋皮下注射乳腺癌EMT6细胞悬液,建立乳腺癌荷瘤小鼠模型。将造模成功的80只小鼠随机分为对照组、紫杉醇组、紫杉醇载体组、空白微球组各20只。紫杉醇组于接种后第8、11、14天给予紫杉醇5 mg/kg腹腔注射,紫杉醇载体组于相同时间点给予与紫杉醇组同等药量的载药微球溶液腹注射腔,空白微球组注射等体积空白微球溶液,对照组注射等体积生理盐水。造模后每5 d行B超检查1次,测量瘤体体积,计算抑瘤率;造模第22天采集眼眶静脉血,使用全自动生化分析仪测定血清ALT、AST、BUN、肌酐水平;采用高效液相色谱法检测肿瘤组织中紫杉醇药物浓度;流式细胞术检测组织中肿瘤细胞凋亡率;Western blotting法检测肿瘤组织中凋亡相关蛋白cleaved Caspase-3、Bax、Bcl-2表达。结果紫杉醇载体组抑瘤率高于紫杉醇组(P<0.05)。紫杉醇组、紫杉醇载体组及空白微球组血清肝肾功能与对照组比较差异均无统计学意义(P均>0.05)。紫杉醇载体组肿瘤组织中的紫杉醇浓度高于紫杉醇组(P<0.05)。紫杉醇载体组、紫杉醇组细胞凋亡率均高于对照组和空白微球组,紫杉醇载体组肿瘤细胞凋亡率高于紫杉醇组(P均<0.05)。与对照组和空白微球组相比,紫杉醇载体组、紫杉醇组肿瘤组织中cleaved Caspase-3、Bax蛋白表达升高,Bcl-2表达降低(P均<0.05);紫杉醇载体组cleaved Caspase-3及Bax蛋白表达均高于紫杉醇组,Bcl-2表达低于紫杉醇组(P均<0.05)。结论负载紫杉醇的丝素蛋白纳米粒子可提高肿瘤组织中的紫杉醇浓度,促进肿瘤细胞凋亡,�Objective To construct silk fibroin nanoparticles loaded with paclitaxel and to observe their effects on tumor growth and apoptosis protein expression in breast cancer bearing mice.Methods The silk fibroin was extracted from silk fiber,and then we added it to liquid paraffin for emulsification,glutaraldehyde for solidification,and obtained blank microspheres after freeze-drying.After paclitaxel was added to the ethanol solution to completely dissolve,the drugloaded microspheres were prepared according to the method of preparing blank microspheres.The breast cancer EMT6 cell suspension was injected subcutaneously into the right forelimb axillary of the mouse to establish the breast cancer-bearing mouse model.The 80 successfully modeled mice were randomly divided into the control group,paclitaxel group,paclitaxel carrier group,and blank microsphere group,with 20 mice in each.Mice in the paclitaxel group received 5 mg/kg intraperitoneal injection of paclitaxel on day 8,11,and 14 after vaccination.Mice in the paclitaxel carrier group received intraperitoneal injection of paclitaxel at the same time with a calculated concentration of 5 mg/kg drug-loaded microsphere solu‑tion.In the blank microsphere group,equal volume of blank microsphere solution was injected,and the mice in the control group were injected with an equal volume of normal saline.The concentration of paclitaxel in the tumor tissues was detected by high performance liquid chromatography(HPLC)method.The mouse orbital venous blood was collected and the serum ALT,AST,BUN,and creatinine levels were detected.The average tumor weight were recorded and tumor inhibition rate was calculated.Apoptosis rate of tumor cells was detected by flow cytometry.The expression of apoptosis-related proteins cleaved Caspase-3,Bax,and Bcl-2 in the tumor tissues were detected by Western blotting.Results The tumor growth inhibitory rate of the paclitaxel carrier group was higher than that of the paclitaxel group(P<0.05).No significant differences were found in the serum liver o

关 键 词：乳腺癌 紫杉醇 丝素蛋白 Caspase-3 Bax Bcl-2 小鼠 

分 类 号：R737.9[医药卫生—肿瘤]