聚合人脐带血红蛋白氧载体增强乳腺癌裸鼠皮下移植瘤化疗敏感性的初步研究  

Preliminary study on polymerized human cord hemoglobin increasing the chemosensitivity of subcutaneous xenografts of breast cancer in nude mice

作　　者：高燚秋 朱珂慧 徐莹璨 索正新 王红[1] 刘嘉馨[1] GAO Yiqiu;ZHU Kehui;XU Yingcan;SUO Zhengxin;WANG Hong;LIU Jiaxin(Institute of Blood Transfusion,Chinese Academy of Medical Sciences and Peking Union Medical College,Chengdu 610051,China)

机构地区：[1]中国医学科学院北京协和医学院输血研究所,四川成都610052

出　　处：《中国输血杂志》2022年第5期475-479,共5页Chinese Journal of Blood Transfusion

基　　金：中国医学科学院医学与健康科技创新工程项目(2021-I2M-1-060);四川省科学技术厅项目(2021YJ0122)。

摘　　要：目的探讨聚合人脐带血红蛋白氧载体(PolyCHb)对人乳腺癌MCF-7细胞裸鼠皮下移植瘤化疗敏感性的影响及其机制。方法收集处于指数生长期的MCF-7细胞,制成密^度为5×10^(7)个/mL的悬浮细胞,按0.2 mL/只接种于18只BALB/c-nu裸鼠右肢皮下,建立裸鼠移植瘤模型,待肿瘤体积达到100 mm^(3)左右时,随机均分为化疗组:多柔比星(DOX)5 mg·kg^(-1),1次/周;化疗+PolyCHb治疗组:除给药(DOX)(同化疗组)外,PolyCHb 600 mg·kg^(-1),3次/周;对照组:生理盐水90 mg·kg^(-1),1次/周;均为经尾静脉连续注射4周。自注射当天(d0)起,每3 d 1次测量各组裸鼠肿瘤体积,据此绘制其各自(组)肿瘤生长曲线。38 d结束肿瘤生长观察,剥瘤并称取瘤重,计算抑瘤率;HE染色观察肿瘤组织病理变化,免疫组化检测肿瘤组织中HIF-1α表达,采用TUNEL法检测肿瘤细胞凋亡情况,荧光染色测定各组肿瘤组织活性氧(ROS)的含量。结果化疗+PolyCHb组、化疗组和对照组至d38时的肿瘤体积(mm^(3))分别为:196.35±103.45 vs 316.29±62.88 vs 519.42±177.33(P<0.05);化疗+PolyCHb组与化疗组抑瘤率(%)分别为62.20 vs 39.11;HE染色和TUNEL检测:化疗+PolyCHb组肿瘤组织生长区域细胞坏死和凋亡增多;免疫组化:化疗+PolyCHb组HIF-1α表达水平降低;荧光染色:化疗+PolyCHb组[活性氧(ROS)]含量增多。结论PolyCHb增加了乳腺癌裸鼠皮下移植瘤化疗的敏感性,其作用机制可能与其提高肿瘤组织内ROS含量,促进肿瘤细胞的凋亡有关。Objective To investigate the effect of polymerized human cord hemoglobin(PolyCHb) on the chemosensitivity of human breast cancer MCF-7 cell subcutaneous xenografts in nude mice and its mechanism.Methods The MCF-7 cells in exponential growth phase were collected and made into suspension cells at a density of 5×10^(7) cells/mL.Subsequently, the cells were inoculated subcutaneously in the right limb of 18 BALB/c-nu nude mice with 0.2 mL cells per mouse to establish subcutaneous xenograft.When the tumor volume reached about 100 mm^(3),they were randomly divided into chemotherapy group: doxorubicin 5 mg·kg^(-1),once/week;chemotherapy + PolyCHb group: in addition to doxorubicin(chemotherapy group),PolyCHb 600 mg·kg^(-1),3 times/week;the control group: normal saline 90 mg·kg^(-1),once/week;all were injected through tail vein continuously for 4 weeks.From the day of injection(d 0),the tumor volume of each group of nude mice was measured every 3 days, and the tumor growth curves were drawn accordingly.After 38 days, the tumor growth observation was completed.The tumor was removed and weighed to calculate the tumor inhibition rate.HE staining, immunohistochemistry and TUNEL method were used to observe the pathological changes of tumor tissue, detect the expression of HIF-1α,and detect tumor cell apoptosis respectively.The content of reactive oxygen species(ROS) of each group was determined by fluorescence staining.Results The tumor volume(mm~3) of chemotherapy + PolyCHb group, chemotherapy group and the control group at day 38 were 196.35±103.45 vs 316.29±62.88 vs 519.42±177.33(P<0.05),and the tumor inhibition rate(%) of chemotherapy + PolyCHb treatment group and chemotherapy group was 62.20 vs 39.11,respectively.HE staining and TUNEL detection showed that cell necrosis and apoptosis in the growth area of tumor tissue increased in chemotherapy +PolyCHb group.Immunohistochemistry and fluorescence staining showed that HIF-1α expression in chemotherapy +PolyCHb group decreased and reactive oxygen species(ROS) content

关 键 词：聚合人脐带血红蛋白 皮下移植瘤动物模型 乳腺癌 化疗敏感性 活性氧 DOX 裸鼠 动物实验 

分 类 号：R457.1[医药卫生—治疗学]