紫杉醇和多柔比星处理改变小鼠乳腺癌免疫微环境并抑制肿瘤细胞生长  

Treatment of paclitaxel and doxorubicin changes the immune microenvironment of breast cancer and inhibits the growth of tumor cells in mice

作　　者：王瑞 郎磊 陈善春 万雪颖 侯懿烜[3] WANG Rui;LANG Lei;CHEN Shanchun;WAN Xueying;HOU Yixuan(Key Laboratory of Clinical Laboratory Diagnostics of the Ministry of Education,Chongqing Medical University,Chongqing 400016;Clinical Laboratory of the Affiliated Central Hospital,Chongqing University,Chongqing 400014;Basic Medical Experimental Teaching Center,Chongqing Medical University,Chongqing 400016,China)

机构地区：[1]重庆医科大学临床检验诊断学教育部重点实验室,重庆400016 [2]重庆大学附属中心医院检验科,重庆400014 [3]重庆医科大学基础医学实验教学中心,重庆400016

出　　处：《细胞与分子免疫学杂志》2023年第10期891-897,共7页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81773078);重庆市自然科学基金(cstc2020jcyj-msxmX0845)。

摘　　要：目的探讨紫杉醇和多柔比星对小鼠乳腺癌免疫微环境的影响。方法用肿瘤药物应答相关基因表达谱及耐药特征分析数据库CTR-DB分析化疗药对乳腺癌免疫微环境的影响。用4T1细胞原位注射建立小鼠乳腺癌模型,分别用多柔比星和紫杉醇处理。测定小鼠肿瘤大小并绘制生长曲线。采用流式细胞术检测乳腺癌免疫细胞的数量,免疫组织化学染色法检测小鼠乳腺癌组织中乳腺癌抗原KI-67(ki67)、S100钙结合蛋白A9(S100A9)和基质金属蛋白酶9(MMP9)。流式细胞术检测紫杉醇组和多柔比星组中4T1细胞的周期。结果CTR_Microarray_75结果表明药物敏感的乳腺癌中免疫分值、细胞毒性淋巴细胞、B细胞谱系、CD8^(+)T细胞、树突状细胞(DC)、单核细胞谱系以及自然杀伤(NK)细胞等指标均比药物不敏感的乳腺癌高。通过乳腺癌小鼠生长曲线分析,发现紫杉醇和多柔比星均能显著抑制肿瘤体积的增长,而且紫杉醇的肿瘤抑制作用强于多柔比星。通过流式细胞术检测,我们发现紫杉醇和多柔比星均能抑制乳腺癌细胞ki67的表达,并使停滞在G2/M期的乳腺癌细胞增多,其中紫杉醇的作用明显强于多柔比星。多柔比星和紫杉醇均能导致CD45^(+)免疫细胞的增加和中性粒细胞的减少;而且紫杉醇诱导CD3^(+)CD4^(+)辅助性T细胞、CD3^(+)CD8^(+)细胞毒性T细胞、CD45^(+)CD19^(+)B细胞数量增加,而多柔比星诱导CD4^(+)CD25+调节性T细胞(Treg)增加。免疫组织化学染色结果表明紫杉醇对S100A9的抑制作用更强,而多柔比星对MMP9的抑制作用更强。结论紫杉醇和多柔比星可以有效抑制肿瘤细胞的生长,并调控不同的细胞浸润模式和不同细胞基质外成分,改变三阳性乳腺癌(TNBC)的免疫微环境。Objective To investigate the effects of paclitaxel and doxorubicin on the immune microenvironment of breast cancer in mice.Methods The CTR-DB database,a database for analysis of gene expression profiles and drug resistance characteristics related to tumor drug response,was used to analyze the effect of chemotherapeutic drugs on the immune microenvironment of breast cancer.Mouse models with breast cancer were established by in situ injection with 4T1 cells,a triple-negative breast cancer(TNBC)cells.Then they were treated with doxorubicin and paclitaxel,respectively.The sizes of tumor were recorded and analyzed by growth curve.The number of different types of immune cells was analyzed using flow cytometry.The expressions of Ki67,S100 calcium binding protein A9(S100A9)and matrix metalloproteinase 9(MMP9)were detected by immunohistochemistry.The cell cycles of 4T1 cells in paclitaxel group and doxorubicin group were analyzed by flow cytometry.Results The results of CTR_Microarray_75 analysis showed that the immune scores,and the number of cytotoxic lymphocytes,B lineages,CD8^(+)T cells,dendritic cells(DCs),monocytic lineages and natural killer(NK)cells in chemotherapy-sensitive breast cancer were higher than those in chemotherapy-insensitive breast cancer.Through growth curve analysis in mice with breast cancer,we found that both paclitaxel and doxorubicin could inhibit the increase of the tumor sizes,and the paclitaxel showed a higher inhibitory effect.The results of cytometry displayed that both paclitaxel and doxorubicin could restrain the expression of Ki67 and increase the number of breast cancer cells in G2/M phase,and in the paclitaxel group,the expression of Ki67 was lower and the number of breast cancer cells in G2/M phase was larger.Paclitaxel and doxorubicin enhanced the infiltration of CD45^(+)immune cells but decreased the infiltration of neutrophils.Additionally,paclitaxel promoted the infiltration of CD3^(+)CD4^(+)T helper cells,CD3^(+)CD8^(+)cytotoxic T cells and CD45^(+)CD19^(+)B cells,while doxorubi

关 键 词：紫杉醇 多柔比星 乳腺癌 免疫微环境 

分 类 号：R965[医药卫生—药理学] Q279[医药卫生—药学] R737.9[生物学—细胞生物学] S853.53[农业科学—临床兽医学] R392.1[农业科学—兽医学]