三阴乳腺癌(Triple Negative Breast Cancer,TNBC)是乳腺癌中恶性程度最高的一种亚型,表现为很高的转移潜能。巨噬细胞,即肿瘤相关巨噬细胞(Tumor-Associated Macrophages,TAM),在促进TNBC转移中起了重要作用。乳腺癌作为一种实体肿瘤,...三阴乳腺癌(Triple Negative Breast Cancer,TNBC)是乳腺癌中恶性程度最高的一种亚型,表现为很高的转移潜能。巨噬细胞,即肿瘤相关巨噬细胞(Tumor-Associated Macrophages,TAM),在促进TNBC转移中起了重要作用。乳腺癌作为一种实体肿瘤,往往处于缺氧环境中。低氧环境能够促进癌细胞的转移,然而低氧环境中巨噬细胞在促进肿瘤转移中的作用仍然不清楚。在该研究中,THP1细胞被诱导成TAM,经过缺氧培养后,通过Transwell实验检测其促进三阴乳腺癌细胞BT-549和MDA-MB-231的细胞迁移能力;通过尾静脉注射,将MDA-MB-231细胞移植于祼鼠体内,CT扫描,分析了TAM促进TNBC细胞的器官转移能力;通过ELISA实验检测低氧对TAM分泌的肿瘤转移相关因子的影响,通过GDSC在线软件分析了CCL22受体CCR4和其他CCR在乳腺癌组织与正常组织中表达的差异。结果表明低氧条件下巨噬细胞通过分泌CCL22的表达来促进三阴乳腺癌细胞迁移:经过缺氧培养后的TAM显著增强了TNBC细胞迁移能力,以及促进癌细胞在体内向肺转移;低氧诱导TAM分泌CCL22;CCL22受体CCR4在乳腺癌组织中的表达显著高于在正常组织中的。展开更多
Objective:To explore the mechanism by which macrophages secrete CCL22 to promote the metastasis of triple-negative breast cancer under hypoxic conditions.Methods:20 ng/mL mass concentration of phorbol 12-myristate 13-...Objective:To explore the mechanism by which macrophages secrete CCL22 to promote the metastasis of triple-negative breast cancer under hypoxic conditions.Methods:20 ng/mL mass concentration of phorbol 12-myristate 13-acetate(PMA)cell culture medium,4′,6-diamidino-2-phenylindole(DAPI),dimethyl sulfoxide(DMSO),trypsin digestion solution,CCL18 Kit,Interleukin(IL)-10 Kit,CCL17 Kit,CCL22 Kit,TRIzol^(TM)Reagent Reverse Transcriptase Polymerase Chain Reaction(RT-PCR)Kit,triple-negative breast cancer cells MDA-MB-231 and BT-549,as well as other reagents were used to culture triple-negative breast cancer(TNBC)cells MDA-MB-231 and BT-549 as well as human mononuclear cells THP-1,analyze and observe the metastasis of triple-negative breast cancer cells to the lungs,the secretion of CCL22,the migration of triple-negative breast cancer,and the situation of CCR4.Results:Compared with normal tumor-associated macrophage(TAM),hypoxic TAM further promoted the migration of tumor cells.The number of tumor metastases in the lungs,induced by hypoxic TAM,was significantly higher than that of normoxic TAM.Hypoxia can significantly stimulate the expression of CCL22.CCL22 can significantly promote the migration of MDA-MB-231 and BT-549 cells.The expressions of CCR3,CCR4,and CCR5 in tumor tissues were significantly increased compared with normal tissues,in which CCR4 showed the most significant increase.Conclusion:TAM cultured under hypoxia significantly enhanced the migration ability of TNBC cells and promoted the metastasis of cancer cells to the lungs in vivo.The hypoxic condition induced TAM to secrete CCL22;the expression of CCL22 receptor,CCR4,in breast cancer tissues was significantly higher than that in normal tissues.展开更多
文摘三阴乳腺癌(Triple Negative Breast Cancer,TNBC)是乳腺癌中恶性程度最高的一种亚型,表现为很高的转移潜能。巨噬细胞,即肿瘤相关巨噬细胞(Tumor-Associated Macrophages,TAM),在促进TNBC转移中起了重要作用。乳腺癌作为一种实体肿瘤,往往处于缺氧环境中。低氧环境能够促进癌细胞的转移,然而低氧环境中巨噬细胞在促进肿瘤转移中的作用仍然不清楚。在该研究中,THP1细胞被诱导成TAM,经过缺氧培养后,通过Transwell实验检测其促进三阴乳腺癌细胞BT-549和MDA-MB-231的细胞迁移能力;通过尾静脉注射,将MDA-MB-231细胞移植于祼鼠体内,CT扫描,分析了TAM促进TNBC细胞的器官转移能力;通过ELISA实验检测低氧对TAM分泌的肿瘤转移相关因子的影响,通过GDSC在线软件分析了CCL22受体CCR4和其他CCR在乳腺癌组织与正常组织中表达的差异。结果表明低氧条件下巨噬细胞通过分泌CCL22的表达来促进三阴乳腺癌细胞迁移:经过缺氧培养后的TAM显著增强了TNBC细胞迁移能力,以及促进癌细胞在体内向肺转移;低氧诱导TAM分泌CCL22;CCL22受体CCR4在乳腺癌组织中的表达显著高于在正常组织中的。
基金supported by grants from The Medical Science Research Project of Hebei Province(20220282)the Key Laboratory of Molecular Pathology and Early Diagnosis of Tumor in Hebei Province.
文摘Objective:To explore the mechanism by which macrophages secrete CCL22 to promote the metastasis of triple-negative breast cancer under hypoxic conditions.Methods:20 ng/mL mass concentration of phorbol 12-myristate 13-acetate(PMA)cell culture medium,4′,6-diamidino-2-phenylindole(DAPI),dimethyl sulfoxide(DMSO),trypsin digestion solution,CCL18 Kit,Interleukin(IL)-10 Kit,CCL17 Kit,CCL22 Kit,TRIzol^(TM)Reagent Reverse Transcriptase Polymerase Chain Reaction(RT-PCR)Kit,triple-negative breast cancer cells MDA-MB-231 and BT-549,as well as other reagents were used to culture triple-negative breast cancer(TNBC)cells MDA-MB-231 and BT-549 as well as human mononuclear cells THP-1,analyze and observe the metastasis of triple-negative breast cancer cells to the lungs,the secretion of CCL22,the migration of triple-negative breast cancer,and the situation of CCR4.Results:Compared with normal tumor-associated macrophage(TAM),hypoxic TAM further promoted the migration of tumor cells.The number of tumor metastases in the lungs,induced by hypoxic TAM,was significantly higher than that of normoxic TAM.Hypoxia can significantly stimulate the expression of CCL22.CCL22 can significantly promote the migration of MDA-MB-231 and BT-549 cells.The expressions of CCR3,CCR4,and CCR5 in tumor tissues were significantly increased compared with normal tissues,in which CCR4 showed the most significant increase.Conclusion:TAM cultured under hypoxia significantly enhanced the migration ability of TNBC cells and promoted the metastasis of cancer cells to the lungs in vivo.The hypoxic condition induced TAM to secrete CCL22;the expression of CCL22 receptor,CCR4,in breast cancer tissues was significantly higher than that in normal tissues.
