自然杀伤细胞联合拉帕替尼对乳腺癌细胞体外杀伤作用及机制研究  

Research on cytotoxicity and mechanisms of natural killer cell scombined with lapatinib on breast cancer cells in vitro

作　　者：孙越 杨琳 王文榜 张志斐 金媛媛[2] 于德红 杨兆勇[2] SUN Yue;YANG Lin;WANG Wen-bang;ZHANG Zhi-fei;JIN Yuan-yuan;YU De-hong;YANG Zhao-yong(College of Pharmacy,North China University of Science and Technology,Tangshan 063210,China;Institute of Medical Biotechnology,Chinese Academy of Medical Sciences,Beijing 100050,China)

机构地区：[1]华北理工大学药学院,唐山063210 [2]中国医学科学院/北京协和医学院医药生物技术研究所,北京100050

出　　处：《中国新药杂志》2023年第23期2417-2424,共8页Chinese Journal of New Drugs

摘　　要：目的:探讨自然杀伤细胞(NK细胞)对乳腺癌细胞的体外杀伤活性以及与低浓度拉帕替尼联合杀伤乳腺癌细胞的作用效果和可能的作用机制。方法:采用CCK-8方法检测不同浓度拉帕替尼对乳腺癌细胞24,48和72 h增殖活性的影响;乳酸脱氢酶(lactate dehydrogenase,LDH)检测试剂盒检测NK细胞以及NK细胞联合拉帕替尼对肿瘤细胞的杀伤作用;流式细胞术检测拉帕替尼作用前后人表皮生长因子受体2(HER-2)低表达乳腺癌细胞MDA-MB-231和BT549表面HER-2表达水平的变化;流式细胞术检测乳腺癌细胞表面NK细胞激活性和抑制性受体的配体表达情况。结果:拉帕替尼对乳腺癌细胞增殖活性的抑制与作用时间和浓度成正比,对HER-2高表达乳腺癌细胞的增殖抑制率要大于HER-2低表达乳腺癌细胞;NK细胞对乳腺癌细胞均具有一定的杀伤活性,拉帕替尼(HER-2高表达乳腺癌细胞MDA-MB-453和BT474选择拉帕替尼的终浓度为0.1μmol·L^(-1),HER-2低表达乳腺癌细胞MDA-MB-231和BT549选择拉帕替尼的终浓度为3.125μmol·L^(-1))联合NK细胞能显著提高NK细胞对乳腺癌细胞的杀伤效率;拉帕替尼(3.125μmol·L^(-1))与HER-2低表达乳腺癌细胞作用后能提高HER-2低表达乳腺癌细胞表面HER-2的表达水平。结论:拉帕替尼能够提高NK细胞对乳腺癌细胞的体外杀伤活性,其协同作用机制可能与拉帕替尼提高乳腺癌细胞表面HER-2的表达水平有关。Objective:To study the cytotoxicity effect of NK cells on breast cancer cells in vitro,and the cytotoxicity and possible mechanism of NK cells combined with low concentration lapatinib on breast cancer cells.Methods:CCK⁃8 method was used to detect the effect of different concentrations of lapatinib on the proliferation of breast cancer cells for 24 h,48 h and 72 h.LDH Assay Kit was applied to detect cytotoxicity of NK cells and NK cells combined with lapatinib on tumor cells.Changes of HER⁃2 expression on the surface of MDA⁃MB⁃231 and BT549 breast cancer cells before and after treatment with lapatinib and the ligand expression of NK cell activated and inhibitory receptors on breast cancer cell surface were both evaluated by flow cytometry.Results:The inhibitory effect of lapatinib on the proliferative activity of breast cancer cells was proportional to the time and concentration of action.The inhibitory rate on the proliferation of HER⁃2 high⁃expression breast cancer cells was greater than that of HER⁃2 low⁃expression breast cancer cells.NK cells had certain cytotoxicity against breast cancer cells,and lapatinib(The final concentration of lapatinib was 0.1μmol·L^(-1) in HER⁃2 high⁃expression breast cancer cells MDA⁃MB⁃453 and BT474 and 3.125μmol·L^(-1) in HER⁃2 low⁃expression breast cancer cells MDA⁃MB⁃231 and BT549)combined with NK cells could significantly improve the killing efficiency of NK cells against breast cancer cells.Lapatinib(3.125μmol·L^(-1))could increase the expression level of HER⁃2 on the surface of HER⁃2 low⁃expressionbreast cancer cells after interacting with HER⁃2 low⁃expression breast cancer cells.Conclusion:Lapatinibimproves the killing activity of NK cells against breast cancer cells in vitro,and its synergistic mechanism may berelated to the increased expression of HER⁃2 on the surface of breast cancer cells by lapatinib.

关 键 词：自然杀伤细胞 乳腺癌 拉帕替尼 免疫治疗 

分 类 号：R967[医药卫生—药理学]