植物凝集素、唑来膦酸及乳康饮扩增γδT细胞对三阴性乳腺癌细胞的杀伤

Observation on the killing of triple negative breast cancer cells byγδT cells amplified by phytohemagglutinin,zoledronic acid and Rukangyin

目的观察乳康饮(Ru Kang Yin,RKY)刺激γδT细胞增殖对乳腺癌细胞MDA-MB-231的杀伤作用,为中医药治疗乳腺癌提供依据。方法实验分为以下4组,Ⅰ组:外周血单个核细胞(PBMC)对照组;Ⅱ组:PBMC+植物凝集素(PHA)组;Ⅲ组:PBMC+唑来膦酸(ZOL)组;Ⅳ组:PBMC+RKY组。在各组培养0 d和14 d时,用流式细胞术检测各组γδT细胞占PBMC的百分比。将用免疫磁珠分选的γδT细胞与荧光染料羧基荧光素二醋酸盐琥珀酰亚胺酯(CFSE)标记的乳腺癌MDA-MB-231细胞以10∶1的比例共培养,测定γδT细胞对乳腺癌MDA-MB-231细胞的杀伤力。结果培养0 d时,Ⅰ组~Ⅳ组γδT细胞占PBMC百分比分别为(3.81±0.27)%、(4.19±0.41)%、(3.94±0.13)%、(4.16±0.11)%,各组间差异无统计学意义(F=1.462,P=0.296)。培养14 d时,Ⅰ组~Ⅳ组γδT细胞占PBMC百分比分别为(4.70±0.29)%、(31.09±1.95)%、(25.91±3.77)%、(28.84±2.54)%,各组间差异有统计学意义(F=71.985,P=0.000)。Ⅰ组~Ⅳ组对MDA-MB-231细胞的杀伤率分别为(1.17±0.86)%、(1.56±0.13)%、(1.47±0.09)%、(2.01±0.16)%,各组间差异有统计学意义(F=24.649,P=0.000)。结论RKY可以刺激γδT细胞增殖,提高对MDA-MB-231细胞的杀伤力,为乳腺癌的中医药治疗提供实验依据。

Objective To observe the killing effect ofγδT cell proliferation stimulated by Rukangyin(Ru Kang Yin,RKY)on breast cancer cell line MDA-MB-231,so as to provide basis for the treatment of breast cancer with traditional Chinese medicine.Methods The experiment was divided into four groups:groupⅠ(control),peripheral blood mononuclear cells(PBMCs);groupⅡ,PBMCs treated with phytohemagglutinin(PHA);groupⅢ,PBMCs treated with zoledronic acid(ZOL);and groupⅣ,PBMCs treated with RKY.After 0 and 14 days of culture,the percentage ofγδT cells in PBMCs for each group was detected by flow cytometry.TheγδT-positive cells sorted by immunomagnetic beads and breast cancer MDA-MB-231 cells labeled with fluorescent dye carboxyfluorescein diacetate succinimidyl ester(CFSE)were co-cultured at 10∶1 to determine the cytotoxicity ofγδT cells to breast cancer MDA-MB-231 cells.Results At 0 day of culture,the percentages ofγδT cells in PBMCs in groupsⅠ-Ⅳwere(3.81±0.27)%,(4.19±0.41)%,(3.94±0.13)%,and(4.16±0.11)%,respectively,and there was no statistical difference among different groups(F=1.462,P=0.296).After 14 days of culture,the percentages ofγδT cells in PBMCs in groupsⅠ-Ⅳwere(4.70±0.29)%,(31.09±1.95)%,(25.91±3.77)%,and(28.84±2.54)%,respectively,and there was significant difference among different groups(F=71.985,P=0.000).The killing rates of MDA-MB-231 cells in groupsⅠ-Ⅳwere(1.17±0.86)%,(1.56±0.13)%,(1.47±0.09)%,and(2.01±0.16)%,respectively,and there was significant difference among different groups(F=24.649,P=0.000).Conclusion RKY can stimulate the proliferation ofγδT cells and improve their cytotoxicity to MDA-MB-231 cells,which provides a basis for the treatment of breast cancer with traditional Chinese medicine.