摘要
目的评价红色诺卡菌细胞壁骨架(Nocardia rubracell wall skeleton,Nr-CWS)多糖组分对宫颈癌HeLa细胞增殖的抑制作用,对巨噬细胞、NK、LAK、CTL细胞增殖活化的诱导以及对HeLa细胞的杀伤作用,并阐述其诱导免疫发挥抗肿瘤作用的机制。方法 MTT法检测Nr-CWS多糖组分对HeLa细胞的抑制作用和促进NK、LAK、CTL细胞及对宫颈癌HeLa细胞的杀伤作用;采用ELISA法分别测定经Nr-CWS多糖组分作用后NK、LAK、CTL细胞中IL-12、IFN-γ等相关炎症因子水平,Real-time PCR检测NK、LAK和CTL细胞NKG2D mRNA表达,Western blot检测NK、LAK和CTL细胞NKG2D蛋白表达和HeLa细胞NKG2D配体ULBP1、ULBP2和MICA的蛋白表达;评价Nr-CWS多糖组分诱导免疫和杀伤肿瘤细胞的作用。结果 Nr-CWS多糖提取物具有明显增强NK、LAK、CTL细胞增殖活化和杀伤宫颈癌HeLa细胞的作用,并呈剂量-时间依赖性,同时上调HeLa细胞NKG2D配体表达,使其敏感性增强。结论 Nr-CWS多糖提取物可通过诱导NK、LAK、CTL细胞增殖活化,增强其对宫颈癌HeLa细胞的杀伤作用,同时使HeLa细胞对前者敏感性增强,从而发挥抗肿瘤作用。
This study was designed to evaluate the inhibitory effect of Nocardia rubra cell wall skeleton(NrCWS) polysaccharide components on the proliferation of cervical cancer HeLa cells, and the inducing effects on the proliferation and activation of macrophages, NK, LAK, CTL cells, thus to elucidate the mechanism of inducing immunity to play an anti-tumor role. MTT was used to evaluate the inhibitory effect of Nr-CWS polysaccharide components on HeLa cells and the killing effects of NK, LAK, CTL cells on HeLa cells;ELISA method was used to detect the level of IL-12, IFN-γ and other related inflammatory factors in NK, LAK, and CTL cells after the treatment of Nr-CWS polysaccharide component. Real-time PCR was used to detect the NKG2 D mRNA level in NK, LAK, CTL cells after Nr-CWS treatment;Western blot was used to detect NKG2 D protein expression in NK,LAK, CTL cells, and the protein expressions of NKG2 D ligands including ULBP1, ULBP2, and MICA in HeLa cells.Data showed that Nr-CWS polysaccharide extract significantly enhanced the proliferation and activation of NK,LAK, CTL cells and killed HeLa cells in a dose-time manner, and also up-regulated the expression of NKG2 D ligand in HeLa cells to enhance its sensitivity. Therefore, we speculate Nr-CWS polysaccharide extract can enhance the anti-tumor effect of NK, LAK and CTL cells by inducing their proliferation and activation and enhancing the sensitivity of HeLa cells.
作者
王筱婧
于丹
王菲
陈风华
李伟
张海
WANG Xiaojing;YU Dan;WANG Fei;CHEN Fenghua;LI Wei;ZHANG Hai(Chinese Pharmaceutical Association,Beijing 100050,China;School of Pharmacy,Shandong University of Traditional Chinese Medicine,Ji’nan 250355,China;Pharmaceutical Preparation Section,Shanghai First Maternity and Infant Hospital,Tongji University,Shanghai 201204,China)
出处
《免疫学杂志》
CAS
CSCD
北大核心
2020年第12期1039-1045,共7页
Immunological Journal
基金
上海市卫健委科研面上项目(201740217)。
