摘要
目的探讨青蒿素衍生物双氢青蒿素(DHA)对非小细胞肺癌(NSCLC)细胞诱导的CD8^(+)T细胞抗肿瘤免疫功能的调控作用。方法将NSCLC细胞系A549细胞分为二甲基亚砜(DMSO)对照组和DHA处理组;分别用DMSO和不同浓度的DHA(25、50和100μmol/L)处理A549细胞,根据半抑制浓度(IC 50)选择最适浓度的DHA处理A549细胞0、24、48、72 h;CCK-8法和集落形成实验检测DHA对A549细胞增殖和集落形成能力的影响;密度梯度离心法分离健康个体外周血单个核细胞(PBMC),经黏附贴壁法去除单核细胞,随后与丝裂霉素C预处理的A549细胞按照10∶1比例共培养,2周后采用流式细胞术分别检测CD8^(+)T细胞比例及其穿孔素和颗粒酶B的表达水平。结果与对照组相比,25、50和100μmol/L DHA处理24 h后的A549细胞增殖的抑制率均升高(P<0.01);DHA对A549细胞的IC 50为46.26μmol/L;依据IC 50浓度检测50μmol/L DHA处理A549细胞0、24、48、72 h的细胞抑制率分别为1.53%、53.50%、63.84%和69.91%,分别与前一观察时间点即0、24和48 h相比,细胞抑制率均增高(P<0.01);集落形成实验结果显示,与对照组相比,DHA处理组的A549细胞集落形成数减少(P<0.01);流式细胞术结果显示,与对照组相比,DHA预处理组的A549细胞在共培养体系中诱导的CD8^(+)T细胞的比例、表达穿孔素和颗粒酶B的CD8^(+)T细胞比例均更高(P<0.01)。结论DHA能够抑制NSCLC细胞生长,促进NSCLC细胞诱导的CD8^(+)T细胞抗肿瘤免疫应答。
Objective To investigate the regulatory effect of artemisinin derivative dihydroartemisinin(DHA)on anti-tumor immune function of CD8^(+)T cells induced by non-small cell lung cancer(NSCLC)cells.Methods NSCLC A549 cells were divided into DMSO control group and DHA treatment group.A549 cells were treated with DMSO and DHA at different concentrations(25,50 and 100μmol/L),and the optimal concentration of DHA was selected to treat A549 cells for 0,24,48 and 72 h according to half maximal inhibitory concentrate(IC 50).CCK-8 method and colony formation test were used to detect the effect of DHA on the proliferation and colony formation ability of A549 cells.Peripheral blood mononuclear cells(PBMCs)of healthy individuals were isolated by density gradient centrifugation.After monocytes were removed by adhesion method,A549 cells pretreated with mitomycin C were co-cultured with PBMCs at 10:1 ratio.After 2 weeks,flow cytometry was used to detect the proportion of CD8^(+)T cells and the expression levels of perforin and granzyme B.Results Compared with the control group,the proliferation inhibition rates of A549 cells increased after treatment with 25,50 and 100μmol/L DHA for 24 h(P<0.01).The IC 50 of DHA on A549 cells was 46.26μmol/L.According to IC 50 concentration analysis,the inhibition rates of A549 cells treated with 50μmol/L DHA for 0,24,48 and 72h were 1.53%,53.50%,63.84%and 69.91%,and the cells inhibition rates of A548 cells increased compared with the previous observation time point,namely 0,24 and 48 h(P<0.01).The colony formation assay showed that the colony formation number of A549 cells in DHA treated group decreased compared with the control group(P<0.01).Flow cytometry results showed that compared with the control group,the proportion of CD8^(+)T cells induced by A549 cells in the co-culture system and the proportion of CD8^(+)T cells expressing perforin and granzyme B were higher in DHA pretreatment group(P<0.01).Conclusion DHA inhibits the growth of NSCLC cells and promotes anti-tumor immune response of CD8^(+)T cells induced by NSCLC cells.
作者
王南楠
刘宇
凌惠娟
牛可
朱亚玉
陈礼文
Wang Nannan;Liu Yu;Ling Huijuan;Niu Ke;Zhu Yayu;Chen Liwen(Dept of Clinical Laboratory,The Second Affiliated Hospital of Anhui Medical University,Hefei 230601;Dept of Blood Transfusion,The Second Affiliated Hospital of Anhui Medical University,Hefei 230601)
出处
《安徽医科大学学报》
CAS
北大核心
2024年第3期424-429,共6页
Acta Universitatis Medicinalis Anhui
基金
安徽省高校自然科学研究重点项目(编号:2023AH053170)
安徽省临床医学研究转化专项项目(编号:202304295107020019)。
