EGCG单用/与CAR-T疗法联合应用对Raji细胞杀伤、THP-1细胞炎症因子mRNA表达抑制作用  

EGCG alone/combined with CAR-T therapy can kill Raji cells and inhibit the mRNA expression of THP-1 inflammatory factor

作　　者：史甲儒 姜淇耀 罗志强 于国华 王建勋 SHI Jiaru;JIANG Qiyao;LUO Zhiqiang;YU guohua;WANG Jianxun(School of Life Sciences,Beijing University of Chinese Medicine,Beijing 102488,China)

机构地区：[1]北京中医药大学生命科学学院,北京102488 [2]中国中医科学院中药资源中心道地药材国家重点实验室 [3]深圳北京中医药大学研究院细胞与基因治疗研究所

出　　处：《山东医药》2023年第24期1-6,共6页Shandong Medical Journal

基　　金：北京市双一流高层次人才科研启动经费项目(9011451310032)。

摘　　要：目的观察表没食子儿茶素没食子酸酯(EGCG)单用/与嵌合抗原受体T细胞(CAR-T)疗法联合应用对人Burkitt’s淋巴瘤细胞株Raji杀伤、人单核细胞白血病细胞系THP-1炎症因子mRNA表达抑制作用。方法取对数生长期的Raji细胞,并分为4组,对照组为单纯Raji细胞,Raji+CAR-T组加入CAR-T,Raji+EGCG组加入EGCG,Raji+CAR-T+EGCG组加入CAR-T和EGCG,荧光素酶化学发光法和Annexin V-FITC染色法检测各组细胞相对活力及杀伤效率;取对数生长期的人单核细胞白血病细胞(THP-1细胞),并分为对照组、模型组、EGCG组,其中模型组和EGCG组加入LPS(终浓度1μg/mL)诱导炎症模型,同时EGCG组加入EGCG(浓度为100μmol/mL)干预,采用实时荧光定量PCR法检测COX-2、IL-6、IL-8、TNF-αmRNA相对表达量。结果Raji+CAR-T+EGCG组、Raji+EGCG组、Raji+CAR-T组、对照组细胞相对活力分别为52.93±4.61、91.67±6.79、51.26±2.61、100.00±5.45,对照组与其余3组比较,Raji+CAR-T+EGCG组、Raji+EGCG组比较,P均<0.05。Raji+CAR-T组、Raji+EGCG组、Raji+CAR-T+EGCG组的杀伤效率分别为10.29%、16.21%、16.65%,对照组与其余3组比较,Raji+CAR-T组、Raji+CAR-T+EGCG组比较,P均<0.05。EGCG组COX-2、IL-6、IL-8、TNF-αmRNA相对表达量分别为1.49±0.20、2.09±0.91、5.25±0.36、1.07±0.42,模型组分别为7.06±0.72、34.25±9.98、6.97±0.58、6.64±1.71,对照组分别为1、1、1、1,模型组与其余两组比较,P均<0.05。结论EGCG不仅可以提升CAR-T活力,并减缓其耗竭,同时在不影响CAR-T杀伤能力和效率的前提下,抑制Raji细胞增殖及多种炎症因子表达。Objective To observe the killing effect of epigallocatechin gallate(EGCG)alone/combined with chimeric antigen receptor T cell(CAR-T)therapy on human Burkitt′s lymphoma cells Raji and its inhibitory effect on the mRNA expression of THP-1 inflammatory factor in human monocytic leukemia cell line.Methods Raji cells in the logarithmic growth phase were selected and divided into four groups.Cells in the control group were pure Raji cells,cells in the Raji+CAR-T group were added with CAR-T,the Raji+EGCG group with EGCG,and the Raji+CAR-T+EGCG group with CAR-T and EGCG,respectively.The relative viability and killing efficiency of cells in each group were detected by luciferase chemiluminescence and Annexin V-FITC staining.Human monocytic leukemia cells(THP-1 cells)in the logarithmic growth phase were selected and divided into the control group,model group and EGCG group.LPS(with final concentration of 1μg/mL)was added to the model group and EGCG group to induce inflammation models,while cells in the EGCG group were added with EGCG(100μmol/mL).Real-time fluorescent quantitative PCR was used to detect the relative expression levels of cyclooxygenase-2(Cox-2),interleukin-6(Il-6),interleukin-8(Il-8),tumor necrosis factor-α(TNF-α)mrna.Results The relative cell activity of the Raji+CAR-T+EGCG group,Raji+EGCG group,Raji+CAR-T group,and control group was 52.93±4.61,91.67±6.79,51.26±2.61,and 100.00±5.45,respectively.Significant difference was found between the Raji+CAR-T+EGCG group and Raji+EGCG group,as well as between the control group and the rest of three groups(all P<0.05).The killing efficiencies of the Raji+CAR-T group,Raji+EGCG group,and Raji+CAR-T+EGCG group were 10.29%,16.21%and 16.65%,respectively.Significant difference was found between the control group and the other three groups as well as between the Raji+CAR-T+EGCG group and Raji+CAR-T+EGCG group(all P<0.05).The relative expression levels of COX-2,IL-6,IL-8 and TNF-αmRNA in the EGCG group were 1.49±0.20,2.09±0.91,5.25±0.36,and 1.07±0.42,respectivel

关 键 词：表没食子儿茶素没食子酸酯 嵌合抗原受体T细胞疗法 Burkitt’s淋巴瘤 RAJI细胞 

分 类 号：R285.5[医药卫生—中药学]