摘要
目的:验证中药有效成分大黄酸抑制低氧引起的结肠癌细胞中免疫抑制分子的表达,为大黄酸与效应T细胞的联合应用提供基础理论依据。方法:首先根据CCK-8检测大黄酸对结肠癌细胞系的细胞毒作用,选择恰当的研究浓度;然后用Western blot和q PCR检测在低氧条件下,一定浓度的大黄酸能否下调缺氧诱导因子-1α(HIF-1α)和某些免疫抑制分子的水平;最后在我们自己建立的能够被T淋巴细胞特异识别杀伤的结肠癌细胞模型上,进一步检测大黄酸在低氧条件下对效应T细胞活性的影响。结果:与常氧对照组相比,低氧组、低氧+25μM大黄酸组和低氧+50μM大黄酸组细胞中程序性死亡受体-配体1(PD-L1)m RNA的相对量分别为1.63±0.29、0.80±0.17和0.47±0.12;半乳糖凝集素-1(galectin-1)m RNA的相对量分别为4.42±0.73、0.47±0.11和0.24±0.11。当效靶比为1:1、2:1和4:1时,T细胞对肿瘤细胞的杀伤百分率分别为(27.32±3.05)%、(42.68±3.69)%和(54.02±1.82)%;而低氧条件下,杀伤百分率分别下降至(21.53±3.74)%、(33.55±1.51)%和(45.20±2.27)%;在低氧条件下加入50μM大黄酸,杀伤百分率又分别升高至(32.70±2.37)%、(47.01±3.15)%和(57.26±1.12)%,各组之间有显著性差异(P<0.05)。结论:一定浓度的大黄酸能够降低低氧引起的免疫抑制分子表达升高,并且可以逆转由于低氧产生的效应T细胞活性下降。大黄酸有望与T细胞过继免疫疗法联合使用,用于结肠癌的治疗。
Objecive To study the abilities of rhein in inhibiting the increase of immunosuppressive molecules in colorectal cancer cells caused by hypoxia and provide a theoretical basis for the combined application of rhein and effector lymphocytes.Method CCK-8 assay was performed to choose optional concentration of rhein,Western blot and qPCR were used to detect the level of HIF-1α(hypoxia-inducible factor-1α)and immunosuppressive molecules such as PD-L1(programmed cell death ligand-1)and galectin-1 in colorectal cancer cells treated by rhein and hypoxic condition,and the cytotoxity of effect T lymphocyte with rhein and hypoxia was explored on the colorectal cancer cell model which was recognized and killed specifically by effect T lymphocyte.Result Comparing to the normoxia group,the relative quantities of PDL1 mRNA of hypoxia group,hypoxia+25μM rhein group and hypoxia+50μM rhein group were 1.63±0.29,0.80±0.17 and 0.47±0.12,and the relative quantities of galectin-1 mRNA were 4.42±0.73,0.47±0.11 and 0.24±0.11 respectively.In normoxia,at the indicated effector:target ratio(1:1,2:1 and 4:1),the percents of cell lysis were(27.32±3.05)%,(42.68±3.69)%and(54.02±1.82)%;but in hypoxia,they were decreased to(21.53±3.74)%,(33.55±1.51)%and(45.20±2.27)%;when 50μM rhein was added,they were recovered to(32.70±2.37)%,(47.01±3.15)%and(57.26±1.12)%.The differences were significant between the hypoxia group and normoxia group or hypoxia+rhein group(P<0.05).Conclusion Rhein could decrease the levels of immunosuppressive molecules of colorectal cancer cells and enhance the cytotoxity of effect T lymphocyte to colorectal cancer cells in hypoxia.Rhein had great potential to combine with effect T lymphocyte for treatment of colorectal cancer.
作者
袁向飞
田文聪
谢俊木子
王丰
YUAN Xiang-fei;TIAN Wen-cong;XIE Jun-mu-zi(Tianjin Institute of Integrative Medicine for Acute Abdominal Diseases,Tianjin Nankai Hospital,Tianjin(300100),China)
出处
《中国中西医结合外科杂志》
CAS
2018年第4期476-481,共6页
Chinese Journal of Surgery of Integrated Traditional and Western Medicine
基金
国家自然科学基金面上项目(81572318)
关键词
大黄酸
效应T细胞
免疫抑制分子
低氧
结肠癌
Rhein
effect T lymphocyte
immunosuppressive molecule
hypoxia
colorectal cancer
