雷公藤内酯醇联合顺铂对食管鳞癌细胞影响的机制研究

发布时间：2018-05-28 15:34

本文选题：食管鳞癌细胞 + 雷公藤内酯醇(Triptolide　；参考：《新乡医学院》2017年硕士论文

【摘要】：背景雷公藤内酯醇(Triptolide,TPL)是一种从植物雷公藤中提取的二萜内酯类化合物,具有免疫抑制、抗炎、抗增殖等多种生物学活性,是临床上公认的免疫抑制剂,主要用于治疗类风湿性关节炎、系统性红斑狼疮等疾病。近年来大量研究发现,TPL对乳腺癌、肺癌、肝癌、白血病等多种肿瘤均有不同程度的抑制作用。其机制与抑制肿瘤细胞增殖、促进肿瘤细胞凋亡、抑制肿瘤细胞的侵袭与转移等密切相关。而雷公藤内酯醇在抗食管癌等方面尚无相关研究。目的观察雷公藤内酯醇(TPL)及其联合顺铂对人食管鳞癌细胞株ECA-109体外活性的影响及可能机制,以期为治疗食管癌提供新的思路和理论依据。方法1.选取对数生长期的人食管鳞癌ECA-109细胞株,并进行以下分组:空白对照组(不经药物处理),单用TPL处理组(TPL浓度分别为6.25、12.5、25、50、100μg·L-1),单用顺铂处理组(浓度为4.17μmol·L-1),TPL联合顺铂组(TPL浓度为25μg·L-1,顺铂浓度为4.17μmol·L-1)。2.采用四甲基偶氮唑蓝法(MTT)测定各组不同时间段(24、48、72 h)的细胞增殖抑制率,并计算两药之间的交互作用。3.采用Western Blotting法检测各组细胞作用24h后活化型半胱氨酸天冬氨酸特异性蛋白酶-3(cleaved-Caspase-3)的表达情况。结果1.MTT结果显示:同一时间点,不同浓度TPL组对ECA-109细胞增殖的抑制率明显高于空白对照组(P0.05)且呈剂量依赖性(P0.05);在同一药物浓度条件下,随作用时间延长,细胞增殖抑制率明显增加(P0.05)且呈时间依赖性。同一时间点,TPL联合顺铂组对ECA-109细胞增殖的抑制率高于相同浓度各单药组(P0.05)。2.Western-blotting结果显示:各组细胞处理24 h后,单用TPL组、顺铂组及联合用药组cleaved-caspase-3蛋白表达量均显著高于空白对照组(P0.05),且联合两药用药组高于单用TPL、顺铂组(P0.05)。结论TPL具有抑制食管癌ECA-109细胞增殖的作用,TPL联合顺铂能协同增加对食管癌ECA-109细胞的抑制,其机制与cleaved-caspase-3蛋白表达上调有关。
[Abstract]:Background Triptolide triptolide (TPL) is a kind of diterpene lactones extracted from plant triptolide, which has many biological activities, such as immunosuppressive, anti-inflammatory, anti-proliferation and so on, so it is recognized as an immunosuppressant in clinic. Mainly used in the treatment of rheumatoid arthritis, systemic lupus erythematosus and other diseases. In recent years, it has been found that TPL can inhibit many kinds of tumors, such as breast cancer, lung cancer, liver cancer, leukemia and so on. The mechanism is closely related to the inhibition of tumor cell proliferation, the promotion of tumor cell apoptosis and the inhibition of invasion and metastasis of tumor cells. However, triptolide has not been studied on esophageal cancer. Objective to observe the effect of triptolide and its combination with cisplatin on the activity of human esophageal squamous cell carcinoma cell line ECA-109 in vitro and its possible mechanism in order to provide a new idea and theoretical basis for the treatment of esophageal carcinoma. Method 1. Human esophageal squamous cell carcinoma (ECA-109) cell lines with logarithmic growth period were selected. They were divided into the following groups: the concentration of TPL in the blank control group was 6.25 渭 g / L ~ (-1), that in the single TPL group was 6.25 渭 g / L ~ (-1) and that in the control group was 4.17 渭 mol / L ~ (-1) / L ~ (-1) and 4.17 渭 mol / L ~ (-1) / L ~ (-1) ~ (-1) ~ (-1) respectively, and that in the cisplatin group was 25 渭 g / L ~ (-1) and 4.17 渭 mol / L ~ (-1) 路L ~ (-1) 路L ~ (-1) respectively. The inhibition rate of cell proliferation was measured by MTT method in each group at different time points (24 48.72 h), and the interaction between the two drugs was calculated. The expression of activated cysteine aspartate specific protease-3 cleaved-Caspase-3 was detected by Western Blotting assay. Results 1.MTT showed that at the same time, the inhibition rate of ECA-109 cell proliferation in different concentrations of TPL was significantly higher than that in control group (P0.05) and in a dose-dependent manner. The inhibitory rate of cell proliferation was significantly increased in a time dependent manner (P 0.05). The inhibitory rate of TPL combined with cisplatin on the proliferation of ECA-109 cells at the same time point was higher than that in each single drug group at the same concentration. 2. The results of Western-blotting showed that the cells in each group were treated for 24 hours and treated with TPL alone. The expression of cleaved-caspase-3 protein in the cisplatin group and the combination group was significantly higher than that in the blank control group (P 0.05), and the expression of cleaved-caspase-3 protein in the combination group was higher than that in the single administration group and in the cisplatin group. Conclusion TPL can inhibit the proliferation of esophageal carcinoma ECA-109 cells. TPL combined with cisplatin can inhibit the proliferation of esophageal carcinoma ECA-109 cells. The mechanism is related to the up-regulation of cleaved-caspase-3 protein expression.
【学位授予单位】：新乡医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.1

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