本文選題：食管鱗癌細(xì)胞 + 雷公藤內(nèi)酯醇(Triptolide��； 參考：《新鄉(xiāng)醫(yī)學(xué)院》2017年碩士論文

【摘要】：背景雷公藤內(nèi)酯醇(Triptolide,TPL)是一種從植物雷公藤中提取的二萜內(nèi)酯類化合物,具有免疫抑制、抗炎、抗增殖等多種生物學(xué)活性,是臨床上公認(rèn)的免疫抑制劑,主要用于治療類風(fēng)濕性關(guān)節(jié)炎、系統(tǒng)性紅斑狼瘡等疾病。近年來大量研究發(fā)現(xiàn),TPL對乳腺癌、肺癌、肝癌、白血病等多種腫瘤均有不同程度的抑制作用。其機(jī)制與抑制腫瘤細(xì)胞增殖、促進(jìn)腫瘤細(xì)胞凋亡、抑制腫瘤細(xì)胞的侵襲與轉(zhuǎn)移等密切相關(guān)。而雷公藤內(nèi)酯醇在抗食管癌等方面尚無相關(guān)研究。目的觀察雷公藤內(nèi)酯醇(TPL)及其聯(lián)合順鉑對人食管鱗癌細(xì)胞株ECA-109體外活性的影響及可能機(jī)制,以期為治療食管癌提供新的思路和理論依據(jù)。方法1.選取對數(shù)生長期的人食管鱗癌ECA-109細(xì)胞株,并進(jìn)行以下分組:空白對照組(不經(jīng)藥物處理),單用TPL處理組(TPL濃度分別為6.25、12.5、25、50、100μg·L-1),單用順鉑處理組(濃度為4.17μmol·L-1),TPL聯(lián)合順鉑組(TPL濃度為25μg·L-1,順鉑濃度為4.17μmol·L-1)。2.采用四甲基偶氮唑藍(lán)法(MTT)測定各組不同時間段(24、48、72 h)的細(xì)胞增殖抑制率,并計算兩藥之間的交互作用。3.采用Western Blotting法檢測各組細(xì)胞作用24h后活化型半胱氨酸天冬氨酸特異性蛋白酶-3(cleaved-Caspase-3)的表達(dá)情況。結(jié)果1.MTT結(jié)果顯示:同一時間點(diǎn),不同濃度TPL組對ECA-109細(xì)胞增殖的抑制率明顯高于空白對照組(P0.05)且呈劑量依賴性(P0.05);在同一藥物濃度條件下,隨作用時間延長,細(xì)胞增殖抑制率明顯增加(P0.05)且呈時間依賴性。同一時間點(diǎn),TPL聯(lián)合順鉑組對ECA-109細(xì)胞增殖的抑制率高于相同濃度各單藥組(P0.05)。2.Western-blotting結(jié)果顯示:各組細(xì)胞處理24 h后,單用TPL組、順鉑組及聯(lián)合用藥組cleaved-caspase-3蛋白表達(dá)量均顯著高于空白對照組(P0.05),且聯(lián)合兩藥用藥組高于單用TPL、順鉑組(P0.05)。結(jié)論TPL具有抑制食管癌ECA-109細(xì)胞增殖的作用,TPL聯(lián)合順鉑能協(xié)同增加對食管癌ECA-109細(xì)胞的抑制,其機(jī)制與cleaved-caspase-3蛋白表達(dá)上調(diào)有關(guān)。
[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.
【學(xué)位授予單位】：新鄉(xiāng)醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R735.1

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