發(fā)布時間：2018-05-07 13:28

  本文選題：順鉑 + 小豆蔻明　； 參考：《福建醫(yī)科大學》2014年碩士論文

【摘要】：目的 鉑類藥物化療為卵巢癌治療的重要方法，，但化療過程中腫瘤細胞耐藥性的產(chǎn)生嚴重影響藥物的療效。目前，采用其它抗腫瘤藥物與鉑類聯(lián)用而達到治療效果是臨床常用的治療方案。因此，尋找能提高鉑類治療卵巢癌療效的藥物臨床意義重大。小豆蔻明（Cardamonin, CAR）為姜科植物草豆蔻中提取的黃酮類單體。國內(nèi)外的研究表明，CAR具有抗腫瘤、抗炎等多種藥理活性，能增強TRAIL的抗腫瘤作用，其作用機制與增強死亡受體以及凋亡蛋白的表達有關(guān)。然而，CAR能否增強順鉑（cisplatin, Cis）的腫瘤抑制作用尚不明確。本課題旨在考察CAR聯(lián)合Cis對卵巢癌SKOV3細胞增殖的作用，并探明其作用機制，為聯(lián)合鉑類抗腫瘤化療藥物的研發(fā)提供新方向，為CAR進一步開發(fā)奠定理論基礎(chǔ)及提供實驗依據(jù)。 方法 1分組與給藥 SKOV3分為正常對照組（control），溶劑對照組（0.1%DMSO），Cis組（2μg/mL）、CAR組（20μM）和Cis+CAR組（2μg/mL+20μM）。 2測定指標及方法 2.1MTT法和克隆形成實驗測定細胞增殖； 2.2PI染色法檢測細胞周期； 2.3DNA碎片法檢測細胞凋亡； 2.4MDC染色法檢測細胞自噬； 2.5RT-PCR法檢測谷胱甘肽-S-轉(zhuǎn)移酶-π （glutathione S-transferase-π, GST-π)、多藥耐藥蛋白（multidrug resistance protein, MRP）的mRNA表達； 2.6Western Blot法檢測Bcl-2、XIAP、survivin和LC3的蛋白表達。 結(jié)果 1CAR聯(lián)合Cis用藥能夠顯著性抑制SKOV3細胞增殖，其抑制率明顯高于Cis單獨用藥（P 0.01），20μM CAR與2μg/mL Cis具有協(xié)同作用； 2CAR聯(lián)合Cis對SKOV3細胞克隆形成的抑制顯著強于單獨用藥（P 0.01）； 3與單獨用藥相比，CAR聯(lián)合Cis對DNA碎片的生成無顯著性差異； 4Cis能夠誘導GST-π mRNA表達的增加（P 0.01），而對MRP的表達無明顯影響；CAR能夠降低MRP mRNA的表達（P 0.01），而對GST-π的表達無明顯影響；兩藥聯(lián)合時，CAR能夠降低Cis誘導的GST-π高表達，同時降低MRP的表達（P 0.01）； 5Cis和CAR單獨處理后，XIAP與survivin的表達降低（P 0.01）；而聯(lián)合給藥組Bcl-2、XIAP、survivin的表達顯著性降低，其抑制作用明顯高于單獨給藥組（P 0.01）。 6Cis和CAR處理過的細胞內(nèi)MDC染色顆粒數(shù)增加；而Cis和CAR聯(lián)用后，細胞內(nèi)MDC染色顆粒數(shù)目顯著性增加，二者具有協(xié)同誘導自噬的作用。 結(jié)論 1CAR與Cis具有協(xié)同抑制卵巢癌SKOV3細胞增殖的作用； 2CAR聯(lián)合Cis具有增強卵巢癌SKOV3細胞自噬，抑制細胞周期的作用； 3CAR增強Cis抗腫瘤的作用與抗凋亡蛋白和耐藥基因表達的降低密切相關(guān)。
[Abstract]:Purpose Chemotherapy with platinum drugs is an important method in the treatment of ovarian cancer, but the drug resistance of tumor cells in the course of chemotherapy seriously affects the efficacy of the drugs. At present, the use of other antitumor drugs combined with platinum to achieve therapeutic effect is commonly used in clinical treatment. Therefore, it is of great clinical significance to find drugs that can improve the efficacy of platinum in the treatment of ovarian cancer. Cardamonin (Carr) is a flavonoid monomer extracted from Curcumaceae. Studies at home and abroad have shown that car has many pharmacological activities, such as anti-tumor, anti-inflammatory and so on, which can enhance the anti-tumor effect of TRAIL, and its mechanism is related to the enhancement of the expression of death receptor and apoptotic protein. However, whether car can enhance the tumor inhibitory effect of cisplatin (Cis) is not clear. The purpose of this study was to investigate the effect of CAR combined with Cis on the proliferation of ovarian cancer SKOV3 cells, and to explore its mechanism, to provide a new direction for the research and development of combined platinum antitumor chemotherapeutic agents, and to lay a theoretical foundation and experimental basis for the further development of CAR. Method 1 grouping and administration SKOV3 was divided into normal control group (n = 20), solvent control group (n = 0.1) and Cis CAR group (n = 2 渭 g / mL) and Cis CAR group (n = 2 渭 g/mL = 20 渭 M). 2 determination index and method Cell proliferation was determined by 2.1MTT assay and clone formation assay. Cell cycle was detected by 2.2PI staining. Apoptosis was detected by 2.3DNA fragment assay. Autophagy was detected by 2.4MDC staining. The mRNA expression of glutathione S-transferase- 蟺, glutathione S-transferase- 蟺 and multidrug resistance protein, MRP) was detected by 2.5RT-PCR. The expression of survivin and LC3 were detected by 2.6Western Blot. Result 1CAR combined with Cis could significantly inhibit the proliferation of SKOV3 cells, and the inhibitory rate was significantly higher than that of 20 渭 M CAR and 2 渭 g/mL Cis of Cis alone. The inhibitory effect of 2CAR combined with Cis on SKOV3 cell clone formation was significantly stronger than that of single drug (P 0.01). (3) there was no significant difference in the formation of DNA fragments between car and Cis alone. 4Cis could induce the increase of mRNA expression of GST- 蟺, but had no effect on the expression of MRP. Car could decrease the expression of GST- 蟺, but not the expression of GST- 蟺, and the combination of two drugs could decrease the overexpression of GST- 蟺 induced by Cis. At the same time, the expression of MRP was decreased (P 0.01). The expression of survivin and XIAP decreased significantly in 5Cis and CAR alone, but the expression of Bcl-2TXIAPP in combination group was significantly lower than that in control group (P 0.01). The number of MDC staining granules treated with 6Cis and CAR increased, while that of Cis and CAR increased significantly, both of them could induce autophagy synergistically. Conclusion 1CAR and Cis could inhibit the proliferation of ovarian cancer SKOV3 cells. 2CAR combined with Cis can enhance autophagy of SKOV3 cells and inhibit cell cycle. The role of 3CAR in enhancing the anti-tumor effect of Cis is closely related to the decrease of anti-apoptotic protein and drug resistance gene expression.
【學位授予單位】：福建醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R737.31


本文編號：1857104