【摘要】：目的：體外研究表皮生長(zhǎng)因子受體（Epidermal growth factor receptor，EGFR）抑制劑吉非替尼（Gefitinib）和雷帕霉素靶蛋白（Mammalian target of rapamycin，mTOR）抑制劑依維莫司（Everolimus）在gefitinib耐藥鼻咽癌細(xì)胞中聯(lián)合作用的效果及機(jī)制。 方法：MTT法測(cè)定單藥及聯(lián)合用藥對(duì)人鼻咽癌細(xì)胞株HONE1的增殖抑制；流式細(xì)胞術(shù)檢測(cè)藥物單獨(dú)或聯(lián)合作用于HONE1細(xì)胞株48h、72h后，各組細(xì)胞凋亡及周期分布的改變；Western Blot檢測(cè)單藥及聯(lián)合用藥48h后，HONE1細(xì)胞中磷酸化AKT（Phospho-AKT，p-AKT）和磷酸化S6K（Phospho-S6K，p-S6K）表達(dá)水平的變化。 結(jié)果：MTT法結(jié)果顯示gefitinib和everolimus對(duì)HONE1細(xì)胞的生長(zhǎng)抑制均呈劑量依賴性，并由此計(jì)算出，gefitinib的IC50為17.92μmol/L，everolimus的IC50為2.46nmol/L，但二者的聯(lián)合作用并沒有表現(xiàn)出明顯的協(xié)同效應(yīng)（P0.05）；流式細(xì)胞術(shù)顯示gefitinib及everolimus作用于HONE1細(xì)胞時(shí)，可誘導(dǎo)凋亡，引起G0/G1期細(xì)胞阻滯，，兩種作用均具有時(shí)間依賴性，而二者聯(lián)合的效果并未表現(xiàn)出明顯的優(yōu)勢(shì)(P0.05)；Western Blot顯示gefitinib對(duì)p-AKT抑制作用不明顯，everolimus可下調(diào)p-S6K的表達(dá)，但同時(shí)上調(diào)了p-AKT的表達(dá)，聯(lián)合用藥對(duì)p-S6K和p-AKT表達(dá)的抑制作用并不明顯強(qiáng)于單藥。 結(jié)論：mTOR抑制劑everolimus并不能逆轉(zhuǎn)gefitinib耐藥鼻咽癌細(xì)胞株HONE1的耐藥性，兩者聯(lián)合的效果在鼻咽癌細(xì)胞中沒有明顯優(yōu)勢(shì)，EGFR/AKT信號(hào)通路與mTOR信號(hào)通路的關(guān)系及其在鼻咽癌細(xì)胞中的作用機(jī)制有待進(jìn)一步研究。
[Abstract]:Aim: to investigate the combined effect and mechanism of epidermal growth factor receptor (Epidermal growth factor receptor,EGFR) inhibitor gefitinib (Gefitinib) and rapamycin target protein (Mammalian target of rapamycin,mTOR) inhibitor (Evimostr (Everolimus) in gefitinib resistant nasopharyngeal carcinoma (NPC) cells in vitro. Methods the proliferation of human nasopharyngeal carcinoma (NPC) cell line HONE1 was inhibited by single drug and combined drug, and apoptosis and cell cycle distribution were detected by flow cytometry after treated with drugs alone or in combination for 48 h or 72 h. The expression of phosphorylated AKT (Phospho-AKT,p-AKT) and phosphorylated S6K (Phospho-S6K,p-S6K) in HONE1 cells were detected by Western Blot. Results the results of gefitinib and everolimus showed that the growth inhibition of HONE1 cells was dose-dependent, and the IC50 of gefitinib was 17.92 渭 mol/L,everolimus and the IC50 of HONE1 was 2.46 nmol / L, but the combined effect of the two showed no significant synergistic effect (P0.05). Flow cytometry showed that gefitinib and everolimus could induce apoptosis and induce cell arrest in G0/G1 phase when HONE1 cells were treated with gefitinib and everolimus. Both of the two effects were time-dependent, but the combined effect did not show obvious advantage (P0.05). Western Blot showed that the inhibitory effect of gefitinib on p-AKT was not obvious. Gefitinib could down-regulate the expression of p-S6K, but at the same time up-regulated the expression of p-AKT. The inhibitory effect of combined treatment on p-S6K and p-AKT expression was not significantly stronger than that of single drug. Conclusion everolimus can not reverse the drug resistance of gefitinib resistant nasopharyngeal carcinoma cell line HONE1. The relationship between EGFR / AKT signaling pathway and mTOR signaling pathway and its mechanism in nasopharyngeal carcinoma cells need to be further studied.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R739.63

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 徐錫金,楊海偉,霍霞;抑癌基因PTEN在鼻咽癌組織中的表達(dá)及意義[J];臨床耳鼻咽喉科雜志;2004年11期

2 ;Effects of cyclooxygenase 2 inhibitors on biological traits of nasopharyngeal carcinoma cells[J];Acta Pharmacologica Sinica;2004年07期

本文編號(hào)：2230990