本文選題：鼻咽癌 + 表皮生長因子受體��； 參考：《廣西醫(yī)科大學(xué)》2012年碩士論文

【摘要】：目的鼻咽癌的發(fā)生和發(fā)展是由多種病因通過多種途徑長期共同影響及誘導(dǎo)的過程,病因?qū)W的研究表明與EB病毒感染、地域環(huán)境因素影響、飲食習(xí)慣以及家族遺傳等因數(shù)相關(guān)。除此之外,某些特定蛋白和基因的過度表達及變異也在鼻咽癌發(fā)展中也發(fā)揮著重要作用。表皮生長因子受體(epidermal growth factor receptor, EGFR)在鼻咽癌(nasopharyngeal carcinoma, NPC)細胞中存在大量表達,并和鼻咽癌的發(fā)生和發(fā)展有著密切關(guān)系。本研究通過對比單純使用吉非替尼、單純照射及吉非替尼聯(lián)合照射對鼻咽細胞殺傷的體外實驗,旨在研究表皮生長因子受體的表達對放射照射的影響。探討吉非替尼對鼻咽癌放射治療作用的影響及可能機制,為臨床治療提供依據(jù)。方法取對數(shù)生長期人鼻咽癌CNE1和HONE1細胞接種于96孔板中并隨機分組,分組分別設(shè)立對照組、吉非替尼組、單純照射組及藥物照射組。吉非替尼組投入吉非替尼濃度為0、0.1μmol/l、1μmol/l、5μmol/l、10μmol/l并繼續(xù)孵育72小時后進行檢測。用劑量率為81.78cGy/min的60Co機進行放射治療,單純照射組分別以0、2Gy、4Gy、6Gy、8Gy的單次劑量照射后繼續(xù)在培養(yǎng)箱中孵育5到7天。藥物照射組,在放射治療30分鐘前先投入1μmol/l濃度的吉非替尼,再給予不同劑量照射培養(yǎng)5-7天后檢測以CCk-8測量細胞存活率；并用免疫熒光和免疫印跡分別檢測EGFR蛋白的表達水平。細胞集落實驗檢測吉非替尼對不同鼻咽癌細胞株放射敏感性的影響,設(shè)0Gy、2Gy、4Gy、6Gy、8Gy共5個劑量點。單純照射組和藥物照射組均射劑量大小以1000個細胞的密度接進行照射后,在37℃恒溫CO2培養(yǎng)箱中培養(yǎng)14天,在顯微鏡下觀測并計數(shù)細胞的克隆數(shù)(含有≥50個細胞的集落)。用單靶多擊數(shù)學(xué)模型擬合細胞存活曲線,分別計算各細胞株的放射增敏比(SER)。結(jié)果免疫熒光檢測和免疫印跡均顯示CNE1細胞和HONE1細胞內(nèi)均有較高EGFR表達,細胞存活實驗顯示,藥物照射組較單純照射組在低放射量組時既有顯著差異,(CNE1細胞株2Gy組P0.013,HONE1細胞株,2Gy組P0.008,4Gy組P0.02)。用單靶多擊數(shù)學(xué)模型擬合細胞存活曲線表示,CNE1細胞株的單純照射組細胞存活曲線的Do為2.069Gy,Dq為3.99Gy；藥物照射組Do為1.67Gy, Dq為2.49Gy,放射增敏比(SER)為1.23。HONE1細胞株照射組Do為2.0Gy,Dq為3.59Gy；藥物照射組Do為1.490Gy, Dq為2.71Gy,SER為1.35。結(jié)論EGFR在兩種細胞中均有較高表達,吉非替尼聯(lián)合放射照射較單純放射照射能提高鼻咽癌的治療效果。吉非替尼能增強鼻咽癌細胞株的放射敏感性。
[Abstract]:Objective the occurrence and development of nasopharyngeal carcinoma (NPC) are influenced and induced by a variety of etiological factors, including Epstein-Barr virus (EBV) infection, regional environmental factors, diet habits and family heredity. In addition, overexpression and variation of certain proteins and genes also play an important role in the development of nasopharyngeal carcinoma. Epidermal growth factor receptor (EGFR) is highly expressed in nasopharyngeal carcinoma (NPCs) cells, and is closely related to the occurrence and development of nasopharyngeal carcinoma (NPC). The purpose of this study was to study the effects of epidermal growth factor receptor (EGF) expression on the radiation of nasopharyngeal cells in vitro by comparing the killing effects of gifitinib alone, simple irradiation and combined irradiation with gefitinib on nasopharynx cells. To investigate the effect and possible mechanism of gefitinib on radiotherapy of nasopharyngeal carcinoma (NPC). Methods Human nasopharyngeal carcinoma (NPC) CNE1 and HONE1 cells in logarithmic phase were inoculated in 96-well plate and randomly divided into control group, gefitinib group, simple irradiation group and drug irradiation group. The concentration of gefitinib was 0. 1 渭 mol / L ~ (-1) 渭 mol / L ~ (-1) 渭 mol / L ~ (1) 5 渭 mol / L ~ (10) 渭 mol/l and incubated for 72 hours. Radiation therapy was performed with 60Co machine at dose rate of 81.78cGy/min. The irradiation group was incubated in incubator for 5 to 7 days after a single dose of 0 ~ 2 Gy ~ 4 Gy ~ 6 Gy ~ 8 Gy. In the drug irradiation group, 1 渭 mol/l concentration of gifitinib was injected 30 minutes before radiotherapy. After 5-7 days of different doses of irradiation, the survival rate of the cells was measured by CCk-8, and the expression of EGFR protein was detected by immunofluorescence and Western blotting. The effects of gefitinib on radiosensitivity of different nasopharyngeal carcinoma cell lines were detected by colony assay. After exposure to 1000 cells in the single irradiation group and drug irradiation group, the cells were cultured in a constant temperature CO2 incubator at 37 鈩，

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