本文選題：喉腫瘤 + 西妥昔單抗�。� 參考：《山西醫(yī)科大學(xué)》2013年碩士論文

【摘要】：目的：西妥昔單抗（Cetuximab)是一種人鼠嵌合型IgG單克隆抗體，可與細(xì)胞表面的表皮生長因子受體(epidermal growth factor receptor, EGFR)特異性結(jié)合，競爭性地阻斷表皮生長因子和其配體的結(jié)合，從而阻斷腫瘤細(xì)胞增殖、轉(zhuǎn)移、侵襲以及血管生成等生物學(xué)效應(yīng)。Cetuximab與部分化療藥物或放射線對不同種類的腫瘤細(xì)胞具有協(xié)同殺傷的效應(yīng)，降低不同種類腫瘤細(xì)胞對Cetuximab的抵抗性，提高對Cetuximab的敏感性。本研究擬通過觀察西妥昔單抗聯(lián)合順鉑、放射線誘導(dǎo)人喉鱗癌細(xì)胞株H印-2的增殖、凋亡及細(xì)胞周期的影響，探討西妥昔單抗與順鉑、放射線聯(lián)合應(yīng)用對Hep-2細(xì)胞的殺傷效應(yīng)及對其調(diào)控機(jī)制的初步探討。 方法：1、體外培養(yǎng)人喉鱗狀細(xì)胞癌Hep-2細(xì)胞株，取對數(shù)生長期的細(xì)胞進(jìn)行試驗(yàn)。 2、采用CCK-8試劑盒（cell counting kit-8)分別檢測不同劑量的西妥昔單抗、順鉬、放射線對人喉鱗癌Hep-2細(xì)胞的12h、24h、48h、72h生長抑制率。 3、實(shí)驗(yàn)組分別給予西妥昔單抗1036ug/ml，順銷3ug/ml,放射線4Gy,順鉬3μg/ml+放射線4Gy,西妥昔單抗1036u g/ml’+順鉬3μg/ml，西妥昔單抗1036μg/ml+放射線4Gy，西妥昔單抗1036μg/ml+順柏3μg/m丨+放射線4Gy,同時(shí)設(shè)立陰性對照組（不添加任何藥物），分別作用于人喉鱗癌Hep-2細(xì)胞株24h，倒置顯微鏡觀察細(xì)胞形態(tài)，CCK-8試劑盒檢測細(xì)胞生長抑制率，流式細(xì)胞儀檢測不同干預(yù)方案對Hep-2凋亡率及細(xì)胞周期分布情況。 結(jié)果：1、不同劑量的西妥昔單抗、順鈾、放射線對Hep-2細(xì)胞均有抑制作用，并在一定濃度范圍內(nèi)呈時(shí)間-劑量依賴性，24h半數(shù)抑制濃度（halfmaximal inhibitory concentration, IC50)分別為兩妥昔單抗1036.84ii g/L、順鉬3.08μ g/ml、放射線4.18Gy; H印-2細(xì)胞對西妥昔單抗的生長抑制作用較敏感。 2、順鉑，放射線分別與西妥昔單抗聯(lián)合應(yīng)用時(shí)對Hep-2細(xì)胞的生長抑制率顯著高于順鉑、放射線單獨(dú)或聯(lián)合應(yīng)用（P0.001），具有協(xié)同殺傷效應(yīng)。 3、經(jīng)西妥昔單抗與放射線、西妥昔單抗與順鉑聯(lián)合作用24h后處于早期凋亡階段的Hep-2細(xì)胞分別為（21.92±3.00）%、（18.44±2.57）%，顯著高于陰性對照組與放射線、順鉑的單獨(dú)或聯(lián)合作用組（P0.001），提示西妥昔單抗對順鉑或放射線在體外誘導(dǎo)人喉癌Hep-2細(xì)胞株的凋亡具有顯著的協(xié)同作用。 4、單藥西妥昔單抗組中S期細(xì)胞所占比例較陰性對照組比例顯著增高（P0.001）；西妥昔單抗+放射線組（P=0.005），西妥昔單抗+順鉑+放射線組（P=0.002）中，G_0/G_1期細(xì)胞所占比例明顯高于單純放射線組。 結(jié)論：1、人喉鱗癌Hep-2細(xì)胞株對西妥昔單抗誘導(dǎo)的細(xì)胞凋亡敏感，在一定濃度范圍內(nèi)呈時(shí)間和劑量依賴性。 2、順鉑和/或放射線與西妥昔單抗聯(lián)用能顯著提高對Hep-2細(xì)胞的生長抑制作用，對Hep-2細(xì)胞的增殖具有協(xié)同抑制效應(yīng)。 3、西妥昔單抗與順鉑或放射線聯(lián)用能顯著提高Hep-2的凋亡率，提示西妥昔單抗對順鉑或放射線在體外誘導(dǎo)人喉癌Hep-2細(xì)胞株的凋亡具有顯著的協(xié)同作用。 4、西妥昔單抗可以通過將Hep-2細(xì)胞阻滯在細(xì)胞分裂的S期達(dá)到抑制細(xì)胞分裂阻止其增殖失控的作用。西妥昔單抗與放射線聯(lián)用亦可使人喉癌Hep-2細(xì)胞株阻滯在對放射線敏感的G_0/G_1期，體外可顯著誘導(dǎo)細(xì)胞凋亡，，從而對放射線具有較強(qiáng)的協(xié)同作用。這可能是順鉑、放射線與西妥昔單抗聯(lián)合應(yīng)用對Hep-2細(xì)胞發(fā)揮協(xié)同效應(yīng)的機(jī)制之一。
[Abstract]:Objective : To investigate the effect of cetuximab and cisplatin and radiation on the proliferation , apoptosis and cell cycle of human laryngeal squamous cell carcinoma cell line H - 2 , and to investigate the effect of cetuximab and cisplatin on the proliferation , apoptosis and cell cycle of human laryngeal squamous cell carcinoma cell line H - 2 .

Methods : 1 . Human laryngeal squamous cell carcinoma Hep2 cell line was cultured in vitro , and the cells with logarithmic growth were tested .

2 . CCK - 8 kit ( cell counting kit - 8 ) was used to detect the growth inhibition rate of cetuximab , cisplatin and radiation at 12 h , 24 h , 48 h and 72 h in human laryngeal squamous cell carcinoma .

3 . In the experimental group , 1036ug / ml , 3ug / ml , 4Gy , 1036u g / ml + radiation 4Gy , 1036u g / ml + irradiation 4Gy , 1036u g / ml + irradiation 4Gy , 1036u g / ml + cisplatin 3渭g / ml + radiation 4Gy were given .

Results : 1 . The inhibitory effect of different doses of cetuximab , UO2 and radiation on Hep2 cells was observed . The IC50 values were 106.84ii g / L , 3.08 渭g / ml , 4.18Gy and 4.18Gy , respectively , and the growth inhibition of cetuximab was more sensitive .

2 . The growth inhibition rate of cisplatin and radiation in combination with cetuximab was significantly higher than that of cisplatin , radiation alone or in combination ( P0.001 ) , which had synergistic killing effect .

3 . After the combined action of cetuximab and radiation , the combined action of cetuximab and cisplatin for 24 h was ( 21.92 鹵 3.00 ) % , ( 18.44 鹵 2.57 ) % , which was significantly higher than that of the negative control group ( P < 0.01 ) .

4 . The proportion of S - phase cells in the single - dose cetuximab group was significantly higher than that of the negative control group ( P0.001 ) .
In the cetuximab plus radiation group ( P = 0.005 ) , the percentage of G _ 0 / G _ 1 cells was significantly higher in the cetuximab plus cisplatin + radiation group ( P = 0.002 ) than in the simple radiation group .

Conclusion : 1 . Human laryngeal squamous cell carcinoma 2 2 cell line is sensitive to apoptosis induced by cetuximab , which is time and dose dependent within a certain concentration range .

2 , the combination of cisplatin and / or radiation and cetuximab can remarkably improve the growth inhibition effect on the Hep2 cells , and has a synergistic inhibitory effect on the proliferation of Hep2 cells .

3 . Combination of cetuximab and cisplatin or radiation could significantly increase the rate of apoptosis , suggesting that cetuximab had a significant synergistic effect on the apoptosis induced by cisplatin or radiation in vitro .

4 . The combination of cetuximab and radiation could arrest the cell division and prevent the cell division from being out of control .
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R739.65

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 萬文婷;李寧;劉靜;金林紅;;CCK-8法與MTT法檢測人前列腺癌PC3細(xì)胞活性的比較研究[J];時(shí)珍國醫(yī)國藥;2010年12期

2 金晶,高黎,徐國鎮(zhèn),袁智勇,王穎杰,李素艷;單純放療或單純手術(shù)治療早期聲門型喉癌[J];中華放射腫瘤學(xué)雜志;2005年01期

本文編號：2064656