【摘要】：目的:結(jié)腸癌作為常見的消化道惡性腫瘤,在我國的發(fā)病率和死亡率均呈逐年上升趨勢。結(jié)腸癌由于術(shù)后易復(fù)發(fā),放化療耐受等特點(diǎn),目前臨床上的治療效果都不理想。因此,需要探索出新的結(jié)腸癌治療策略。腫瘤干細(xì)胞(cancer stem cells,CSCs)是腫瘤組織中存在的較少的具有自我更新能力并能無限增殖、多向分化并對(duì)放化療不敏感的細(xì)胞亞群,與腫瘤的發(fā)生、發(fā)展和各種生物學(xué)抗性密切相關(guān)。腫瘤干細(xì)胞理論的提出使人們認(rèn)識(shí)到,以腫瘤干細(xì)胞為靶點(diǎn)的抗腫瘤治療策略為徹底治愈腫瘤帶來了可能。樹突狀細(xì)胞(dendritic cell,DC)是目前已知的生物體內(nèi)功能最強(qiáng)的專職抗原提呈細(xì)胞(antigen presenting cell,APC),能夠特異性地誘導(dǎo)機(jī)體免疫應(yīng)答的發(fā)生。PD-L1作為負(fù)性免疫共刺激分子在腫瘤免疫逃逸過程中起到重要作用,抗PD-L1單抗能與腫瘤微環(huán)境中表達(dá)的PD-L1相結(jié)合,減弱PD-L1介導(dǎo)的免疫逃逸作用,從而增強(qiáng)DC疫苗介導(dǎo)的的免疫治療效果。因此,本實(shí)驗(yàn)以CD44作為分選標(biāo)志物,從小鼠結(jié)腸癌細(xì)胞系CT-26分選出CD44+細(xì)胞作為小鼠腫瘤干細(xì)胞,擬將CD44+CT-26細(xì)胞完全抗原致敏的DC疫苗與抗PD-L1單抗聯(lián)用,探討這種聯(lián)合治療策略能否有效地治療小鼠結(jié)腸癌,為結(jié)腸癌靶向治療提供實(shí)驗(yàn)依據(jù)。方法:免疫磁珠法分選獲得CD44+CT-26細(xì)胞,用含20 ng/ml EGF、20 ng/ml b FGF、2%B27的DMEM/F12無血清腫瘤干細(xì)胞培養(yǎng)基培養(yǎng);從小鼠骨髓中獲得DC,將CT-26細(xì)胞裂解物和CD44+CT-26細(xì)胞裂解物分別作為抗原負(fù)載DC,獲得DCCT-26和DCSC疫苗;然后,分別與抗PD-L1單抗聯(lián)用,進(jìn)行結(jié)腸癌荷瘤小鼠免疫治療實(shí)驗(yàn)。分析小鼠的生存狀態(tài),腫瘤生長情況,小鼠脾來源T細(xì)胞對(duì)結(jié)腸癌細(xì)胞的殺傷作用。結(jié)果:(1)與Control組和αPD-L1組相比,DCSC+αPD-L1組小鼠的腫瘤生長顯著受到抑制(P0.05);腫瘤重量顯著減小(P0.05);小鼠脾來源CTL對(duì)CT-26細(xì)胞的殺傷顯著增強(qiáng)(P0.05)。(2)與DCSC組相比,除小鼠脾來源CTL對(duì)CT-26細(xì)胞的殺傷顯著增強(qiáng)(P0.05)外,DCSC+αPD-L1組與DCSC組其他指標(biāo)無顯著性差異。(3)與DCCT-26組相比,除小鼠腫瘤重量顯著減小(P0.05)外,DCSC組與DCCT-26組其他指標(biāo)無顯著性差異。(4)與DCCT-26+αPD-L1組相比,DCSC+αPD-L1組與DCCT-26+αPD-L1組其他指標(biāo)無顯著性差異。結(jié)論:在BALB/c小鼠結(jié)腸癌模型中,CD44+CT26細(xì)胞完全抗原負(fù)載的DC疫苗與抗PD-L1單抗聯(lián)用可以產(chǎn)生有效地抗腫瘤免疫治療效果,抑制腫瘤生長,增強(qiáng)對(duì)腫瘤細(xì)胞的殺傷效果。但是與腫瘤細(xì)胞抗原致敏的DC疫苗相比,抗腫瘤免疫治療優(yōu)勢不明顯。
[Abstract]:%E7%9B%AE%E7%9A%84%3A%E7%BB%93%E8%82%A0%E7%99%8C%E4%BD%9C%E4%B8%BA%E5%B8%B8%E8%A7%81%E7%9A%84%E6%B6%88%E5%8C%96%E9%81%93%E6%81%B6%E6%80%A7%E8%82%BF%E7%98%A4%2C%E5%9C%A8%E6%88%91%E5%9B%BD%E7%9A%84%E5%8F%91%E7%97%85%E7%8E%87%E5%92%8C%E6%AD%BB%E4%BA%A1%E7%8E%87%E5%9D%87%E5%91%88%E9%80%90%E5%B9%B4%E4%B8%8A%E5%8D%87%E8%B6%8B%E5%8A%BF.%E7%BB%93%E8%82%A0%E7%99%8C%E7%94%B1%E4%BA%8E%E6%9C%AF%E5%90%8E%E6%98%93%E5%A4%8D%E5%8F%91%2C%E6%94%BE%E5%8C%96%E7%96%97%E8%80%90%E5%8F%97%E7%AD%89%E7%89%B9%E7%82%B9%2C%E7%9B%AE%E5%89%8D%E4%B8%B4%E5%BA%8A%E4%B8%8A%E7%9A%84%E6%B2%BB%E7%96%97%E6%95%88%E6%9E%9C%E9%83%BD%E4%B8%8D%E7%90%86%E6%83%B3.%E5%9B%A0%E6%AD%A4%2C%E9%9C%80%E8%A6%81%E6%8E%A2%E7%B4%A2%E5%87%BA%E6%96%B0%E7%9A%84%E7%BB%93%E8%82%A0%E7%99%8C%E6%B2%BB%E7%96%97%E7%AD%96%E7%95%A5.%E8%82%BF%E7%98%A4%E5%B9%B2%E7%BB%86%E8%83%9E%28cancer%20stem%20cells%2CCSCs%29%E6%98%AF%E8%82%BF%E7%98%A4%E7%BB%84%E7%BB%87%E4%B8%AD%E5%AD%98%E5%9C%A8%E7%9A%84%E8%BE%83%E5%B0%91%E7%9A%84%E5%85%B7%E6%9C%89%E8%87%AA%E6%88%91%E6%9B%B4%E6%96%B0%E8%83%BD%E5%8A%9B%E5%B9%B6%E8%83%BD%E6%97%A0%E9%99%90%E5%A2%9E%E6%AE%96%2C%E5%A4%9A%E5%90%91%E5%88%86%E5%8C%96%E5%B9%B6%E5%AF%B9%E6%94%BE%E5%8C%96%E7%96%97%E4%B8%8D%E6%95%8F%E6%84%9F%E7%9A%84%E7%BB%86%E8%83%9E%E4%BA%9A%E7%BE%A4%2C%E4%B8%8E%E8%82%BF%E7%98%A4%E7%9A%84%E5%8F%91%E7%94%9F%2C%E5%8F%91%E5%B1%95%E5%92%8C%E5%90%84%E7%A7%8D%E7%94%9F%E7%89%A9%E5%AD%A6%E6%8A%97%E6%80%A7%E5%AF%86%E5%88%87%E7%9B%B8%E5%85%B3.%E8%82%BF%E7%98%A4%E5%B9%B2%E7%BB%86%E8%83%9E%E7%90%86%E8%AE%BA%E7%9A%84%E6%8F%90%E5%87%BA%E4%BD%BF%E4%BA%BA%E4%BB%AC%E8%AE%A4%E8%AF%86%E5%88%B0%2C%E4%BB%A5%E8%82%BF%E7%98%A4%E5%B9%B2%E7%BB%86%E8%83%9E%E4%B8%BA%E9%9D%B6%E7%82%B9%E7%9A%84%E6%8A%97%E8%82%BF%E7%98%A4%E6%B2%BB%E7%96%97%E7%AD%96%E7%95%A5%E4%B8%BA%E5%BD%BB%E5%BA%95%E6%B2%BB%E6%84%88%E8%82%BF%E7%98%A4%E5%B8%A6%E6%9D%A5%E4%BA%86%E5%8F%AF%E8%83%BD.%E6%A0%91%E7%AA%81%E7%8A%B6%E7%BB%86%E8%83%9E%28dendritic%20cell%2CDC%29%E6%98%AF%E7%9B%AE%E5%89%8D%E5%B7%B2%E7%9F%A5%E7%9A%84%E7%94%9F%E7%89%A9%E4%BD%93%E5%86%85%E5%8A%9F%E8%83%BD%E6%9C%80%E5%BC%BA%E7%9A%84%E4%B8%93%E8%81%8C%E6%8A%97%E5%8E%9F%E6%8F%90%E5%91%88%E7%BB%86%E8%83%9E%28antigen%20presenting%20cell%2CAPC%29%2C%E8%83%BD%E5%A4%9F%E7%89%B9%E5%BC%82%E6%80%A7%E5%9C%B0%E8%AF%B1%E5%AF%BC%E6%9C%BA%E4%BD%93%E5%85%8D%E7%96%AB%E5%BA%94%E7%AD%94%E7%9A%84%E5%8F%91%E7%94%9F.PD-L1%E4%BD%9C%E4%B8%BA%E8%B4%9F%E6%80%A7%E5%85%8D%E7%96%AB%E5%85%B1%E5%88%BA%E6%BF%80%E5%88%86%E5%AD%90%E5%9C%A8%E8%82%BF%E7%98%A4%E5%85%8D%E7%96%AB%E9%80%83%E9%80%B8%E8%BF%87%E7%A8%8B%E4%B8%AD%E8%B5%B7%E5%88%B0%E9%87%8D%E8%A6%81%E4%BD%9C%E7%94%A8%2C%E6%8A%97PD-L1%E5%8D%95%E6%8A%97%E8%83%BD%E4%B8%8E%E8%82%BF%E7%98%A4%E5%BE%AE%E7%8E%AF%E5%A2%83%E4%B8%AD%E8%A1%A8%E8%BE%BE%E7%9A%84PD-L1%E7%9B%B8%E7%BB%93%E5%90%88%2C%E5%87%8F%E5%BC%B1PD-L1%E4%BB%8B%E5%AF%BC%E7%9A%84%E5%85%8D%E7%96%AB%E9%80%83%E9%80%B8%E4%BD%9C%E7%94%A8%2C%E4%BB%8E%E8%80%8C%E5%A2%9E%E5%BC%BADC%E7%96%AB%E8%8B%97%E4%BB%8B%E5%AF%BC%E7%9A%84%E7%9A%84%E5%85%8D%E7%96%AB%E6%B2%BB%E7%96%97%E6%95%88%E6%9E%9C.%E5%9B%A0%E6%AD%A4%2C%E6%9C%AC%E5%AE%9E%E9%AA%8C%E4%BB%A5CD44%E4%BD%9C%E4%B8%BA%E5%88%86%E9%80%89%E6%A0%87%E5%BF%97%E7%89%A9%2C%E4%BB%8E%E5%B0%8F%E9%BC%A0%E7%BB%93%E8%82%A0%E7%99%8C%E7%BB%86%E8%83%9E%E7%B3%BBCT-26%E5%88%86%E9%80%89%E5%87%BACD44%20%E7%BB%86%E8%83%9E%E4%BD%9C%E4%B8%BA%E5%B0%8F%E9%BC%A0%E8%82%BF%E7%98%A4%E5%B9%B2%E7%BB%86%E8%83%9E%2C%E6%8B%9F%E5%B0%86CD44%20CT-26%E7%BB%86%E8%83%9E%E5%AE%8C%E5%85%A8%E6%8A%97%E5%8E%9F%E8%87%B4%E6%95%8F%E7%9A%84DC%E7%96%AB%E8%8B%97%E4%B8%8E%E6%8A%97PD-L1%E5%8D%95%E6%8A%97%E8%81%94%E7%94%A8%2C%E6%8E%A2%E8%AE%A8%E8%BF%99%E7%A7%8D%E8%81%94%E5%90%88%E6%B2%BB%E7%96%97%E7%AD%96%E7%95%A5%E8%83%BD%E5%90%A6%E6%9C%89%E6%95%88%E5%9C%B0%E6%B2%BB%E7%96%97%E5%B0%8F%E9%BC%A0%E7%BB%93%E8%82%A0%E7%99%8C%2C%E4%B8%BA%E7%BB%93%E8%82%A0%E7%99%8C%E9%9D%B6%E5%90%91%E6%B2%BB%E7%96%97%E6%8F%90%E4%BE%9B%E5%AE%9E%E9%AA%8C%E4%BE%9D%E6%8D%AE.%E6%96%B9%E6%B3%95%3A%E5%85%8D%E7%96%AB%E7%A3%81%E7%8F%A0%E6%B3%95%E5%88%86%E9%80%89%E8%8E%B7%E5%BE%97CD44%20CT-26%E7%BB%86%E8%83%9E%2C%E7%94%A8%E5%90%AB20%20ng%2Fml%20EGF%2C20%20ng%2Fml%20b%20FGF%2C2%B27%E7%9A%84DMEM%2FF12%E6%97%A0%E8%A1%80%E6%B8%85%E8%82%BF%E7%98%A4%E5%B9%B2%E7%BB%86%E8%83%9E%E5%9F%B9%E5%85%BB%E5%9F%BA%E5%9F%B9%E5%85%BB%3B%E4%BB%8E%E5%B0%8F%E9%BC%A0%E9%AA%A8%E9%AB%93%E4%B8%AD%E8%8E%B7%E5%BE%97DC%2C%E5%B0%86CT-26%E7%BB%86%E8%83%9E%E8%A3%82%E8%A7%A3%E7%89%A9%E5%92%8CCD44%20CT-26%E7%BB%86%E8%83%9E%E8%A3%82%E8%A7%A3%E7%89%A9%E5%88%86%E5%88%AB%E4%BD%9C%E4%B8%BA%E6%8A%97%E5%8E%9F%E8%B4%9F%E8%BD%BDDC%2C%E8%8E%B7%E5%BE%97DCCT-26%E5%92%8CDCSC%E7%96%AB%E8%8B%97%3B%E7%84%B6%E5%90%8E%2C%E5%88%86%E5%88%AB%E4%B8%8E%E6%8A%97PD-L1%E5%8D%95%E6%8A%97%E8%81%94%E7%94%A8%2C%E8%BF%9B%E8%A1%8C%E7%BB%93%E8%82%A0%E7%99%8C%E8%8D%B7%E7%98%A4%E5%B0%8F%E9%BC%A0%E5%85%8D%E7%96%AB%E6%B2%BB%E7%96%97%E5%AE%9E%E9%AA%8C.%E5%88%86%E6%9E%90%E5%B0%8F%E9%BC%A0%E7%9A%84%E7%94%9F%E5%AD%98%E7%8A%B6%E6%80%81%2C%E8%82%BF%E7%98%A4%E7%94%9F%E9%95%BF%E6%83%85%E5%86%B5%2C%E5%B0%8F%E9%BC%A0%E8%84%BE%E6%9D%A5%E6%BA%90T%E7%BB%86%E8%83%9E%E5%AF%B9%E7%BB%93%E8%82%A0%E7%99%8C%E7%BB%86%E8%83%9E%E7%9A%84%E6%9D%80%E4%BC%A4%E4%BD%9C%E7%94%A8.%E7%BB%93%E6%9E%9C%3A%281%29%E4%B8%8EControl%E7%BB%84%E5%92%8C%CE%B1PD-L1%E7%BB%84%E7%9B%B8%E6%AF%94%2CDCSC%20%CE%B1PD-L1%E7%BB%84%E5%B0%8F%E9%BC%A0%E7%9A%84%E8%82%BF%E7%98%A4%E7%94%9F%E9%95%BF%E6%98%BE%E8%91%97%E5%8F%97%E5%88%B0%E6%8A%91%E5%88%B6%28P0.05%29%3B%E8%82%BF%E7%98%A4%E9%87%8D%E9%87%8F%E6%98%BE%E8%91%97%E5%87%8F%E5%B0%8F%28P0.05%29%3B%E5%B0%8F%E9%BC%A0%E8%84%BE%E6%9D%A5%E6%BA%90CTL%E5%AF%B9CT-26%E7%BB%86%E8%83%9E%E7%9A%84%E6%9D%80%E4%BC%A4%E6%98%BE%E8%91%97%E5%A2%9E%E5%BC%BA%28P0.05%29.%282%29%E4%B8%8EDCSC%E7%BB%84%E7%9B%B8%E6%AF%94%2C%E9%99%A4%E5%B0%8F%E9%BC%A0%E8%84%BE%E6%9D%A5%E6%BA%90CTL%E5%AF%B9CT-26%E7%BB%86%E8%83%9E%E7%9A%84%E6%9D%80%E4%BC%A4%E6%98%BE%E8%91%97%E5%A2%9E%E5%BC%BA%28P0.05%29%E5%A4%96%2CDCSC%20%CE%B1PD-L1%E7%BB%84%E4%B8%8EDCSC%E7%BB%84%E5%85%B6%E4%BB%96%E6%8C%87%E6%A0%87%E6%97%A0%E6%98%BE%E8%91%97%E6%80%A7%E5%B7%AE%E5%BC%82.%283%29%E4%B8%8EDCCT-26%E7%BB%84%E7%9B%B8%E6%AF%94%2C%E9%99%A4%E5%B0%8F%E9%BC%A0%E8%82%BF%E7%98%A4%E9%87%8D%E9%87%8F%E6%98%BE%E8%91%97%E5%87%8F%E5%B0%8F%28P0.05%29%E5%A4%96%2CDCSC%E7%BB%84%E4%B8%8EDCCT-26%E7%BB%84%E5%85%B6%E4%BB%96%E6%8C%87%E6%A0%87%E6%97%A0%E6%98%BE%E8%91%97%E6%80%A7%E5%B7%AE%E5%BC%82.%284%29%E4%B8%8EDCCT-26%20%CE%B1PD-L1%E7%BB%84%E7%9B%B8%E6%AF%94%2CDCSC%20%CE%B1PD-L1%E7%BB%84%E4%B8%8EDCCT-26%20%CE%B1PD-L1%E7%BB%84%E5%85%B6%E4%BB%96%E6%8C%87%E6%A0%87%E6%97%A0%E6%98%BE%E8%91%97%E6%80%A7%E5%B7%AE%E5%BC%82.%E7%BB%93%E8%AE%BA%3A%E5%9C%A8BALB%2Fc%E5%B0%8F%E9%BC%A0%E7%BB%93%E8%82%A0%E7%99%8C%E6%A8%A1%E5%9E%8B%E4%B8%AD%2CCD44%20CT26%E7%BB%86%E8%83%9E%E5%AE%8C%E5%85%A8%E6%8A%97%E5%8E%9F%E8%B4%9F%E8%BD%BD%E7%9A%84DC%E7%96%AB%E8%8B%97%E4%B8%8E%E6%8A%97PD-L1%E5%8D%95%E6%8A%97%E8%81%94%E7%94%A8%E5%8F%AF%E4%BB%A5%E4%BA%A7%E7%94%9F%E6%9C%89%E6%95%88%E5%9C%B0%E6%8A%97%E8%82%BF%E7%98%A4%E5%85%8D%E7%96%AB%E6%B2%BB%E7%96%97%E6%95%88%E6%9E%9C%2C%E6%8A%91%E5%88%B6%E8%82%BF%E7%98%A4%E7%94%9F%E9%95%BF%2C%E5%A2%9E%E5%BC%BA%E5%AF%B9%E8%82%BF%E7%98%A4%E7%BB%86%E8%83%9E%E7%9A%84%E6%9D%80%E4%BC%A4%E6%95%88%E6%9E%9C.%E4%BD%86%E6%98%AF%E4%B8%8E%E8%82%BF%E7%98%A4%E7%BB%86%E8%83%9E%E6%8A%97%E5%8E%9F%E8%87%B4%E6%95%8F%E7%9A%84DC%E7%96%AB%E8%8B%97%E7%9B%B8%E6%AF%94%2C%E6%8A%97%E8%82%BF%E7%98%A4%E5%85%8D%E7%96%AB%E6%B2%BB%E7%96%97%E4%BC%98%E5%8A%BF%E4%B8%8D%E6%98%8E%E6%98%BE.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R735.35

【參考文獻(xiàn)】

相關(guān)期刊論文 前5條

1 Wanqing Chen;Rongshou Zheng;Tingting Zuo;Hongmei Zeng;Siwei Zhang;Jie He;;National cancer incidence and mortality in China, 2012[J];Chinese Journal of Cancer Research;2016年01期

2 劉見榮;侯風(fēng)剛;;結(jié)腸癌治療概況[J];遼寧中醫(yī)藥大學(xué)學(xué)報(bào);2014年02期

3 陳瓊;劉志才;程蘭平;宋國慧;孫喜斌;鄭榮壽;張思維;陳萬青;;2003～2007年中國結(jié)直腸癌發(fā)病與死亡分析[J];中國腫瘤;2012年03期

4 Mabood Qureshi;Jim Xiang;;Dendritic Cell-Derived Exosomes Stimulate Stronger CD8~+ CTL Responses and Antitumor Immunity than TVimor Cell-Derived Exosomes[J];Cellular & Molecular Immunology;2006年03期

5 王正康;黃莛庭;;結(jié)腸癌根治術(shù)的合理切除范圍(文獻(xiàn)綜述)[J];國外醫(yī)學(xué).外科學(xué)分冊;1989年01期

，

本文編號(hào)：2265614