本文選題：肝癌 + 細胞免疫治療��； 參考：《安徽理工大學(xué)》2017年碩士論文

【摘要】：目的:探討細胞免疫治療對肝癌患者外周血及局部病灶組織CXCR1、CXCR2、CXCL8表達的影響。方法:篩選典型的肝細胞肝癌患者35例,根據(jù)肝臟部位電子計算機斷層掃描(computerized tomographic scanning,CT)特征以及2010年美國癌癥聯(lián)合會公布的肝癌的TNM分期標(biāo)準(zhǔn)將其分為T2期24例,T3期11例;酶聯(lián)免疫吸附法分別測定患者血清中CXCL8、GP-73含量;以中性粒細胞分離液密度梯度分離純化外周血中性粒細胞,在PBNs中提取總RNA,后逆轉(zhuǎn)錄成cDNA并配制PCR體系進行擴增,凝膠電泳檢測肝癌患者體外周血及局部病灶組織CXCR1、CXCR2、CXCL8mRNA含量,以lgcDNA/lgGAPDH比值代表其最終mRNA水平。選取8例接受CIK細胞治療患者,進一步觀察T2、T3期患者CXCR1/2、CXCL8 mRNA含量變化。結(jié)果:肝癌患者血清內(nèi)CXCL8、GP-73含量分別是(315.66± 100.85)pg/mL、(214.84±49.11)ng/mL,與正常對照組相比均有顯著性差異(P0.0001);外周血PBNs內(nèi) CXCR1 mRNA、CXCR2 mRNA、CXCL8mRNA 分別為(1.1333 ±0.0551)lgcDNA/lgGAPDH、(0.8863 ±0.0605)lgcDNA/lgGAPDH、(1.2007±0.073)lgcDNA/lgGAPDH,與正常對照組相比均有顯著性差異(P0.0001);CIK細胞免疫治療后患者外周血PBNs內(nèi)CXCR1mRNA、CXCR2mRNA、CXCL8mRNA分別為(0.87±0.05631gcDNA/lgGAPDH、(0.76±0.0428)lgcDNA/lgGAPDH、(1.095±0.064)lgcDNA/lgGAPDH,與正常對照組相比均有顯著性差異(P0.0001);肝癌活檢組織蘇木精-伊紅染色法結(jié)果顯示:肝癌細胞體積明顯擴大,核形狀多變且不規(guī)則,核分裂增多,細胞互相重疊,分辨不清,瘤組織內(nèi)可見豐富的新生微血管;PCR檢測14例手術(shù)切除組織CXCR1/2、CXCL8 mRNA含量與正常對照相比,差異均具有統(tǒng)計學(xué)意義(z=6.528,P0.0001;t=9.583,P0.0001;t=7.832,P0.0001)。T2期肝癌患者的CXCR1/2、CXCL8 mRNA含量分別是(0.4462±0.0984)、(1.0509±0.1262)、(0.9032±0.1095)lgGAPDH/lgcDNA,T3 期肝癌患者的 CXCR1/2、CXCL8mRNA含量分別是(0.8296±0.1628)、(1.1361±0.1888)、(1.2649 ±0.2357)lgGAPDH/lgcDNA,統(tǒng)計學(xué)分析顯示 T2、T3 期肝癌患者CXCR1和CXCL8 mRNA表達差異有顯著性(t=4.341,P=0.0010;t=2.893,P=0.0135),T2、T3期肝癌患者CXCR2表達差異未見差異性(t=0.8215,P=0.4274)。部分切除肝癌組織HE染色見較多含淡染的紅細胞的管腔,提示病灶組織產(chǎn)生新生的血管。結(jié)論:(1)肝癌患者血清CXCL8、GP-73含量升高,且PBNs內(nèi)CXCL8和CXCR1/2 mRNA水平增加,提示血清中高表達的CXCL8與PBNs內(nèi)CXCL8 mRNA轉(zhuǎn)錄翻譯有關(guān)。(2)細胞免疫治療可下調(diào)PBNs內(nèi)CXCL8、CXCR1/2的過度表達,降低過度分泌CXCL8所致的炎癥反應(yīng)。(3)肝癌患者局部病灶組織CXCR1、CXCR2、CXCL8表達增加,提示PBNs參與局部病灶炎癥應(yīng)答,促進腫瘤細胞侵襲轉(zhuǎn)移。(4)抑制CXCR1、CXCR2過度表達,不僅可降低局部炎癥反應(yīng),還能抑制腫瘤細胞向周邊組織侵襲,似可作為抗肝癌細胞生長的新型分子靶位。
[Abstract]:Objective: to investigate the effect of cellular immunotherapy on the expression of CXCR1, CXCR2, CXCL8 in peripheral blood and local lesions of patients with hepatocellular carcinoma. Methods: Thirty-five patients with typical hepatocellular carcinoma were selected. According to the features of computerized tomographic scanning and the TNM staging criteria of HCC published by the American Cancer Federation in 2010, they were divided into 24 cases of T2 stage and 11 cases of T3 stage according to the features of computerized tomographic scanning. Serum CXCL8 GP-73 was determined by enzyme-linked immunosorbent assay (Elisa), peripheral blood neutrophils were isolated and purified by density gradient of neutrophils, total RNAs were extracted from PBNs, then reverse transcripted to cDNA and amplified by PCR system. The level of CXCR1, CXCR2, CXCL8 mRNA in peripheral blood and local lesions of patients with hepatocellular carcinoma was detected by gel electrophoresis, and the final mRNA level was represented by the ratio of lgcDNA/lgGAPDH. Eight patients receiving CIK cell therapy were selected to observe the changes of CXCR 1 / 2 C XCL8 mRNA in T 2 T 3 patients. 緇撴灉:鑲濈檶鎮(zhèn)ｈ，

本文編號：1866653