细胞免疫治疗对肝癌患者CXCR1、CXCR2、CXCL8表达的影响

发布时间：2018-05-09 16:03

  本文选题：肝癌 + 细胞免疫治疗　； 参考：《安徽理工大学》2017年硕士论文

【摘要】：目的:探讨细胞免疫治疗对肝癌患者外周血及局部病灶组织CXCR1、CXCR2、CXCL8表达的影响。方法:筛选典型的肝细胞肝癌患者35例,根据肝脏部位电子计算机断层扫描(computerized tomographic scanning,CT)特征以及2010年美国癌症联合会公布的肝癌的TNM分期标准将其分为T2期24例,T3期11例;酶联免疫吸附法分别测定患者血清中CXCL8、GP-73含量;以中性粒细胞分离液密度梯度分离纯化外周血中性粒细胞,在PBNs中提取总RNA,后逆转录成cDNA并配制PCR体系进行扩增,凝胶电泳检测肝癌患者体外周血及局部病灶组织CXCR1、CXCR2、CXCL8mRNA含量,以lgcDNA/lgGAPDH比值代表其最终mRNA水平。选取8例接受CIK细胞治疗患者,进一步观察T2、T3期患者CXCR1/2、CXCL8 mRNA含量变化。结果:肝癌患者血清内CXCL8、GP-73含量分别是(315.66± 100.85)pg/mL、(214.84±49.11)ng/mL,与正常对照组相比均有显著性差异(P0.0001);外周血PBNs内 CXCR1 mRNA、CXCR2 mRNA、CXCL8mRNA 分别为(1.1333 ±0.0551)lgcDNA/lgGAPDH、(0.8863 ±0.0605)lgcDNA/lgGAPDH、(1.2007±0.073)lgcDNA/lgGAPDH,与正常对照组相比均有显著性差异(P0.0001);CIK细胞免疫治疗后患者外周血PBNs内CXCR1mRNA、CXCR2mRNA、CXCL8mRNA分别为(0.87±0.05631gcDNA/lgGAPDH、(0.76±0.0428)lgcDNA/lgGAPDH、(1.095±0.064)lgcDNA/lgGAPDH,与正常对照组相比均有显著性差异(P0.0001);肝癌活检组织苏木精-伊红染色法结果显示:肝癌细胞体积明显扩大,核形状多变且不规则,核分裂增多,细胞互相重叠,分辨不清,瘤组织内可见丰富的新生微血管;PCR检测14例手术切除组织CXCR1/2、CXCL8 mRNA含量与正常对照相比,差异均具有统计学意义(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含量分别是(0.4462±0.0984)、(1.0509±0.1262)、(0.9032±0.1095)lgGAPDH/lgcDNA,T3 期肝癌患者的 CXCR1/2、CXCL8mRNA含量分别是(0.8296±0.1628)、(1.1361±0.1888)、(1.2649 ±0.2357)lgGAPDH/lgcDNA,统计学分析显示 T2、T3 期肝癌患者CXCR1和CXCL8 mRNA表达差异有显著性(t=4.341,P=0.0010;t=2.893,P=0.0135),T2、T3期肝癌患者CXCR2表达差异未见差异性(t=0.8215,P=0.4274)。部分切除肝癌组织HE染色见较多含淡染的红细胞的管腔,提示病灶组织产生新生的血管。结论:(1)肝癌患者血清CXCL8、GP-73含量升高,且PBNs内CXCL8和CXCR1/2 mRNA水平增加,提示血清中高表达的CXCL8与PBNs内CXCL8 mRNA转录翻译有关。(2)细胞免疫治疗可下调PBNs内CXCL8、CXCR1/2的过度表达,降低过度分泌CXCL8所致的炎症反应。(3)肝癌患者局部病灶组织CXCR1、CXCR2、CXCL8表达增加,提示PBNs参与局部病灶炎症应答,促进肿瘤细胞侵袭转移。(4)抑制CXCR1、CXCR2过度表达,不仅可降低局部炎症反应,还能抑制肿瘤细胞向周边组织侵袭,似可作为抗肝癌细胞生长的新型分子靶位。
[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. 缁撴灉:鑲濈檶鎮ｈ，

本文编号：1866653