自然杀伤T细胞数量分布变化与肝癌转移的相关性研究

发布时间：2018-05-20 08:35

  本文选题：肝癌 + NKT淋巴　； 参考：《青岛大学》2017年硕士论文

【摘要】：目的:通过研究肝癌组织中自然杀伤T细胞(Natural killer T cells,NKT)的数量分布变化与肝癌转移的相关性,进一步明确NKT细胞与肝癌疾病进展期的变化规律,为肝癌的预后判断与治疗提供科学依据,增加NKT细胞在肝脏疾病中的临床数据,为进一步探索肝脏微环境与NKT细胞之间的相互影响,提供实验依据并探索肝癌免疫治疗新方法。研究对象及方法:1、研究对象:收集青岛大学附属医院肝胆外科、普外科2016.01~2016.05年间手术切除并经病理证实的肝癌标本共64例(术前未行放、化疗),标本根据全国肝癌协会会议拟定的分期为标准进行临床分期,均经4g/L的中性甲醛固定,常规石蜡包埋,4um厚常规切片,苏木精-伊红(HE)染色和复核病理诊断。另取非肝癌手术的正常肝组织15例作为对照。2、方法:选择小鼠抗人CD3单抗和小鼠抗人CD56单抗,采用双重免疫组织化学SP法对切片进行抗原染色,DAB和AEC显色剂分别显色后,光学显微镜下分别计数在正常肝脏组织,临床不同分期肝癌组织中NKT淋巴细胞的数量及分布情况,运用统计学方法分析NKT淋巴细胞的数量分布变化与肝癌细胞迁移的相关性。切片中能同时表达CD3、CD56抗原分子的淋巴细胞为阳性NKT细胞。结果:1、15例正常肝脏组织NKT细胞单个高倍视野平均数为18.3571 8.3265,且NKT细胞多集中在门管区微血管和肝血窦内。2、28例Ⅰa-Ⅱa期肝癌组织NKT细胞单个高倍视野平均数为9.2438 2.5894,随病程进展肝脏内微环境的变化NKT细胞数量逐渐下降。3、36例Ⅱb-Ⅲb肝癌组织NKT细胞单个高倍视野平均数为7.6587 5.9854。晚期肝癌NKT细胞数量进一步下降,视野内NKT数量很少。4、肝癌组织中(未分期)自然杀伤性T淋巴细胞的数量明显低于正常肝组织内NKT细胞的数量,具有明显差异P0.05(P=0.037)5、不同分期肝癌中Ⅰa期-Ⅱa期与Ⅱb期-Ⅲb期的NKT细胞数量有明显差异P0.05(P=0.025),且癌细胞侵袭肝脏的程度越高,NKT细胞数量越少。结论:(1)利用不同的免疫细胞化学显色剂,分别显色NKT细胞膜上表达的CD3和CD56抗原,对NKT细胞进行识别和定位。(2)正常肝脏组织NKT细胞细胞多集中在门管区微血管内和肝血窦内。(3)肝癌组织中(未分期)自然杀伤性T淋巴细胞的数量明显低于正常肝组织内NKT细胞的数量。(4)NKT细胞数量随肝癌病程进展逐渐下降。癌细胞侵袭肝脏的程度越高,NKT细胞数量越少。
[Abstract]:Objective: to study the relationship between the quantity distribution of natural killer T cells and the metastasis of hepatocellular carcinoma (HCC), and to study the relationship between NKT cells and the progression of HCC. It can provide scientific basis for prognosis and treatment of liver cancer, increase the clinical data of NKT cells in liver diseases, further explore the interaction between liver microenvironment and NKT cells, provide experimental basis and explore a new method of immunotherapy for liver cancer. Subjects and methods: study subjects: we collected 64 specimens of hepatobiliary surgery from the affiliated Hospital of Qingdao University, which were surgically resected and pathologically proved by general surgery during the period of 2016.01and 2016.05. All the specimens were fixed by neutral formaldehyde of 4g/L, paraffin-embedded 4um thick conventional sections, hematoxylin-eosin (HEH) staining and pathological diagnosis. Another 15 cases of normal liver tissue were taken as control group. Methods: mouse anti-human CD3 monoclonal antibody and mouse anti-human CD56 monoclonal antibody were selected, and the sections were stained by antigen staining and AEC respectively by double immunohistochemistry SP method. The number and distribution of NKT lymphocytes in normal liver tissues and clinical HCC tissues were counted under optical microscope. The correlation between the number distribution of NKT lymphocytes and the migration of HCC cells was analyzed by statistical method. The lymphocytes which could express CD3 + CD56 antigen were positive NKT cells. Results the average of single high-power field of NKT cells in 15 normal liver tissues was 18.3571 8.3265, and the average of NKT cells was 9.2438 2.5894 in 28 patients with stage 鈪，

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