γδT细胞抑制胰腺癌细胞增长的作用及其机制研究

发布时间：2018-11-05 07:41

【摘要】：【目的】在前期体外实验研究的基础上。继续深入探讨NKG2D受体和其相关配体ULBP在γδT细胞抑制胰腺癌裸鼠移植瘤生长中的作用。检测胰腺癌组织中的ULBP表达。人体外周血中ULBP的含量对胰腺癌诊断价值。【方法】利用抗体吸附法从健康志愿者外周血中分离培养出γδT细胞。将人胰腺癌细胞株PANC-1注入BALB/C裸鼠背部皮下以建立裸鼠胰腺癌动物模型。把裸鼠随机地分成吉西他滨治疗组,对照组和γδT细胞组。体外分离培养γδT细胞,扩增后经裸鼠尾静脉注入。每周进行2次裸鼠重量和肿瘤体积的测量。随后手术摘除肿瘤组织并计算相对肿瘤体积和肿瘤抑制率。运用WB和qPCR检测ULBPs在三组裸鼠胰腺癌移植瘤中的表达。IHC检测30个胰腺癌患者肿瘤组织的ULBPs的表达情况。ELISA法检测胰腺癌患者及健康志愿者的外周血中ULBPs和CA199的表达水平及外周血中ULBPs对胰腺癌诊断价值。【结果】胰腺癌肿瘤生长在γδT细胞治疗组受到明显抑制(P=0.003)。并且其抑瘤率和吉西他滨治疗组的肿瘤抑瘤率无明显差别。人胰腺癌组织的IHC实验及裸鼠瘤组织的WB实验都显示ULBP在胰腺癌组织中过表达。此外,WB实验表明经过γδT的治疗后,ULBP-1和ULBP-3的表达水平会下降(P=0.025,P=0.019)。可溶性ULBP2在胰腺癌患者外周血中的表达要比健康人群外周血中的含量高;可溶性ULBP2有助于CA19-9对胰腺癌的诊断[AUC=0.88(95%CI:0.807~0.943);AUC=0.84(95%CI:0.761~0.917)]。【结论】γδT细胞对胰腺癌生长增值具有显著的抑制作用,其发挥作用的机制可能与ULBP异常表达相关。可溶性ULBP2联合CA199诊断胰腺癌的效果优于单独CA199。
[Abstract]:[objective] on the basis of in vitro experimental study. To further investigate the role of NKG2D receptor and its associated ligand ULBP in inhibiting the growth of pancreatic carcinoma xenografts in nude mice by 纬 未 T cells. The expression of ULBP in pancreatic carcinoma was detected. [methods] 纬 未 T cells were isolated from the peripheral blood of healthy volunteers by antibody adsorption method. Human pancreatic cancer cell line PANC-1 was injected subcutaneously into the back of BALB/C nude mice to establish nude mice pancreatic cancer model. Nude mice were randomly divided into gemcitabine group, control group and 纬 未 T cell group. 纬 未 T cells were isolated and cultured in vitro. The weight and tumor volume of nude mice were measured twice a week. Then the tumor tissue was removed and the relative tumor volume and tumor inhibition rate were calculated. WB and qPCR were used to detect the expression of ULBPs in transplanted pancreatic carcinoma in three groups of nude mice. IHC was used to detect the expression of ULBPs in tumor tissues of 30 patients with pancreatic cancer. ELISA method was used to detect the expression of ULBPs and CA199 in peripheral blood of pancreatic cancer patients and healthy volunteers. [results] Pancreatic carcinoma tumor growth was significantly inhibited in 纬 未 T cell therapy group (P0. 003). There was no significant difference in tumor inhibition rate between gemcitabine group and gemcitabine group. IHC assay in human pancreatic carcinoma and WB assay in nude mice showed that ULBP was overexpressed in pancreatic carcinoma. In addition, WB assay showed that after 纬 未 T treatment, the expression of ULBP-1 and ULBP-3 decreased (P0. 025, P0. 019). The expression of soluble ULBP2 in peripheral blood of pancreatic cancer patients was higher than that of healthy controls, and soluble ULBP2 was helpful to the diagnosis of pancreatic cancer by CA19-9 [AUC=0.88 (95%CI:0.807~0.943)]. AUC=0.84 (95%CI:0.761~0.917)]. [conclusion] 纬 未 T cells have a significant inhibitory effect on the growth and proliferation of pancreatic carcinoma, and the mechanism of its action may be related to the abnormal expression of ULBP. The effect of soluble ULBP2 combined with CA199 in the diagnosis of pancreatic cancer is better than that of CA199. alone.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.9
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