【摘要】：【目的】在前期體外實驗研究的基礎上。繼續(xù)深入探討NKG2D受體和其相關配體ULBP在γδT細胞抑制胰腺癌裸鼠移植瘤生長中的作用。檢測胰腺癌組織中的ULBP表達。人體外周血中ULBP的含量對胰腺癌診斷價值。【方法】利用抗體吸附法從健康志愿者外周血中分離培養(yǎng)出γδT細胞。將人胰腺癌細胞株PANC-1注入BALB/C裸鼠背部皮下以建立裸鼠胰腺癌動物模型。把裸鼠隨機地分成吉西他濱治療組,對照組和γδT細胞組。體外分離培養(yǎng)γδT細胞,擴增后經(jīng)裸鼠尾靜脈注入。每周進行2次裸鼠重量和腫瘤體積的測量。隨后手術摘除腫瘤組織并計算相對腫瘤體積和腫瘤抑制率。運用WB和qPCR檢測ULBPs在三組裸鼠胰腺癌移植瘤中的表達。IHC檢測30個胰腺癌患者腫瘤組織的ULBPs的表達情況。ELISA法檢測胰腺癌患者及健康志愿者的外周血中ULBPs和CA199的表達水平及外周血中ULBPs對胰腺癌診斷價值。【結果】胰腺癌腫瘤生長在γδT細胞治療組受到明顯抑制(P=0.003)。并且其抑瘤率和吉西他濱治療組的腫瘤抑瘤率無明顯差別。人胰腺癌組織的IHC實驗及裸鼠瘤組織的WB實驗都顯示ULBP在胰腺癌組織中過表達。此外,WB實驗表明經(jīng)過γδT的治療后,ULBP-1和ULBP-3的表達水平會下降(P=0.025,P=0.019)�？扇苄訳LBP2在胰腺癌患者外周血中的表達要比健康人群外周血中的含量高;可溶性ULBP2有助于CA19-9對胰腺癌的診斷[AUC=0.88(95%CI:0.807~0.943);AUC=0.84(95%CI:0.761~0.917)]�！窘Y論】γδT細胞對胰腺癌生長增值具有顯著的抑制作用,其發(fā)揮作用的機制可能與ULBP異常表達相關�？扇苄訳LBP2聯(lián)合CA199診斷胰腺癌的效果優(yōu)于單獨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 緯 未 T cells. The expression of ULBP in pancreatic carcinoma was detected. [methods] 緯 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 緯 未 T cell group. 緯 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 緯 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 緯 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] 緯 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.
【學位授予單位】：安徽醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R735.9
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本文編號：2311373