本文選題：胰腺癌 + 預(yù)后　； 參考：《中國人民解放軍醫(yī)學(xué)院》2017年博士論文

【摘要】：研究背景及目的:胰腺癌惡性程度高,診斷困難,在目前中國惡性腫瘤致死率中排名第九,臨床表現(xiàn)為快速的發(fā)展和轉(zhuǎn)移。外科手術(shù)依然是胰腺癌的治療標(biāo)準(zhǔn),但多數(shù)患者在發(fā)現(xiàn)時已經(jīng)因為遠(yuǎn)處轉(zhuǎn)移和局部晚期腫瘤而成為手術(shù)禁忌。胰腺癌患者的五年生存率不足5%,即使經(jīng)過根治性手術(shù)切除以后的患者,其五年生存率仍不足20%。腫瘤微環(huán)境中的免疫抑制是腫瘤生長,轉(zhuǎn)移的關(guān)鍵因素,甚至可以誘導(dǎo)腫瘤的免疫逃逸。吲哚胺2,3-雙加氧酶(IDO)是色氨酸經(jīng)犬尿氨酸途徑代謝的限速酶,具有免疫抑制效應(yīng),在多種腫瘤中表達(dá)并與預(yù)后相關(guān)。其在胰腺癌中的表達(dá)和作用目前仍沒有報道。為研究IDO在胰腺癌中的預(yù)后意義以及可能的免疫治療潛能,我們利用臨床研究和基礎(chǔ)實驗相結(jié)合的方法分析IDO在胰腺癌發(fā)生發(fā)展中的作用和IDO相關(guān)免疫治療的效果。方法:(1)采用Western Blot法和免疫組織化學(xué)法檢測胰腺癌組織中的IDO表達(dá)量,并進(jìn)一步分析IDO及臨床數(shù)據(jù)與胰腺癌患者術(shù)后生存率的關(guān)系。(2)采用Real time PCR法和Western Blot法檢測胰腺癌細(xì)胞系的IDO表達(dá)量和IFN-γ的調(diào)控作用,并分析體外條件下IDO抑制劑和AHR抑制劑的對胰腺癌細(xì)胞系的細(xì)胞毒性作用,為進(jìn)一步體內(nèi)動物實驗奠定基礎(chǔ)。(3)使用免疫健全小鼠構(gòu)建皮下種植瘤模型,使用IDO抑制劑1-MT并聯(lián)合免疫佐劑R848聯(lián)合應(yīng)用,觀察其對胰腺癌的治療效果,并使用流式細(xì)胞術(shù)分析腫瘤中免疫細(xì)胞數(shù)量。(4)使用抗體刪除動物體內(nèi)CD8+T細(xì)胞,并使用AHR抑制劑阻斷AHR信號通路功能,觀察各組腫瘤生長情況,評價其效果。分析1-MT聯(lián)合R848治療過程中腫瘤引流淋巴結(jié)中DC細(xì)胞的變化情況。結(jié)果:(1) IDO在胰腺癌中有表達(dá),與腫瘤分化程度(P=0.017),TNM分期(P=0.03)和淋巴結(jié)轉(zhuǎn)移情況(P=0.04)相關(guān),而與年齡(P=0.87),性別(P=0.89),腫瘤大小(P=0.95),腫瘤位置(P=0.08)無關(guān)。并且胰腺癌組織中IDO的表達(dá)量(HR = 2.38 P =0.02)及淋巴結(jié)轉(zhuǎn)移情況(HR = 2.86 P = 0.01)與胰腺癌患者預(yù)后相關(guān),可以作為評價胰腺癌預(yù)后的獨立指標(biāo)。(2) IDO在胰腺癌細(xì)胞系中的表達(dá),并且IFN-γ能夠明顯上調(diào)IDO的表達(dá)量;同時進(jìn)行了 1-MT、CH223191、R848的體外細(xì)胞毒性實驗,三種藥物在體外條件下對細(xì)胞增殖無明顯影響,可以用于胰腺癌動物模型免疫治療相關(guān)實驗。(3)建立胰腺癌皮下種植瘤模型,觀察1-MT聯(lián)合R848治療胰腺癌效果,結(jié)果顯示1-MT聯(lián)合R848治療組有較好的抑制腫瘤生長的效果,而單獨使用同樣劑量的1-MT和R848并不能很好的控制腫瘤的生長,進(jìn)一步分析腫瘤中的免疫細(xì)胞,1-MT聯(lián)合R848治療組與對照組和R848組相比較,腫瘤中CD8+T細(xì)胞數(shù)目明顯升高。(4) 1-MT和R848聯(lián)合治療組cDC細(xì)胞數(shù)目明顯升高;CD8+T細(xì)胞刪除組和CH223191+R848組,并不能抑制腫瘤生長。結(jié)論:胰腺癌組織中IDO的表達(dá)量與胰腺癌患者預(yù)后相關(guān),并且IDO可以作為評價胰腺癌預(yù)后的獨立指標(biāo);IDO在胰腺癌中的表達(dá)量受IFN- γ調(diào)控;1-MT和R848聯(lián)合治療抑制胰腺癌的效應(yīng)是通過cDC提呈腫瘤抗原,活化T細(xì)胞,增加腫瘤中浸潤C(jī)D8+T細(xì)胞來實現(xiàn)的,抑制IDO下游的AHR通路,并不能抑制腫瘤生長。
[Abstract]:Background and objective: pancreatic cancer is highly malignant and difficult to diagnose. It ranks ninth in the mortality rate of malignant tumor in China. The clinical manifestation is rapid development and metastasis. Surgery is still the standard of treatment for pancreatic cancer, but most of the patients have become contraindications to the operation because of distant metastasis and local advanced tumor. The five year survival rate of cancer patients is less than 5%. Even after radical resection, the five-year survival rate is still less than that of 20%. tumor microenvironment. The key factor of metastasis can induce tumor immune escape. The indolamine 2,3- dioxygenase (IDO) is the tryptophan through the canine urinary ammonia pathway. The rate limiting enzyme, which has the effect of immunosuppression, is expressed in a variety of tumors and is related to the prognosis. Its expression and role in pancreatic cancer are still not reported. To study the prognostic significance and potential immunotherapy potential of IDO in pancreatic cancer, we used a combination of clinical and basic testing to analyze the incidence of IDO in pancreatic cancer. The effect of the exhibition and the effect of IDO related immunotherapy. Methods: (1) the expression of IDO in pancreatic cancer tissues was detected by Western Blot and immunohistochemistry, and the relationship between IDO and clinical data and postoperative survival rate of pancreatic cancer patients was further analyzed. (2) the IDO table of pancreatic cancer cell lines was detected by Real time PCR method and Western Blot method. The effects of dose and IFN- gamma on the cytotoxicity of IDO inhibitors and AHR inhibitors on pancreatic cancer cell lines in vitro were analyzed. (3) the model of subcutaneous implants was constructed in immunized mice, combined with IDO inhibitor 1-MT and combined with immuno adjuvant R848. The therapeutic effect of pancreatic cancer and the use of flow cytometry to analyze the number of immune cells in the tumor. (4) the use of antibodies to delete CD8+T cells in animals, and the use of AHR inhibitors to block the function of AHR signal pathway, observe the growth of the tumor and evaluate the effect of the tumor, and analyze the changes of DC cells in the tumor drainage lymph nodes in the 1-MT combined with R848 treatment. Results: (1) IDO was expressed in pancreatic cancer, associated with tumor differentiation (P=0.017), TNM staging (P=0.03) and lymph node metastasis (P=0.04), but not related to age (P=0.87), sex (P=0.89), tumor size (P=0.95), and tumor location (P= 0.08). And the expression of IDO in pancreatic cancer tissues (HR = 2.38 P) and lymph node metastasis The condition (HR = 2.86 P = 0.01) is associated with the prognosis of pancreatic cancer patients. (2) the expression of IDO in the pancreatic cancer cell line, and IFN- gamma can obviously increase the expression of IDO; meanwhile, the cytotoxicity test of 1-MT, CH223191, R848 in vitro is carried out, and the three drugs can proliferate in vitro. No obvious influence, can be used in the animal model of pancreatic cancer immunotherapy related experiments. (3) to establish a model of pancreatic cancer subcutaneous implants and observe the effect of 1-MT combined with R848 in the treatment of pancreatic cancer. The results show that the combination of 1-MT and R848 has a better effect on inhibiting the growth of the tumor, and the same dose of 1-MT and R848 alone can not be well controlled. The tumor growth, further analysis of the tumor immune cells, 1-MT combined with R848 treatment group and the control group and the R848 group compared with the R848 group, the number of CD8+T cells increased significantly. (4) the number of cDC cells in the 1-MT and R848 combined treatment group increased significantly; CD8+T cell deletion group and CH223191+R848 group did not inhibit the tumor growth. Conclusion: I in the pancreatic cancer tissue, I. The expression of DO is associated with the prognosis of patients with pancreatic cancer, and IDO can be used as an independent index for evaluating the prognosis of pancreatic cancer. The expression of IDO in pancreatic cancer is regulated by IFN- gamma. The effect of 1-MT and R848 in the treatment of pancreatic cancer is achieved by presenting tumor antigen by cDC, activating T cells and adding CD8+T cells in the tumor to suppress IDO. The AHR pathway of swimming does not inhibit the growth of the tumor.
【學(xué)位授予單位】：中國人民解放軍醫(yī)學(xué)院
【學(xué)位級別】：博士
【學(xué)位授予年份】：2017
【分類號】：R735.9

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