本文選題：胰腺神經(jīng)內(nèi)分泌瘤 + Versican��； 參考：《北京協(xié)和醫(yī)學(xué)院》2016年博士論文

【摘要】：[背景]胰腺神經(jīng)內(nèi)分泌瘤(pancreatic neuroendocrine tumor, pNE T)為罕見腫瘤,但其發(fā)病率逐年上升,相同分期的pNET患者預(yù)后差異很大。因此尋找與預(yù)后相關(guān)的特異性分子標(biāo)志物,指導(dǎo)臨床治療和判斷預(yù)后是目前pNET研究的熱點(diǎn)和重點(diǎn)。Versican (VCAN)是一種大分子硫酸軟骨素蛋白多糖,我們前期的蛋白質(zhì)芯片研究結(jié)果顯示Versican在pNET腫瘤組織中表達(dá)陽性率高,本研究擬擴(kuò)大樣本量檢測Versican在pNET組織中的表達(dá)水平,回顧性分析Versican與pNET患者臨床預(yù)后的關(guān)系。[方法]回顧性收集2005至2010年間的127例pNET患者的完整臨床病理資料及隨訪結(jié)果。應(yīng)用免疫組織化學(xué)染色檢測腫瘤組織及瘤旁組織中的Versican蛋白表達(dá)水平,探討pNET腫瘤組織與瘤旁組織間Versican蛋白表達(dá)陽性率有無差異；結(jié)合患者信息,分析pNET腫瘤組織中Versican表達(dá)陽性率與pNET患者的臨床病理特點(diǎn)及預(yù)后的相關(guān)性。[結(jié)果]127例pNET腫瘤組織及46例瘤旁組織Versican免疫組化染色結(jié)果顯示,46例瘤旁組織中的胰島細(xì)胞表達(dá)均為陽性；腫瘤組織中52例表達(dá)陰性(40.9%),75例表達(dá)陽性(59.1%)。腫瘤小于2cm、腫瘤分期Ⅰ-Ⅱ期及胰島素瘤的pNET患者中Versican的表達(dá)陽性率高于腫瘤大于2cm、分期為Ⅲ-Ⅳ期或非胰島素瘤的患者(p0.05)。單因素生存分析顯示腫瘤組織中Versican表達(dá)陽性患者的無病生存期(disease-free survival, DF S)有延長的趨勢,結(jié)果接近有統(tǒng)計(jì)學(xué)意義(p=0.08)。亞組分析中,G2患者亞組、NO亞組和F-pNET亞組中,Versican表達(dá)陽性患者的DFS與Versican表達(dá)陰性患者相比有延長的趨勢,結(jié)果接近統(tǒng)計(jì)學(xué)意義(分別為p=0.056,p=0.072,p=0.061)。多因素分析顯示腫瘤分期與腫瘤分級(jí)是pNET患者的獨(dú)立預(yù)后因素。[結(jié)論]在腫瘤組織中Versican表達(dá)陽性率與腫瘤大小、腫瘤分期、是否為胰島素瘤存在顯著相關(guān)性；腫瘤組織中Versican日性表達(dá)與患者預(yù)后可能相關(guān)。[背景]Versican(VCAN)是一種大分子硫酸軟骨素蛋白多糖,我們前期研究發(fā)現(xiàn)Versican在pNET腫瘤組織中表達(dá)陽性率高,并且Versican表達(dá)陽性的pNET患者的無病生存期更長；但Versican在pNET中的作用機(jī)制尚不清楚。本部分進(jìn)一步對(duì)Versican在pNET細(xì)胞系中的作用機(jī)制進(jìn)行研究。[方法]首先利用定量PCR檢測Versican不同亞型在pNET腫瘤組織和人源pNET細(xì)胞系(BON-1)中的表達(dá)水平；然后設(shè)計(jì)Versican的干擾序列,利用脂質(zhì)體轉(zhuǎn)染法在BON-1中分別轉(zhuǎn)染Versican-siRNA及相應(yīng)對(duì)照,采用流式細(xì)胞術(shù)及定量PCR法(qRT-PCR)檢測轉(zhuǎn)染效率。然后檢測Versican干擾后對(duì)BON-1細(xì)胞生物學(xué)功能的影響。利用MT S法檢測各組細(xì)胞增殖；流式細(xì)胞術(shù)分析各組細(xì)胞周期及細(xì)胞凋亡; Transwell小室分析各組細(xì)胞遷移。采用Western blot及qRT-PCR法檢測Versican相關(guān)信號(hào)通路的基因表達(dá)。[結(jié)果]Versican VO亞型在pNET組織及pNET細(xì)胞系中的表達(dá)水平均明顯高于其他三種亞型。在BON-1細(xì)胞系中,Versican干擾組與其對(duì)照相比,細(xì)胞的遷移無統(tǒng)計(jì)學(xué)差異(p0.05); Versican干擾后BON-1細(xì)胞的增殖率明顯增高,同時(shí)細(xì)胞周期顯示處于G2/M期的細(xì)胞百分比明顯升高(p0.05)：定量PCR法及Western Blot法發(fā)現(xiàn)Versican干擾后p27的表達(dá)水平較對(duì)照組顯著下降(p0.05),而CDK2表達(dá)水平較對(duì)照組顯著上升(p0.05)。[結(jié)論]本研究發(fā)現(xiàn)Versican VO亞型是人pNET的主要表達(dá)亞型。Versican表達(dá)下調(diào)可能通過p27/CDK2信號(hào)通路,導(dǎo)致pNET細(xì)胞進(jìn)入G2/M期的細(xì)胞增多,從而促進(jìn)pNET細(xì)胞增殖。[背景]藥物治療成為晚期進(jìn)展的胰腺神經(jīng)內(nèi)分泌瘤(pancreatic neuroendocrine tumor, pNET)的主要治療方式。藥物選擇種類有很多,包括生長抑素類似物、靶向藥物(舒尼替尼、依維莫司)和化療藥物等。但國人晚期pNET患者應(yīng)用各種治療的療效尚無比較數(shù)據(jù),本研究旨在回顧性分析藥物治療對(duì)中國晚期進(jìn)展的pNET患者的臨床療效。[方法]收集北京協(xié)和醫(yī)院腫瘤內(nèi)科藥物治療的晚期進(jìn)展的pNET患者的臨床病理資料、方案及生存預(yù)后等信息。使用藥物包括生長抑素類似物(somatostatin analogs, SSA),靶向藥物(舒尼替尼、依維莫司、索凡替尼)和化療藥物(替莫唑胺、達(dá)卡巴嗪、氟脲嘧啶類)。[結(jié)果]共55例pNET患者納入研究。全部患者的5年和10年總生存率分別為75%和62.5%。一線應(yīng)用SSA治療的患者(29例)與應(yīng)用靶向治療患者(15例)的中位PFS (median PFS, mPFS)比沒有統(tǒng)計(jì)學(xué)差異(7個(gè)月和12個(gè)月,p0.05)；二線應(yīng)用聯(lián)合方案治療的患者(10例)與靶向治療患者(12例)的mPFS相比沒有統(tǒng)計(jì)學(xué)差異(10個(gè)月和7個(gè)月,p0.05)。在SSA治療亞組,Ki-67 10-20%患者(10例)的mPFS (5個(gè)月)較Ki-6710%患者(19例)的mPFS(9個(gè)月)短,結(jié)果沒有統(tǒng)計(jì)學(xué)意義(P=0.2)。在Ki-6710-20%的患者亞組,應(yīng)用化療患者(9例)的mPFS(12個(gè)月)長于應(yīng)用SSA患者(10例)的mPFS(5個(gè)月),結(jié)果沒有統(tǒng)計(jì)學(xué)意義(P=0.16)。一線應(yīng)用舒尼替尼治療的患者9例,二線及二線以上應(yīng)用舒尼替尼治療的患者9例,兩組的mPFS均為12m,1年-PFS%, ORR和DCR分別為44.4%,22.2%,88.9%和43.7%,33.3%,77.8%,結(jié)果無明顯差異(P0.05)。Ki-6710%患者應(yīng)用舒尼替尼的mPFS(13m)長于Ki-67≥10%患者(8m),結(jié)果無統(tǒng)計(jì)學(xué)差異(P0.05)。舒尼替尼的主要不良反應(yīng)為骨髓抑制(14/17,82.4%)、腹瀉(9/17,52.9%)、高血壓(8/17,47.1%)、蛋白尿(6/17,35.3%)和皮疹(6/17,35.3%)。[結(jié)論]在晚期進(jìn)展的pNET患者中,一線應(yīng)用SSA治療與靶向治療的療效無明顯差異,但靶向治療有PFS延長的趨勢；Ki-67可能預(yù)示藥物治療的療效。舒尼替尼治療中國晚期進(jìn)展pNET的療效和不良反應(yīng)與西方數(shù)據(jù)相似,一線應(yīng)用和二線及二線以上應(yīng)用的療效可能無差異。我們?nèi)孕钄U(kuò)大樣本量探討pNET患者對(duì)各種藥物治療的最佳獲益。
[Abstract]:[background] pancreatic neuroendocrine tumor (pNE T) is a rare tumor, but its incidence is increasing year by year. The prognosis of pNET patients with the same stage is very different. Therefore, it is a hot spot and key.Versican (VCA) to find the specific molecular markers related to the prognosis, to guide the clinical treatment and to judge the prognosis in pNET. N) is a macromolecule chondroitin sulfate proteoglycan. Our previous protein chip research results show that the positive rate of Versican expression in pNET tumor tissue is high. This study intends to expand the sample size to detect the expression level of Versican in the pNET tissue, and review the relationship between the Versican and the clinical prognosis of pNET patients. [Methods] retrospective collection Complete clinicopathological data and follow-up results of 127 patients with pNET from 2005 to 2010. Immunohistochemical staining was used to detect the expression of Versican protein in tumor tissue and para tumor tissue, and to explore the difference in the positive rate of Versican protein expression between the pNET tumor tissue and the para tumor tissue; the pNET tumor group was analyzed in combination with the patient information. The correlation between the positive rate of Versican expression in the tissue and the clinicopathological features and prognosis of pNET patients. [results]127 cases pNET tumor tissue and 46 cases of Para Para tissue Versican immunohistochemical staining showed that the expression of islet cells in 46 cases of para tumor tissues were all positive; 52 cases in tumor tissues were negative (40.9%), 75 cases were positive (59.1%). The tumor was less than 2cm, the positive rate of Versican expression in the tumor stage I - II stage and the pNET patients with insulinoma was higher than that of the tumor larger than 2cm, and the patients with stage III - IV or non insulin tumor (P0.05). The univariate survival analysis showed that the disease free survival (disease-free survival, DF S) in the tumor tissues was prolonged (disease-free survival, DF S). The results were close to statistical significance (p=0.08). In subgroup analysis, the DFS positive patients in G2 subgroup, NO subgroup and F-pNET subgroup had an extended trend compared with Versican negative patients, and the results were close to statistical significance (p=0.056, p= 0.072, p=0.061). Multifactor analysis showed tumor staging and tumor grading. It is an independent prognostic factor for pNET patients. [Conclusion] there is a significant correlation between the positive rate of Versican expression in the tumor tissue and the size of the tumor, the stage of the tumor, or not, and the daily expression of Versican in the tumor may be related to the prognosis of the patients. [background]Versican (VCAN) is a macromolecule chondroitin sulfate proteoglycan, before us. The study found that the positive rate of Versican was high in pNET tumor tissues, and the period of disease free survival of pNET patients with Versican positive was longer, but the mechanism of Versican in pNET was still unclear. This part further studied the mechanism of Versican in pNET cell line. [method] first use quantitative PCR to detect Versican. The expression level of different subtypes in pNET tumor tissue and human pNET cell line (BON-1); then the Versican interference sequence was designed, Versican-siRNA and corresponding control were transfected in BON-1 by liposome transfection. Flow cytometry and quantitative PCR (qRT-PCR) were used to detect the transfection efficiency. Then BON-1 fines were detected after Versican interference. The effect of cell biological function. The cell proliferation was detected by MT S method; cell cycle and apoptosis of each group were analyzed by flow cytometry; cell migration in each group was analyzed by Transwell compartment analysis. The gene expression of Versican related signaling pathway was detected by Western blot and qRT-PCR. [results] Versican VO subtype was in pNET tissue and pNET cell line The expression level in the BON-1 cell line was significantly higher than that of the other three subtypes. In the Versican interference group, there was no significant difference in the cell migration of the Versican interference group (P0.05), and the proliferation rate of BON-1 cells increased significantly after the Versican interference, and the cell cycle showed a significant increase in the percentage of cells in the phase of the cell (P0.05): quantitative PCR method and Wes The expression level of p27 was significantly lower than that of the control group (P0.05) after Versican interference (P0.05), and the expression level of CDK2 was significantly higher than that of the control group (P0.05). [Conclusion] this study found that the VO subtype of Versican VO is the main expression of the subtype.Versican expression of the pNET, which may pass through the p27/ signaling pathway and lead to the cells entering the period of the Versican. Increase, thus promoting pNET cell proliferation. [background] drug therapy has become a major treatment for advanced pancreatic neuroendocrine tumor (pancreatic neuroendocrine tumor, pNET). There are many kinds of drug selection, including somatostatin analogues, targeted drugs (sulonitinib, imoimus), and chemotherapeutic drugs. But late pNET development in Chinese people. There is no comparative data for the use of various treatments. The purpose of this study is to review the clinical efficacy of drug therapy for pNET patients with advanced progress in China. [Methods] collect information on the clinicopathological data of pNET patients with advanced advances in the medical treatment of oncology in Peking Union Medical College Hospital. Somatostatin analogs (SSA), targeted drugs (sulonitinib, everimus, somatinib) and chemotherapeutic drugs (temozolomide, Dhaka basazine, fluorouracil). [results] a total of 55 patients with pNET were included in the study. The total survival rate of 5 and 10 years in all patients was 75% and SSA in the first line of 62.5%. (29 cases). The median PFS (median PFS, mPFS) in the target treatment (15 cases) had no statistical difference (7 months and 12 months, P0.05); the second line combined regimen (10 cases) had no statistical difference compared with the target therapy (12 cases) mPFS (10 months and 7 months, P0.05). In the SSA treatment subgroup, Ki-67 10-20% patients (10 cases) mPFS. (5 months) compared with mPFS (9 months) of Ki-6710% patients (19 cases), the results were not statistically significant (P=0.2). In the subgroup of Ki-6710-20% patients, the mPFS (12 months) of the patients with chemotherapy (12 months) was longer than the mPFS (5 months) of the SSA patients (10 cases). The results were not statistically significant (P=0.16). The first line used sulanitinib for 9 cases, second and second line. 9 patients were treated with suneinib. The mPFS in the two group was 12M, 1 years -PFS%, ORR and DCR were 44.4%, 22.2%, 88.9% and 43.7%, 33.3%, 77.8%. The results were no significant difference (13m) in.Ki-6710% patients (13m) longer than Ki-67 > 10% (8m). There was no significant difference (P0.05). Myelosuppression (14/17,82.4%), diarrhea (9/17,52.9%), hypertension (8/17,47.1%), proteinuria (6/17,35.3%) and rash (6/17,35.3%). [Conclusion] there is no significant difference in the efficacy of SSA therapy and targeting therapy in advanced advanced pNET patients, but targeting therapy has a tendency to prolong PFS, and Ki-67 may indicate the curative effect of drug therapy. The efficacy and adverse effects of nimatinib in the treatment of advanced pNET in China are similar to those of Western data. There may be no difference in the efficacy of first line applications and second line and above two line applications. We still need to expand the sample size to explore the best benefits of pNET patients for various drug treatments.

【學(xué)位授予單位】：北京協(xié)和醫(yī)學(xué)院
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：R735.9

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