Versican影响胰腺神经内分泌瘤预后的作用机制及晚期进展性胰腺神经内分泌瘤的临床资料分析

发布时间：2018-05-15 18:00

  本文选题：胰腺神经内分泌瘤 + Versican　； 参考：《北京协和医学院》2016年博士论文

【摘要】：[背景]胰腺神经内分泌瘤(pancreatic neuroendocrine tumor, pNE T)为罕见肿瘤,但其发病率逐年上升,相同分期的pNET患者预后差异很大。因此寻找与预后相关的特异性分子标志物,指导临床治疗和判断预后是目前pNET研究的热点和重点。Versican (VCAN)是一种大分子硫酸软骨素蛋白多糖,我们前期的蛋白质芯片研究结果显示Versican在pNET肿瘤组织中表达阳性率高,本研究拟扩大样本量检测Versican在pNET组织中的表达水平,回顾性分析Versican与pNET患者临床预后的关系。[方法]回顾性收集2005至2010年间的127例pNET患者的完整临床病理资料及随访结果。应用免疫组织化学染色检测肿瘤组织及瘤旁组织中的Versican蛋白表达水平,探讨pNET肿瘤组织与瘤旁组织间Versican蛋白表达阳性率有无差异；结合患者信息,分析pNET肿瘤组织中Versican表达阳性率与pNET患者的临床病理特点及预后的相关性。[结果]127例pNET肿瘤组织及46例瘤旁组织Versican免疫组化染色结果显示,46例瘤旁组织中的胰岛细胞表达均为阳性；肿瘤组织中52例表达阴性(40.9%),75例表达阳性(59.1%)。肿瘤小于2cm、肿瘤分期Ⅰ-Ⅱ期及胰岛素瘤的pNET患者中Versican的表达阳性率高于肿瘤大于2cm、分期为Ⅲ-Ⅳ期或非胰岛素瘤的患者(p0.05)。单因素生存分析显示肿瘤组织中Versican表达阳性患者的无病生存期(disease-free survival, DF S)有延长的趋势,结果接近有统计学意义(p=0.08)。亚组分析中,G2患者亚组、NO亚组和F-pNET亚组中,Versican表达阳性患者的DFS与Versican表达阴性患者相比有延长的趋势,结果接近统计学意义(分别为p=0.056,p=0.072,p=0.061)。多因素分析显示肿瘤分期与肿瘤分级是pNET患者的独立预后因素。[结论]在肿瘤组织中Versican表达阳性率与肿瘤大小、肿瘤分期、是否为胰岛素瘤存在显著相关性；肿瘤组织中Versican日性表达与患者预后可能相关。[背景]Versican(VCAN)是一种大分子硫酸软骨素蛋白多糖,我们前期研究发现Versican在pNET肿瘤组织中表达阳性率高,并且Versican表达阳性的pNET患者的无病生存期更长；但Versican在pNET中的作用机制尚不清楚。本部分进一步对Versican在pNET细胞系中的作用机制进行研究。[方法]首先利用定量PCR检测Versican不同亚型在pNET肿瘤组织和人源pNET细胞系(BON-1)中的表达水平；然后设计Versican的干扰序列,利用脂质体转染法在BON-1中分别转染Versican-siRNA及相应对照,采用流式细胞术及定量PCR法(qRT-PCR)检测转染效率。然后检测Versican干扰后对BON-1细胞生物学功能的影响。利用MT S法检测各组细胞增殖；流式细胞术分析各组细胞周期及细胞凋亡; Transwell小室分析各组细胞迁移。采用Western blot及qRT-PCR法检测Versican相关信号通路的基因表达。[结果]Versican VO亚型在pNET组织及pNET细胞系中的表达水平均明显高于其他三种亚型。在BON-1细胞系中,Versican干扰组与其对照相比,细胞的迁移无统计学差异(p0.05); Versican干扰后BON-1细胞的增殖率明显增高,同时细胞周期显示处于G2/M期的细胞百分比明显升高(p0.05)：定量PCR法及Western Blot法发现Versican干扰后p27的表达水平较对照组显著下降(p0.05),而CDK2表达水平较对照组显著上升(p0.05)。[结论]本研究发现Versican VO亚型是人pNET的主要表达亚型。Versican表达下调可能通过p27/CDK2信号通路,导致pNET细胞进入G2/M期的细胞增多,从而促进pNET细胞增殖。[背景]药物治疗成为晚期进展的胰腺神经内分泌瘤(pancreatic neuroendocrine tumor, pNET)的主要治疗方式。药物选择种类有很多,包括生长抑素类似物、靶向药物(舒尼替尼、依维莫司)和化疗药物等。但国人晚期pNET患者应用各种治疗的疗效尚无比较数据,本研究旨在回顾性分析药物治疗对中国晚期进展的pNET患者的临床疗效。[方法]收集北京协和医院肿瘤内科药物治疗的晚期进展的pNET患者的临床病理资料、方案及生存预后等信息。使用药物包括生长抑素类似物(somatostatin analogs, SSA),靶向药物(舒尼替尼、依维莫司、索凡替尼)和化疗药物(替莫唑胺、达卡巴嗪、氟脲嘧啶类)。[结果]共55例pNET患者纳入研究。全部患者的5年和10年总生存率分别为75%和62.5%。一线应用SSA治疗的患者(29例)与应用靶向治疗患者(15例)的中位PFS (median PFS, mPFS)比没有统计学差异(7个月和12个月,p0.05)；二线应用联合方案治疗的患者(10例)与靶向治疗患者(12例)的mPFS相比没有统计学差异(10个月和7个月,p0.05)。在SSA治疗亚组,Ki-67 10-20%患者(10例)的mPFS (5个月)较Ki-6710%患者(19例)的mPFS(9个月)短,结果没有统计学意义(P=0.2)。在Ki-6710-20%的患者亚组,应用化疗患者(9例)的mPFS(12个月)长于应用SSA患者(10例)的mPFS(5个月),结果没有统计学意义(P=0.16)。一线应用舒尼替尼治疗的患者9例,二线及二线以上应用舒尼替尼治疗的患者9例,两组的mPFS均为12m,1年-PFS%, ORR和DCR分别为44.4%,22.2%,88.9%和43.7%,33.3%,77.8%,结果无明显差异(P0.05)。Ki-6710%患者应用舒尼替尼的mPFS(13m)长于Ki-67≥10%患者(8m),结果无统计学差异(P0.05)。舒尼替尼的主要不良反应为骨髓抑制(14/17,82.4%)、腹泻(9/17,52.9%)、高血压(8/17,47.1%)、蛋白尿(6/17,35.3%)和皮疹(6/17,35.3%)。[结论]在晚期进展的pNET患者中,一线应用SSA治疗与靶向治疗的疗效无明显差异,但靶向治疗有PFS延长的趋势；Ki-67可能预示药物治疗的疗效。舒尼替尼治疗中国晚期进展pNET的疗效和不良反应与西方数据相似,一线应用和二线及二线以上应用的疗效可能无差异。我们仍需扩大样本量探讨pNET患者对各种药物治疗的最佳获益。
[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.

【学位授予单位】：北京协和医学院
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R735.9

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