胰腺癌不同进展期血清外泌体蛋白质组学分析

doi:10.6040/j.issn.1671-7554.0.2022.0169

摘要/Abstract

摘要： 目的筛选健康对照与不同进展期胰腺癌患者的血清外泌体差异蛋白质,绘制差异蛋白质表达图谱,并分析差异蛋白质参与的生物学过程。方法采用非标记定量蛋白质组学进行外泌体蛋白质检测,筛选差异蛋白质并进行生物信息学分析;利用蛋白免疫印迹对与胰腺癌进展相关的蛋白质进行血清学验证。结果在对照组、胰腺癌未转移组和转移组样品中共检测到2 242个蛋白质。通过不同分组差异蛋白质分析,共鉴定出664个差异具有统计学意义的蛋白质,主要参与囊泡介导转运、局灶黏附、细胞外基质(ECM)受体相互作用等信号通路。此外,鉴定到在3组间差异均有统计学意义的蛋白质5个,验证发现富亮氨酸α2糖蛋白1(LRG1)在胰腺癌转移组外泌体中表达量较高。结论对照组、胰腺癌未转移组和转移组间蛋白质表达存在差异,差异蛋白质主要富集于局灶黏附、细胞外基质受体相互作用等信号通路,且在3组间表达均有差异的外泌体蛋白质LRG1可能成为监测胰腺癌进展的血清学标志物。

关键词: 胰腺癌, 外泌体, 蛋白质组学, 生物信息学, 富亮氨酸α2糖蛋白1

Abstract: Objective To screen the differential proteins of serum exosomes in healthy controls and pancreatic cancer patients with different degrees of progression, draw the profile, and analyze their functions and biological processes. Methods Exosomes were quantitatively analyzed with label-free quantitative proteomic analysis technology. The differential proteins were screened and bioinformatic analysis was performed. The proteins associated with pancreatic cancer progression were serologically verified with Western blotting. Results A total of 2 242 proteins were detected in the control, non-metastatic and metastatic pancreatic cancer samples with label-free quantitative proteomic analysis and a total of 664 proteins with statistically significant differences were identified, which were mainly involved in vesicle-mediated transport, focal adhesion and extracellular cortex matrix(ECM)receptor interaction. Moreover, there were 5 proteins with statistically significant differences among the three groups, of which leucine-rich alpha-2-glycoprotein 1(LRG1)was highly expressed in serum exosomes of metastatic pancreatic cancer. Conclusion There are differences in protein expression patterns among healthy controls, non-metastatic and metastatic pancreatic cancer patients, and the differential proteins are mainly enriched in focal adhesion and ECM-receptor interaction and other signaling pathways. LRG1, which is different in all three groups, may be a serological marker for monitoring the progression of pancreatic cancer.

Key words: Pancreatic cancer, Exosome, Proteomics, Bioinformatics, Leucine-rich alpha-2-glycoprotein 1

中图分类号:

R735.9

李雁儒,李娟,李培龙,杜鲁涛,王传新. 胰腺癌不同进展期血清外泌体蛋白质组学分析[J]. 山东大学学报 (医学版), 2022, 60(10): 33-41.

LI Yanru, LI Juan, LI Peilong, DU Lutao, WANG Chuanxin. Proteomic analysis of serum exosomes in pancreatic cancer with different stages of progress[J]. Journal of Shandong University (Health Sciences), 2022, 60(10): 33-41.

参考文献

[1] Klein AP. Pancreatic cancer epidemiology: understanding the role of lifestyle and inherited risk factors[J]. Nat Rev Gastroenterol Hepatol, 2021, 18(7):493-502.
[2] Siegel RL, Miller KD, Fuchs HE, et al. Cancer statistics, 2021[J]. CA Cancer J Clin, 2021, 71(1): 7-33.
[3] Vincent A, Herman J, Schulick R, et al. Pancreatic cancer[J]. Lancet, 2011, 378(9791): 607-620.
[4] Jiang K, Chen H, Fang Y, et al. Exosomal ANGPTL1 attenuates colorectal cancer liver metastasis by regulating Kupffer cell secretion pattern and impeding MMP9 induced vascular leakiness[J]. J Exp Clin Cancer Res, 2021, 40(1): 21.
[5] Chen G, Huang AC, Zhang W, et al. Exosomal PD-L1 contributes to immunosuppression and is associated with anti-PD-1 response[J]. Nature, 2018, 560(7718): 382-386.
[6] 韩柳, 孙宇. 胞外囊泡与癌症[J]. 中国细胞生物学学报, 2016, 38(4): 347-355. HAN Liu, SUN Yu. Extracellular vesicles and cancer[J]. Chinese Journal of Cell Biology, 2016, 38(4): 347-355.
[7] Melo SA, Luecke LB, Kahlert C, et al. Glypican-1 identifies cancer exosomes and detects early pancreatic cancer[J]. Nature, 2015, 523(7559): 177-182.
[8] Liu Z, Liu Y, Qian L, et al. A proteomic and phosphoproteomic landscape of KRAS mutant cancers identifies combination therapies[J]. Mol Cell, 2021, 81(19): 4076-4090.
[9] Liu J, Yuan B, Cao J, et al. Ambra1 promotes TGFbeta signaling via nonproteolytic polyubiquitylation of smad4[J]. Cancer Res, 2021, 81(19): 5007-5020.
[10] Wu J, Gao W, Tang Q, et al. M2 macrophage-derived exosomes facilitate HCC metastasis by transferring alphaM beta2 integrin to tumor cells[J]. Hepatology, 2021, 73(4): 1365-1380.
[11] Wang D, Zhao C, Xu F, et al. Cisplatin-resistant NSCLC cells induced by hypoxia transmit resistance to sensitive cells through exosomal PKM2[J]. Theranostics, 2021, 11(6): 2860-2875.
[12] Yuan X, Qian N, Ling S, et al. Breast cancer exosomes contribute to pre-metastatic niche formation and promote bone metastasis of tumor cells[J]. Theranostics, 2021, 11(3): 1429-1445.
[13] Wang S, Xu M, Li X, et al. Exosomes released by hepatocarcinoma cells endow adipocytes with tumor-promoting properties[J]. J Hematol Oncol, 2018, 11(1): 82.
[14] Yoon JH, Ashktorab H, Smoot DT, et al. Uptake and tumor-suppressive pathways of exosome-associated GKN1 protein in gastric epithelial cells[J]. Gastric Cancer, 2020, 23(5): 848-862.
[15] Repetto O, De Re V, Giuffrida P, et al. Proteomics signature of autoimmune atrophic gastritis: towards a link with gastric cancer[J]. Gastric Cancer, 2021, 24(3): 666-679.
[16] Thakur R, Singh PK. Molecular subtypes of pancreatic cancer: a proteomics approach[J]. Clin Cancer Res, 2021, 27(12): 3272-3274.
[17] Kugeratski FG, Hodge K, Lilla S, et al. Quantitative proteomics identifies the core proteome of exosomes with syntenin-1 as the highest abundant protein and a putative universal biomarker[J]. Nat Cell Biol, 2021, 23(6): 631-641.
[18] Yang J, Zhang Y, Gao X, et al. Plasma-derived exosomal ALIX as a novel biomarker for diagnosis and classification of pancreatic cancer[J]. Front Oncol, 2021, 11: 628346. doi: 10.3389/fonc.2021.628346.
[19] Han S, Huo Z, Nguyen K, et al. The proteome of pancreatic cancer-derived exosomes reveals signatures rich in key signaling pathways[J]. Proteomics, 2019, 19(13): e1800394. doi: 10.1002/pmic.201800394.
[20] Castillo J, Bernard V, San Lucas FA, et al. Surfaceome profiling enables isolation of cancer-specific exosomal cargo in liquid biopsies from pancreatic cancer patients[J]. Ann Oncol, 2018, 29(1): 223-229.
[21] Shi X, Guo X, Li X, et al. Loss of Linc01060 induces pancreatic cancer progression through vinculin-mediated focal adhesion turnover[J]. Cancer Lett, 2018, 433: 76-85. doi: 10.1016/j.canlet.2018.06.015.
[22] Okamoto H, Kusama T, Fujii H. Tyrosine phosphorylation of focal adhesion anchoring proteins enhances human pancreatic cancer cell invasion[J]. Pancreas, 2016, 45(7): 37-39.
[23] Wang X, Luo G, Zhang K, et al. Hypoxic tumor-derived exosomal miR-301a mediates M2 macrophage polarization via PTEN/PI3Kgamma to promote pancreatic cancer metastasis[J]. Cancer Res, 2018, 78(16): 4586-4598.
[24] Yang B, Feng X, Liu H, et al. High-metastatic cancer cells derived exosomal miR92a-3p promotes epithelial-mesenchymal transition and metastasis of low-metastatic cancer cells by regulating PTEN/Akt pathway in hepatocellular carcinoma[J]. Oncogene, 2020, 39(42): 6529-6543.
[25] Chen Q, Yu D, Zhao Y, et al. Screening and identification of hub genes in pancreatic cancer by integrated bioinformatics analysis[J]. J Cell Biochem, 2019, 120(12): 19496-19508.
[26] Chen S, Gao C, Yu T, et al. Bioinformatics analysis of a prognostic miRNA signature and potential key genes in pancreatic cancer[J]. Front Oncol, 2021, 11: 641289. doi: 10.3389/fonc.2021.641289.
[27] Singhal M, Gengenbacher N, Abdul Pari AA, et al. Temporal multi-omics identifies LRG1 as a vascular niche instructor of metastasis[J]. Sci Transl Med, 2021, 13(609): eabe6805. doi: 10.1126/scitranslmed.abe6805.
[28] He L, Feng A, Guo H, et al. LRG1 mediated by ATF3 promotes growth and angiogenesis of gastric cancer by regulating the SRC/STAT3/VEGFA pathway[J]. Gastric Cancer, 2022, 25(3): 527-541.
[29] Zhong B, Cheng B, Huang X, et al. Colorectal cancer-associated fibroblasts promote metastasis by up-regulating LRG1 through stromal IL-6/STAT3 signaling[J]. Cell Death Dis, 2021, 13(1): 16. doi: 10.1038/s41419-021-04461-6.

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