A new liquid chromatography mass spectrometry method for the detection of paracrine low abundance proteome in tumor microenvironment

doi:10.3969/j.issn.1009-976X.2022.03.005

Abstract

Abstract: Objective A new liquid chromatography mass spectrometry method was established to detect paracrine low abundance proteins in tumor microenvironment. Methods Proteins of C57/B6 mouse subcutaneous fat primary cell culture medium were extracted with acetone, and analyzed by four-dimensional high field asymmetric waveform ion mobility spectrum electrostatic field orbital trap linear ion trap mode （4D-Faims-OTIT）, electron transfer cracking electrostatic field orbital trap linear ion trap mode （ETD-OTIT） and electrostatic field orbital trap linear ion trap mode （OTIT）. Results The protein identification numbers of 4D-Faims-OTIT, OTIT and ETD-OTIT in S1 samples were 1040±21, 428±4 and 274±6, respectively, and those in S2 samples were 851±31, 290±10 and 192±14, respectively. The modification sites in S1 were 138±12, 105±1 and 75±14, respectively. In S2 sample, they were 122±15, 66±15 and 42±7, respectively. The secretory proteins of S1 sample identified in 4D-Faims-OTIT were GDF15, CXCL12, Cathepsin D/B, Insulin-like growth factor and Apolipoprotein E/Cystatin C in colon cancer, lung cancer, pancreatic cancer, ovarian cancer and liver cancer, respectively. Conclusion 4D-Faims-OTIT can significantly improve the detection sensitivity of adipocyte secretory proteome in tumor microenvironment, and contribute to the study of the mechanism of surgical diseases such as colorectal cancer, lung cancer and ovarian cancer.

Key words: tumor microenvironment, paracrine low abundance protein, liquid chromatography mass spectrometry, 4D-Faims-OTIT, ETD-OTIT, OTIT

摘要： 目的建立一种新液质联用方法检测肿瘤微环境中旁分泌型低丰度蛋白。方法用丙酮提取C57/B6小鼠皮下脂肪原代细胞培养基中的蛋白质,通过四维-高场非对称波形离子迁移谱-静电场轨道阱线性离子阱模式（4D-Faims-OTIT）,电子转移裂解-静电场轨道阱线性离子阱模式（ETD-OTIT）和静电场轨道阱线性离子阱模式（OTIT）三种液质联用模式分析。结果 S1样本4D-Faims-OTIT,OTIT和ETD的蛋白鉴定数分别为1040±21,428±4和274±6,S2样本分别为851±31,290±10和192±14。S1中修饰位点分别为138±12,105±1和75±14,S2中分别为122±15,66±15和42±7。4D-Faims-OTIT在S1中鉴定到分泌型蛋白为结肠癌（GDF15）、肺癌（CXCL12）、胰腺癌（Cathepsin D/B）、卵巢癌（Insulin-like growth factor）、肝癌（Apolipoprotein E/ Cystatin C）。结论 4D-Faims-OTIT可显著提高肿瘤微环境中脂肪细胞分泌型蛋白组的检测灵敏度,有助于结直肠癌、肺癌和卵巢癌等疾病机理的研究。

关键词: 肿瘤微环境, 旁分泌型低丰度蛋白, 液质联用, 4D-Faims-OTIT, ETD-OTIT, OTIT

CLC Number:

R73-3

WANG Xiao-juan, HUANG Yu-na, YAO Wei-cheng, FU Yu, CEN Mei-feng. A new liquid chromatography mass spectrometry method for the detection of paracrine low abundance proteome in tumor microenvironment[J]. Lingnan Modern Clinics In Surgery, 2022, 22(03): 236-244.

王小娟, 黄雨娜, 姚伟城, 浮钰, 岑美凤. 肿瘤微环境的旁分泌型低丰度蛋白组检测的新液质联用方法[J]. 岭南现代临床外科, 2022, 22(03): 236-244.

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