您的位置:山东大学 -> 科技期刊社 -> 《山东大学学报(医学版)》

山东大学学报 (医学版) ›› 2021, Vol. 59 ›› Issue (9): 140-147.doi: 10.6040/j.issn.1671-7554.0.2021.1064

• 科研论著 • 上一篇    下一篇

下咽鳞癌蛋白质谱鉴定及预后靶分子筛选

陈立晓,英信江,陈歆维,王菲,孙臻峰,董频   

  • 发布日期:2021-10-15
  • 通讯作者: 董频. E-mail:dongpin64@aliyun.com
  • 基金资助:
    国家自然科学基金(82072989);上海市申康医院发展中心-新兴前沿技术联合攻关项目(SHDC12020120)

Identification of proteins in hypopharyngeal squamous cell carcinoma by mass spectrometry and screening of prognostic target molecules

CHEN Lixiao, YING Xinjiang, CHEN Xinwei, WANG Fei, SUN Zhenfeng, DONG Pin   

  1. Department of Otorhinolaryngology Head and Neck Surgery, Shanghai General Hospital, Shanghai 200080, China
  • Published:2021-10-15

摘要: 目的 利用非数据依赖性采集(DIA)蛋白质组学技术筛选下咽鳞癌晚期肿瘤预后相关的特异性靶分子。 方法 选取2016年1月至2018年9月在上海市第一人民医院确诊的5对晚期下咽鳞癌的癌组织和癌旁组织,对蛋白组织进行裂解研磨、蛋白电泳、组织酶解、肽段标记、分级标记、高效液相色谱系统分离,采用Q-Exactive质谱仪完成质谱分析。定量分析及生物信息学分析、筛选预后相关蛋白分子。 结果 建立下咽鳞癌蛋白组学数据库:蛋白质7 395个,肽段63 190个,筛选出的差异蛋白上调36个、下调43个,差异表达蛋白质总数79个。完成层次聚类分析基因本体论和京都基因与基因组百科全书信号通路富集分析、蛋白互做分析,筛选出关键蛋白MRPS5,为下咽鳞癌预后的特异性靶分子。 结论 MRPS5可能为下咽鳞状细胞癌的预后靶分子。

关键词: 下咽鳞癌, 蛋白质组学, 非数据依赖性采集, 预后因子

Abstract: Objective To screen specific target molecules related to the prognosis of advanced hypopharyngeal squamous cell carcinoma using non-data dependent acquisition(DIA)proteomics technology. Methods A total of 5 pairs of advanced hypopharyngeal squamous cell carcinoma and adjacent tissues diagnosed in Shanghai General Hospital during Jan. 2016 and Sep. 2018 were selected. The protein tissues were lysed and ground. Protein electrophoresis, tissue enzymatic hydrolysis, peptide labeling, graded labeling, and high-performance liquid chromatography were performed. Mass spectrometry was performed with Q-Exactive mass spectrometer. Quantitative analysis and bioinformatics analysis were conducted to screen prognosis protein molecules. Quantitative analysis and bioinformatics analysis were used to screen prognosis related protein molecules. Results A proteomics database was established, including 7,395 proteins, 63,190 peptide segments, and 79 differentially expressed proteins(36 up-regulated and 43 down-regulated). After GO and KEGG enrichment, PPI protein network interaction and prognosis analysis, the key protein MPRS5 was selected as the specific target molecule for the prognosis of advanced hypopharyngeal squamous cell carcinoma. Conclusion MRPS5 may become a prognostic target molecule for hypopharyngeal squamous cell carcinoma.

Key words: Hypopharyngeal squamous cell carcinoma, Proteomics, Non-data dependent acquisition, Prognostic factors

中图分类号: 

  • R739.63
[1] Bien S, Kaminski B, Zylka S, et al. The evolution of epidemiology and clinical characteristics of laryngeal and hypopharyngeal carcinoma in Poland from 1991 to 2001 [J]. Otolaryngol Pol, 2005, 59(2): 169-181.
[2] Fakhry C, Psyrri A, Chaturvedhi A. HPV and head and neck cancers: state-of-the-science [J]. Oral Oncol, 2014, 50(5): 353-355.
[3] Ferlay J, Soerjomataram I, Dikshit R, et al. Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012 [J]. Int J Cancer, 2015, 136(5): E359-E386.
[4] Simard EP, Torre LA, Jemal A. International trends in head and neck cancer incidence rates: differences by country, sex and anatomic site [J]. Oral Oncol, 2014, 50(5): 387-403.
[5] Byeon HK, Ku M, Yang J. Beyond EGFR inhibition: multilateral combat strategies to stop the progression of head and neck cancer [J]. Exp Mol Med, 2019, 51(1): 1-14.
[6] Gupta S, Kong W, Peng Y, et al. Temporal trends in the incidence and survival of cancers of the upper aerodigestive tract in Ontario and the United States [J]. Int J Cancer, 2009, 125(9): 2159-2165.
[7] 沈枫林, 熊悦婷, 刘晓慧, 等. 基于质谱的蛋白质绝对定量研究策略和建议[J]. 质谱学报, 2021, 42(2): 101-117. SHEN Fenglin, XIONG Yueting, LIU Xiaohui, et al. Strategies and recommendations for the absolute quantification of target proteins based on LC-MS/MS[J]. Journal of Chinese Mass Spectrometry Society, 2021, 42(2): 101-117.
[8] Macmullan MA, Dunn ZS, Graham N, et al. Quantitative proteomics and metabolomics reveal biomarkers of disease as potential immunotherapy targets and indicators of therapeutic efficacy [J]. Theranostics, 2019, 9(25): 7872-7888.
[9] Meyer JG, Schilling B. Clinical applications of quantitative proteomics using targeted and untargeted data-independent acquisition techniques [J]. Expert Rev Proteomics, 2017, 14(5): 419-429.
[10] 陶磊, 周梁, 吴海涛, 等. 154例cN0/cN1下咽鳞状细胞癌颈淋巴清扫效果及预后分析[J]. 中华耳鼻咽喉头颈外科杂志, 2019, 54(4): 272-279. TAO Lei, ZHOU Liang, WU Haitao, et al. Outcome of neck dissection for 154 cN0/cN1 patients with squamous cell carcinoma of hypopharynx and prognostic analysis[J]. Chinese Journal of Otorhinolaryngology Head and Neck Surgery, 2019, 54(4): 272-279.
[11] Wang LT, Liu KJ, Sytwu HK, et al. Advances in mesenchymal stem cell therapy for immune and inflammatory diseases: use of cell-free products and human pluripotent stem cell-derived mesenchymal stem cells [J]. Stem Cells Transl Med, 2021, 10(9): 1288-1303.
[12] Jiang C, Zhang N, Hu X, et al. Tumor-associated exosomes promote lung cancer metastasis through multiple mechanisms [J]. Mol Cancer, 2021, 20(1): 117.
[13] Shui SS, Yao H, Jiang ZD, et al. The differences of muscle proteins between neon flying squid(Ommastrephes bartramii)and jumbo squid(Dosidicus gigas)mantles via physicochemical and proteomic analyses [J]. Food Chem, 2021, 364: 130374. doi: 10.1016/j.foodchem.2021.130374.
[14] Xu W, Anwaier A, Ma C, et al. Multi-omics reveals novel prognostic implication of SRC protein expression in bladder cancer and its correlation with immunotherapy response [J]. Ann Med, 2021, 53(1): 596-610.
[15] Liu J, Zhu W, Li Z, et al. Proteomic analysis of hypop haryngeal and laryngeal squamous cell carcinoma sheds light on differences in survival [J]. Sci Rep, 2020, 10(1): 19459.
[16] Burian A, Lujber L, Gerlinger I, et al. Label-free semiquantitative liquid chromatography-tandem mass spectrometry proteomics analysis of laryngeal/hypopharyngeal squamous cell carcinoma on formalin-fixed, paraffin-embedded tissue samples - a pilot study [J]. Pathol Oncol Res, 2020, 26(4): 2801-2807.
[17] Zhu G, Cai G, Liu Y, et al. Quantitative iTRAQ LC-MS/MS proteomics reveals transcription factor crosstalk and regulatory networks in hypopharyngeal squamous cell carcinoma [J]. J Cancer, 2014, 5(7): 525-536.
[18] Zhou L, Cheng L, Tao L, et al. Detection of hypopharyngeal squamous cell carcinoma using serum proteomics [J]. Acta Otolaryngol, 2006, 126(8): 853-860.
[19] 冉冰冰, 梁楠, 孙辉. 基于质谱的高通量蛋白质组学技术探索肿瘤蛋白标志物的研究进展[J]. 中国肿瘤临床, 2020, 47(8): 411-417. RAN Bingbing,LIANG Nan,SUN Hui. Research progress on tumor protein biomarkers using high throughput proteomics based on mass spectrometry[J]. Chinese Journal of Clinical Oncology, 2020, 47(8): 411-417.
[20] Lee J, Seol M, Jeong S, et al. A Metabolic phenotype based on mitochondrial ribosomal protein expression as a predictor of lymph node metastasis in papillary thyroid carcinoma [J]. Medicine, 2015, 94(2): e380. doi:10.1097/MD.0000000000000380.
[21] Akbergenov R, Duscha S, Fritz AK, et al. Mutant MRPS5 affects mitoribosomal accuracy and confers stress-related behavioral alterations [J]. EMBO reports,2018, 19(11): e46193. doi:10.15252/embr.201846193.
[22] Wei Z, Jia J, Heng G, et al. Sirtuin‐1/Mitochondrial ribosomal protein S5 axis enhances the metabolic flexibility of liver cancer stem cells [J]. Hepatology, 2019, 70(4): 1197-1213.
[23] Houtkooper RH, Mouchiroud L, Ryu D, et al. Mitonuclear protein imbalance as a conserved longevity mechanism [J]. Nature, 2013, 497(7450): 451-457.
[1] 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.
[2] 张华宇,殷思源,刘健,马嘉旭,宋茹,曹国起,王一兵. 氧糖剥夺条件下培养表皮干细胞的定量蛋白质组学分析[J]. 山东大学学报 (医学版), 2021, 59(4): 17-27.
[3] 刘亚飞, 张奇舒, 袁首道, 陈健行. HPV永生化细胞系与SiHa细胞系蛋白质表达差异的质谱分析[J]. 山东大学学报(医学版), 2015, 53(3): 22-26.
[4] 王小岳, 王道光, 郭继生, 李翠玲, 荆瑞瑞, 吕鑫, 陈新骏, 杨静华. 非标记定量法对乙醇诱导肝损伤的蛋白质组学研究[J]. 山东大学学报(医学版), 2015, 53(3): 12-16.
[5] 朱晓丽1,郭淑玲1,苏磊1,冯玉新2,袁方曙1. 蠕形螨全蛋白提取及相对分子量鉴定[J]. 山东大学学报(医学版), 2014, 52(5): 58-62.
[6] 张焕丽1,崔亚洲2,周小艳2,尚伟1. 人类癫痫脑组织的蛋白质组学研究[J]. 山东大学学报(医学版), 2013, 51(3): 80-84.
[7] 王海滨,马伟明,萧畔,马天加,张怀强. 热休克蛋白60在多西紫杉醇治疗激素非依赖性前列腺癌过程中的差异表达[J]. 山东大学学报(医学版), 2012, 50(8): 46-50.
[8] 张华1,3, 刘雪霞2, 宋西成3,雷大鹏1,潘新良1,张庆泉3. T1期声门区喉癌差异蛋白质组学的初步分析[J]. 山东大学学报(医学版), 2012, 50(2): 110-115.
[9] 萧畔1,张怀强1,崔亚洲2,赵升田1,马天加1,周春文1. 多西紫杉醇诱导前列腺癌PC3细胞凋亡的蛋白质组学研究[J]. 山东大学学报(医学版), 2012, 50(10): 77-.
[10] . 前列腺癌细胞LNCaP中雄激素依赖性和雄激素非依赖性蛋白差异性表达研究[J]. 山东大学学报(医学版), 2009, 47(9): 62-65.
[11] . 乳腺癌转移相关蛋白蛋白质组学初步研究[J]. 山东大学学报(医学版), 2009, 47(7): 61-64.
[12] 柏淑美,刘师莲,杨银荣,郭绪晓,秦延江,邓小梅 . RRMS疾病潜在生物标记物关系探讨[J]. 山东大学学报(医学版), 2008, 46(4): 340-343.
[13] 秦延江,刘师莲,杨银荣,柏淑美,张旭华,邓小梅 . 中枢神经系统脱髓鞘疾病脑脊液蛋白质组学研究[J]. 山东大学学报(医学版), 2008, 46(1): 9-14.
[14] 靖永胜,刘师莲,张旭华,吴晓青,杨银荣,王伟 . 肾癌及癌旁组织蛋白质组学的比较研究[J]. 山东大学学报(医学版), 2007, 45(8): 781-784.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
[1] 王焕亮,孙宝柱,杜洪玫,周长青,张丽. 不同麻醉监测指标调控异丙酚麻醉的比较[J]. 山东大学学报(医学版), 2006, 44(5): 471 -474 .
[2] 宋永红,马春红,吕红娟,朱传福,聂向民,王玫,刘艳,张萍 . 中国北方汉族人群HLA基因多态性研究[J]. 山东大学学报(医学版), 2007, 45(6): 546 -553 .
[3] 黄庆,田辉,李林,梁飞,刘贤锡 . 老年人肺癌组织中鸟氨酸脱羧酶基因表达及其临床意义[J]. 山东大学学报(医学版), 2006, 44(6): 556 -559 .
[4] 朱梅佳,韩巨,王新怡,鹿伟,王爱华,关心华,曹霞,曹秉振. 伴有皮层下梗死和白质脑病的常染色体显性遗传性脑动脉病临床病理研究[J]. 山东大学学报(医学版), 2006, 44(8): 834 -839 .
[5] 宋海岩,武玉玲,张艳萍. 牡蛎提取物对高温致神经管畸形中凋亡细胞的保护作用[J]. 山东大学学报(医学版), 2007, 45(2): 113 -116 .
[6] 于慧1,2 ,陈少华1 ,赵家军2 ,高聆3
. 乙醇对人肝L02细胞糖原和GSK3β、PAMPK的影响[J]. 山东大学学报(医学版), 2009, 47(04): 75 -78 .
[7] 侯晓阳,卜培莉,张运,冯进波,刘春喜,李传保,郝明秀. 过氧化酶体增殖物激活受体α抑制血管紧张素Ⅱ促心肌纤维化作用的试验研究[J]. 山东大学学报(医学版), 2007, 45(7): 665 -668 .
[8] 曾季平,王丽娜,王立祥,任晓辉,张孟业,夏文,崔行. 氯化锰致PC12细胞损伤的研究[J]. 山东大学学报(医学版), 2006, 44(5): 467 -470 .
[9] 舒雅,齐峰. 下颌角托在肥胖患者全麻诱导中的应用[J]. 山东大学学报(医学版), 2007, 45(10): 1072 -1074 .
[10] 王志刚,丁 璇,孙 鹏/sup>,王成伟,郝晓光,潘 顺 . 术前脑血管造影在血管内支架成形术治疗缺血性脑血管病中的应用[J]. 山东大学学报(医学版), 2007, 45(2): 146 -148 .