下咽癌中差异表达的蛋白激酶及其抑制剂的生物信息学筛选

doi:10.6040/j.issn.1673-3770.0.2015.340

摘要/Abstract

摘要： 目的筛选下咽癌中差异表达的激酶基因及其选择性抑制剂,为下咽癌的分子靶向治疗提供新的参考。方法利用GEO数据库和SAM软件筛选下咽癌中差异表达的激酶基因,体外培养人下咽癌FaDu细胞系。为验证GEO数据库中芯片结果的准确性,利用实时定量聚合酶链反应(Real-time PCR)检测差异表达激酶在FaDu细胞中的表达量,通过KEGG数据库获得激酶调控的通路,利用激酶抑制剂数据库和文献挖掘筛选获得在下咽癌FaDu细胞系中差异表达激酶的选择性抑制剂。结果 (1)在GEO数据库的下咽癌基因组表达谱中,共筛选出3个高表达的激酶基因,分别为PKC-β、CDK6和CDC42(差异倍数≥2.0且P<0.05);(2)Real-time PCR结果显示在人下咽癌FaDu细胞中这3个上调激酶基因也出现高表达(P<0.05),证明全基因组的结果准确;(3)KEGG通路分析的结果显示3个高表达激酶调控复杂的通路网络;(4)激酶抑制剂的筛选结果显示共有5个激酶抑制剂调控PKC-β, 4个激酶抑制剂调控CDK6,3个激酶抑制剂调控CDC42。文献挖掘的结果显示在这12个激酶抑制剂中,有4个在癌症方面的研究较少,文献<10篇。结论下咽癌中共有3个激酶PKC-β、CDK6和CDC42发生高表达,并发挥促癌作用。它们的激酶抑制剂可能有潜在的抗癌作用,为下咽癌的分子治疗提供新的切入点。

关键词: 下咽癌, 蛋白激酶, 激酶抑制剂, 生物信息学

Abstract: Objective To screen differentially expressed protein kinases and their inhibitors, in order to provide new targets for molecular therapy of hypopharyngeal cancer. Methods GEO database and SAM software were employed to screen the differentially expressed protein kinase genes in hypopharyngeal cancer. Human hypopharyngeal cancer FaDu cell line was cultured in vitro. Real-time PCR was used to prove the accuracy of microarray results. Based on KEGG database, the kinase-regulated pathways were identified. The inhibitors of such kinases were identified by using kinase-inhibitor database. Results A total of 3 protein kinase genes(PKC-β, CDK6 and CDC42)were identified(fold change≥2.0, P<0.05). Real-time PCR showed that the 3 kinase genes were differentially expressed in FaDu cells, which was consistent with the microarray results(P<0.05)Pathway analysis indicated that a complex pathway network was regulated by the 3 kinases. The results of inhibitor screening showed that 5 inhibitors regulated PKC-β, 4 inhibitors regulated CDK6 and 3 inhibitors regulated CDC42. There were less than 10 studies about the 4 inhibitors in cancer. Conclusion Three differentially expressed protein kinases(PKC-β, CDK6 and CDC42)are identified and they are able to promote the development of tumor. Their kinase inhibitors may play a potential anti-cancer role, which may provide a new target for molecular therapy of hypopharyngeal cancer.

Key words: Bioinformatics, Protein kinase, Kinase inhibitor, Hypopharyngeal cancer

中图分类号:

R739.65

胡文良,郑艳秋,崔晓波,崔彦茹,孙源昊. 下咽癌中差异表达的蛋白激酶及其抑制剂的生物信息学筛选[J]. 山东大学耳鼻喉眼学报, 2016, 30(3): 24-28.

HU Wenliang, ZHENG Yanqiu, CUI Xiaobo, CUI Yanru, SUN Yuanhao. Bioinformatic screening of differentially expressed protein kinases and their inhibitors in hypopharyngeal cancer.[J]. JOURNAL OF SHANDONG UNIVERSITY (OTOLARYNGOLOGY AND OPHTHALMOLOGY), 2016, 30(3): 24-28.

参考文献

[1] Gourin C G, Terris D J. Carcinoma of the hypopharynx[J]. Surg Oncol Clin N Am, 2004, 13(1):81-98.
[2] Buckley J G, MacLennan K. Cervical node metastases in laryngeal and hypopharyngeal cancer: a prospective analysis of prevalence and distribution[J]. Head Neck, 2000, 22(4):380-385.
[3] American Society of Clinical Oncology, Pfister D G, Laurie S A, et al. American Society of Clinical Oncology clinical practice guideline for the use of larynx-preservation strategies in the treatment of laryngeal cancer[J]. J Clin Oncol, 2006, 24(22):3693-3704.
[4] Piirsoo A, Kasak L, Kauts M L, et al. Protein kinase inhibitorSU6668 attenuates positive regulation of Gli proteins in cancer[J]. Biochim Biophys Acta, 2014, 1843(4):703-714.
[5] Manning G, Whyte D B, Martine R, et al. The protein kinase complement of the human genome[J]. Science, 2002, 298(5600):1912-1934.
[6] Okkenhaug K. Signaling by the phosphoinositide 3-kinase family inimmune[J]. Annu Rev Immunol, 2013, 31:675-704.
[7] Kizub I V, Klymenko K I, Soloviev A I. Protein kinase C in enhanced vascular tone in diabetes mellitus[J]. Int J Cardiol, 2014, 174(2):230-242.
[8] Barrett T, Suzek T O, Troup D B, et al. NCBI GEO: mining millions of expression profiles-database and tools[J]. Nucleic Acids Res, 2005, 33(Database issue):D562-566.
[9] Joost S, Kalbermatten M, Bonin A. Spatial analysis method(sam): a software tool combining molecular and environmental data to identify candidate loci for selection[J]. Mol Ecol Resour, 2008, 8(5):957-960.
[10] Basson M D, Zeng B, Downey C, et al. Increased extracellular pressure stimulates tumor proliferation by a mechanosensitive calcium channel and PKC-β[J]. MolOncol, 2015, 9(2):513-526.
[11] He L F, Wang T T, Gao Q Y, et al. Stanniocalcin-1 promotes tumor angiogenesis through up-regulation of VEGF in gastric cancer cells[J]. J Biomed Sci, 2011, 18(1):39-48.
[12] Fridberg M, Kjellström S, Anagnostaki L, et al. Immunohistochemical analyses of phosphatases in childhood B-cell lymphoma: lower expression of PTEN and HePTP and higher number of positive cells for nuclear SHP2 in B-cell lymphoma cases compared to controls[J]. Pediatr Hematol Oncol, 2008, 25(6):528-540.
[13] Xiao F, Zhang W, Zhou L, et al. microRNA-200a is an independent prognostic factor of hepatocellular carcinoma and induces cell cycle arrest by targeting CDK6[J]. Oncol Rep, 2013, 30(5):2203-2210.
[14] Xia B, Yang S, Liu T, et al. miR-211 suppresses epithelial ovarian cancer proliferation and cell-cycle progression by targeting Cyclin D1 and CDK6[J]. Mol Cancer, 2015, 14(1):57-70.
[15] Comstock C E, Augello M A, Goodwin J F, et al. Targeting cell cycle and hormone receptor pathways in cancer[J]. Oncogene, 2013, 32(48):5481-5491.
[16] Zen Y, Harada K, Sasaki M, et al. Intrahepatic cholangio carcinoma escapes from growth inhibitory effect of transforming growth factor-beta1 by overexpression of cyclin D1[J]. Lab Invest, 2005, 85(4):572-581.
[17] Wang J, Li X, Lu X, et al. The regulation of stat3 signal transduction pathway to G1 to S phase of laryngo carcinoma cell[J]. Clin J Otorhinolarynol Head Neck Surg, 2008, 22(15):699-703.
[18] Kwon D Y, Lee H E, Weitzel D H, et al. Synthesis and Biological Evaluation of Manassantin Analogues for Hypoxia-Inducible Factor 1α Inhibition[J]. J Med Chem, 2015, 58(19):7659-7671.
[19] Zupa A, Improta G, Silvestri A, et al. A pilot characterization of human lung NSCLC by protein pathway activation mapping[J]. J Thorac Oncol, 2012, 7(12):1755-1766.
[20] Razidlo G L, Magnine C, Sletten A C, et al. Targeting Pancreatic Cancer Metastasis by Inhibition of Vav1, a Driver of Tumor Cell Invasion[J]. Cancer Res, 2015, 75(14):2907-2915.
[21] Cheung C H, Coumar M S, Chang J Y, et al. Aurora kinase inhibitor patents and agents in clinical testing: an update(2011-13)[J]. Expert Opin Ther Pat, 2014, 24(9):1021-1038.
[22] Murphy C G, Dickler M N. The Role of CDK4/6 Inhibition in Breast Cancer[J]. Oncologist, 2015, 20(5):483-490.
[23] Chen C, Song X, Ma S, et al. Cdc42 inhibitor ML141 enhances G-CSF-induced hematopoietic stem and progenitor cell mobilization[J]. Int J Hematol, 2015, 101(1):5-12.
[24] Gray J E, Altiok S, Alexandrow M G, et al. Phase 2 randomized study of enzastaurin(LY317615)for lung cancer prevention in former smokers[J]. Cancer, 2013, 119(5):1023-1032.
[25] Rampling R, Sanson M, Gorlia T, et al. A phase I study of LY317615(enzastaurin)and temozolomide in patients with gliomas(EORTC trial 26054)[J]. Neuro Oncol, 2012, 14(3):344-350.
[26] Blevins D P, Dadu R, Hu M, et al. Aerodigestive fistula formation as a rare side effect of antiangiogenic tyrosine kinase inhibitortherapy for thyroid cancer[J]. Thyroid, 2014, 24(5):918-922.
[27] 潘新良, 雷大鹏, 许风雷, 等. 下咽癌的外科治疗[J]. 山东大学耳鼻喉眼学报, 2007, 21(1):1-6. PAN Xinliang, LEI Dapeng, XU Fenglei, et al. Surgical treatment of lower pharyngeal carcinoma[J]. J Otolaryngol Ophthalmol Shandong Univ, 2007, 21(1):1-6.

相关文章 15

[1]	董频,英信江,陈歆维,邓志宏,张少强,於子卫,金斌,孙臻峰,谢晋,祝江才. 新辅助化疗方案尼妥珠单抗联合奈达铂和5-氟尿嘧啶治疗下咽鳞癌初步临床分析[J]. 山东大学耳鼻喉眼学报, 2016, 30(3): 10-14.
[2]	李晓明,宋琦,李红霞,陶振峰,申宇鹏,肖淑芬. 胸大肌肌皮瓣在侵犯下咽晚期头颈鳞癌手术切除术后缺损修复中的应用[J]. 山东大学耳鼻喉眼学报, 2016, 30(3): 4-9.
[3]	李文明,魏东敏,孙睿杰,钱晔,刘大昱,解光,雷大鹏,潘新良. 下咽癌侵犯颈段食管的治疗[J]. 山东大学耳鼻喉眼学报, 2016, 30(3): 15-19.
[4]	高振峰,陶谦,蔡晓岚. ERK5蛋白在鼻息肉细胞中的表达和意义[J]. 山东大学耳鼻喉眼学报, 2016, 30(1): 40-42.
[5]	江昌,许承弼,辛丁,鞠洋,李鸿燕,王金月. 下咽癌腋窝淋巴结转移1例及文献回顾[J]. 山东大学耳鼻喉眼学报, 2016, 30(1): 90-92.
[6]	杜祥阁, 张营春, 颜昕, 王翠, 赵博军. 下调血管内皮细胞蛋白激酶CK2表达对血管增殖影响的体外研究[J]. 山东大学耳鼻喉眼学报, 2015, 29(3): 76-80.
[7]	郑立友. 中西医结合治疗喉癌下咽癌手术并发咽瘘10例[J]. 山东大学耳鼻喉眼学报, 2012, 26(6): 31-33.
[8]	沈斌，董频，英信江，高尚，李大伟. ABCG2及NF-κB p65在下咽癌中的表达意义[J]. 山东大学耳鼻喉眼学报, 2012, 26(4): 17-20.
[9]	曹汉海1，丁海霞2，彭洪娟3. 多层螺旋CT后处理技术对下咽癌评估的价值[J]. 山东大学耳鼻喉眼学报, 2011, 25(3): 25-27.
[10]	曾志梅1，梁永辉2，杨嶷芳2，窦晓清2，罗碧强2，张丽卫2，陈贤明2. 巨噬细胞移动抑制因子在下咽癌中的表达[J]. 山东大学耳鼻喉眼学报, 2011, 25(1): 17-19.
[11]	叶婷1，马超2，潘新良1. 下咽癌瘤内淋巴管生成及颈淋巴转移关系的研究[J]. 山东大学耳鼻喉眼学报, 2011, 25(1): 24-.
[12]	梁辉1,2，潘新良1,马超3，王启荣2，韩飞2，崔朝阳2. 下咽癌FADU细胞系中CD133阳性细胞侵的袭能力[J]. 山东大学耳鼻喉眼学报, 2010, 24(2): 5-7.
[13]	高英恺1，张凤英1，王树勇1，刘清明2，陈淑萍1. 喉癌及下咽癌手术后咽瘘的治疗及预防[J]. 山东大学耳鼻喉眼学报, 2009, 23(6): 21-23.
[14]	高兴强1 ，赵德安1 ，方正2 ，于修义2 . 非开胸胃代食管术治疗同时性下咽并胸段食管重复癌5例[J]. 山东大学耳鼻喉眼学报, 2009, 23(2): 24-26 .
[15]	潘新良,雷大鹏,许风雷,张立强,栾信庸 . 下咽癌的外科治疗[J]. 山东大学耳鼻喉眼学报, 2007, 21(1): 1-07 .

多维度评价

Viewed

Full text

Abstract

Cited

Shared

Discussed