PDF下载 ( 2591 KB)
基于生物信息学分析核仁纺锤体相关蛋白1在肝细胞癌中的表达及对临床预后的影响
DOI: 10.3969/j.issn.1001-5256.2021.07.024
利益冲突声明：本研究不存在研究者、伦理委员会成员、受试者监护人以及与公开研究成果有关的利益冲突。
作者贡献声明：靳鑫旻、杨通旺、徐庆国负责课题设计，资料分析，撰写论文；靳鑫旻、刘欢、关鸽参与收集数据，修改论文；臧运金等负责拟定写作思路，指导撰写文章并最后定稿。
Expression of nucleolar spindle-associated protein 1 in hepatocellular carcinoma and its effect on clinical prognosis: A bioinformatics analysis
-
摘要:
目的 探讨在肝癌中核仁纺锤体相关蛋白1(NUSAP1)的表达及其对临床预后的影响。 方法 从基因表达数据库(GEO)下载HCC芯片数据集GSE57957、GSE14520、GSE22058、GSE46444、GSE54236、GSE36376、GSE64041、GSE76297、GSE76427、GSE102079和癌症基因组图谱(TCGA)中的HCC RNA-seq数据，利用R 4.0软件分析NUSAP1在肝癌及癌旁组织的表达差异。应用t-分布邻域嵌入算法(tSNE)对单细胞测序数据的降维，分析NUSAP1在肝癌和癌旁组织中的差异表达。收集2018年1月—2019年11月在青岛大学附属医院进行同种异体肝移植术的42例乙型肝炎相关肝癌患者的癌组织及癌旁组织，收集其临床病理资料并分析与NUSAP1基因表达的相关性。计数资料2组间比较采用χ2检验。以Kaplan-Meier方法生存分析NUSAP1表达水平与总体生存率及无病间隔生存率之间的关系，运用log-rank进行检验，通过Graphpad prim 7使其结果可视化。 结果 NUSAP1在GEO数据库与TCGA数据库数据集中的肝癌组织中均高表达(P值均＜0.001)。来源于肿瘤组织的细胞其NUSAP1高表达细胞明显高于癌旁组织细胞。NUSAP1在29例患者的肝癌组织中高表达，在13例患者的肝癌组织中低表达，NUSAP1在肝癌组织中高表达组及低表达组间，Child-Pugh分级、TNM分期、Okuda分期比较差异均有统计学意义(χ2值分别为5.469、6.836、4.617，P值均＜0.05)。NUSAP1高表达的肝癌患者总体生存率与无病间隔生存率显著低于低表达的肝癌患者，NUSAP1低表达患者1、3、5生存率分别为92.96%、83.80%、76.76%，明显高于高表达的患者(76.76%、67.60%、64.78%)，NUSAP1高表达的患者中位生存时间明显低于低表达的患者(61.7个月 vs 108.6个月)(P值均＜0.05)。 结论 NUSAP1在肝癌细胞中高表达，可作为乙型肝炎肝癌患者不良预后的潜在标志物。 -
关键词:
- 癌，肝细胞 /
- 核仁纺锤体相关蛋白1 /
- 计算生物学
Abstract:Objective To investigate the expression of the nucleolar spindle-associated protein 1 (NUSAP1) gene in hepatocellular carcinoma (HCC) and its effect on clinical prognosis. Methods HCC microarray datasets GSE57957, GSE14520, GSE22058, GSE46444, GSE54236, GSE36376, GSE64041, GSE76297, GSE76427, and GSE102079 were downloaded from the Gene Expression Omnibus (GEO) database, and HCC RNA-Seq data were downloaded from The Cancer Genome Atlas (TCGA). R software 4.0 was used to investigate the difference in the expression of NUSAP1 between HCC tissue and adjacent tissue. The t-distributed stochastic neighbour embedding (t-SNE) method was used to perform dimensionality reduction of single-cell sequencing data and analyze the difference in the expression of NUSAP1 between HCC tissue and adjacent tissue. HCC tissue samples and adjacent tissue samples were collected from 42 patients with hepatitis B-related HCC who underwent liver transplantation in The Affiliated Hospital of Qingdao University from January 2018 to November 2019, and related clinical and pathological data were collected to analyze their correlation with the expression of the NUSAP1 gene. The chi-square test was used for comparison of paired data. The Kaplan-Meier method was used to analyze the correlation of the expression level of NUSAP1 with overall survival rate and disease-free survival, and the log-rank test was used for survival difference analysis; the results were visualized using Graphpad prim 7. Results NUSAP1 was highly expressed in HCC tissue in GEO and TCGA datasets (P < 0.001). The cells from HCC tissue had a significantly higher proportion of cells with high NUSAP1 expression than those from adjacent tissue. NUSAP1 showed high expression in HCC tissue of 29 patients and low expression in HCC tissue of 13 patients, and there were significant differences in Child-Pugh class, TNM stage, and Okuda stage between the high-expression group and the low-expression group (χ2=5.469, 6.836 and 4.617, all P < 0.05). The HCC patients with high expression of NUSAP1 had significantly lower overall survival rate and disease-free survival rate than those with low expression; compared with the patients with high expression, the patients with low expression had significantly higher 1-, 3-, and 5-year survival rates (92.96%/83.80%/76.76% vs 76.76%/67.60%/64.78%, P < 0.05), and the patients with high expression had a significantly shorter median survival time than those with low expression (61.7 months vs 108.6 months, P < 0.05). Conclusion NUSAP1 is highly expressed in cells from HCC tissue and may be used as a potential marker for poor prognosis of patients with hepatitis B-related HCC. -
表 1 NUSAP1表达水平与肝癌患者临床病理特征的关系
项目 高表达(n=29) 低表达(n=13) χ2值 P值 性别(例) 0.022 0.88 男 24 11 女 5 2 血型(例) 0.628 0.89 O型 5 3 A型 11 5 B型 12 5 AB型 1 0 年龄 2.143 0.14 ＜50岁 16 4 ≥50岁 13 9 BMI 0.633 0.43 18.5~23.9 kg/m2 14 8 ＜18.5 kg/m2或≥24 kg/m2 15 5 AFP(例) 0.252 0.62 ＜20 ng/ml 11 6 ≥20 ng/ml 18 7 MELD评分(例) 0.032 0.86 ＜20分 17 8 ≥30分 12 5 Child-Pugh分级(例) 5.469 0.02 A级 7 8 B或C级 22 5 肿瘤数目(例) 0.001 0.97 单发 11 5 多发 18 8 TNM分期(例) 6.836 0.01 Ⅰ期 3 6 Ⅱ或Ⅲ期 26 7 Okuda分期(例) 4.617 0.03 Ⅰ 6 7 Ⅱ或Ⅲ 23 6 
下载: 导出CSV
-
[1] BRAY F, FERLAY J, SOERJOMATARAM I, et al. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA Cancer J Clin, 2018, 68(6): 394-424. DOI: 10.3322/caac.21492. [2] KULIK L, EL-SERAG HB. Epidemiology and management of hepatocellular carcinoma[J]. Gastroenterology, 2019, 156(2): 477-491. e1. DOI: 10.1053/j.gastro.2018.08.065. [3] WANG SY, FENG LY, MENG ZQ. Bicluster and pathway enrichment analysis related to tumor progression of hepatocellular carcinoma[J]. Eur Rev Med Pharmacol Sci, 2015, 19(7): 1191-1197. http://europepmc.org/articles/PMC4237098 [4] BAATARKHUU O, GERELCHIMEG T, MUNKH-ORSHIKH D, et al. Epidemiology, genotype distribution, prognosis, control, and management of viral hepatitis B, C, D, and hepatocellular carcinoma in mongolia[J]. Euroasian J Hepatogastroenterol, 2018, 8(1): 57-62. DOI: 10.5005/jp-journals-10018-1260. [5] EL-SERAG HB. Epidemiology of viral hepatitis and hepatocellular carcinoma[J]. Gastroenterology, 2012, 142(6): 1264-1273. e1. DOI: 10.1053/j.gastro.2011.12.061. [6] FENG WG, WU W, GONG JP. Research progress on therapy of hepatocellular carcinoma[J]. Chin J Curr Adv Gen Surg, 2013, 16(7): 583-585. DOI: 10.3969/j.issn.1009-9905.2013.07.026.冯文贵, 吴伟, 龚建平. 肝细胞癌治疗的研究进展[J]. 中国现代普通外科进展, 2013, 16(7): 583-585. DOI: 10.3969/j.issn.1009-9905.2013.07.026. [7] RAEMAEKERS T, RIBBECK K, BEAUDOUIN J, et al. NuSAP, a novel microtubule-associated protein involved in mitotic spindle organization[J]. J Cell Biol, 2003, 162(6): 1017-1029. DOI: 10.1083/jcb.200302129. [8] PETRY S. Mechanisms of mitotic spindle assembly[J]. Annu Rev Biochem, 2016, 85: 659-683. DOI: 10.1146/annurev-biochem-060815-014528. [9] ZHOU Q, LEE KJ, KURASAWA Y, et al. Faithful chromosome segregation in Trypanosoma brucei requires a cohort of divergent spindle-associated proteins with distinct functions[J]. Nucleic Acids Res, 2018, 46(16): 8216-8231. DOI: 10.1093/nar/gky557. [10] LIU Z, GUAN C, LU C, et al. High NUSAP1 expression predicts poor prognosis in colon cancer[J]. Pathol Res Pract, 2018, 214(7): 968-973. DOI: 10.1016/j.prp.2018.05.017. [11] CHEN B, LI Y, WEI YH, et al. Expression and clinical significance of NUSAP1 in non-small cell lung cancer[J]. Int J Respir, 2017, 37(22): 1681-1684. DOI: 10.3760/cma.j.issn.1673-436X.2017.22.001.陈波, 李悦, 位云艳, 等. 非小细胞肺癌中NUSAP1的表达及临床意义[J]. 国际呼吸杂志, 2017, 37(22): 1681-1684. DOI: 10.3760/cma.j.issn.1673-436X.2017.22.001. [12] GAO S, YIN H, TONG H, et al. Nucleolar and spindle associated protein 1 (NUSAP1) promotes bladder cancer progression through the TGF-β signaling pathway[J]. Onco Targets Ther, 2020, 13: 813-825. DOI: 10.2147/OTT.S237127. [13] GE Y, LI Q, LIN L, et al. Downregulation of NUSAP1 suppresses cell proliferation, migration, and invasion via inhibiting mTORC1 signalling pathway in gastric cancer[J]. Cell Biochem Funct, 2020, 38(1): 28-37. DOI: 10.1002/cbf.3444. [14] GORDON CA, GULZAR ZG, BROOKS JD. NUSAP1 expression is upregulated by loss of RB1 in prostate cancer cells[J]. Prostate, 2015, 75(5): 517-526. DOI: 10.1002/pros.22938. [15] ZHANG X, PAN Y, FU H, et al. Nucleolar and spindle associated protein 1 (NUSAP1) inhibits cell proliferation and enhances susceptibility to epirubicin in invasive breast cancer cells by regulating cyclin D kinase (CDK1) and DLGAP5 expression[J]. Med Sci Monit, 2018, 24: 8553-8564. DOI: 10.12659/MSM.910364. [16] XIE Q, OU-YANG W, ZHANG M, et al. Decreased expression of NUSAP1 predicts poor overall survival in cervical cancer[J]. J Cancer, 2020, 11(10): 2852-2863. DOI: 10.7150/jca.34640. [17] LI Y, HUANG C, DING L, et al. Deep learning in bioinformatics: Introduction, application, and perspective in the big data era[J]. Methods, 2019, 166: 4-21. DOI: 10.1016/j.ymeth.2019.04.008. [18] SONG B, TANG JW, WANG B, et al. Identify lymphatic metastasis-associated genes in mouse hepatocarcinoma cell lines using gene chip[J]. World J Gastroenterol, 2005, 11(10): 1463-1472. DOI: 10.3748/wjg.v11.i10.1463. [19] ZHOU YL, LIU JY, YAO HW, et al. Increased expression of lncFOXC1 promotes liver cancer formation[J]. Chin J Clin Pharmacol Ther, 2019, 24(1): 27-31. DOI: 10.12092/j.issn.1009-2501.2019.01.005.周玉玲, 刘激扬, 姚红伟, 等. lncFOXC1表达增加促进肝癌发生[J]. 中国临床药理学与治疗学, 2019, 24(1): 27-31. DOI: 10.12092/j.issn.1009-2501.2019.01.005. [20] YUAN SX, ZHOU WP. Progress and hot spots of comprehensive treatment for primary liver cancer[J]. Chin J Dig Surg, 2021, 20(2): 163-170. DOI: 10.3760/cma.j.cn115610-20201211-00776.袁声贤, 周伟平. 原发性肝癌综合治疗的进展和热点[J]. 中华消化外科杂志, 2021, 20(2): 163-170. DOI: 10.3760/cma.j.cn115610-20201211-00776. 期刊类型引用(16)
1. 巴明玉，潘研，王娴，陈亚琳，燕树勋. 强肝胶囊对非酒精性脂肪肝肥胖患者糖脂代谢水平及肝功能的影响. 中医学报. 2024(02): 416-420 . 
百度学术2. 范文，郑垂仰，周旋，孔怡琳，潘丰满，杨钦河，张玉佩. 6种中成药治疗非酒精性脂肪性肝病有效性和安全性的网状Meta分析. 中药新药与临床药理. 2023(11): 1612-1622 . 百度学术3. 崔姗姗，刘英，江霞. 非酒精性脂肪性肝病与胰岛素抵抗的研究进展. 医学信息. 2022(10): 66-69 . 百度学术4. 舒祥兵，操颖，顾晔，杨志新，季光，张莉. 强肝胶囊促进肝糖原合成降低大鼠高血糖的机制研究. 上海中医药大学学报. 2021(03): 45-51 . 百度学术5. 贾英杰，张晓博，张新元，杨红霞，郭争荣. 强肝胶囊对猪血清诱导大鼠肝纤维化的治疗作用. 河北医药. 2021(20): 3104-3107 . 百度学术6. 朱丹，孙婷婷，陈兰羽，谢铮. 脂肪性肝病中医药认识. 辽宁中医药大学学报. 2020(05): 70-73 . 百度学术7. 崔翔，刘玲兰，华鹏，郭薇，薛敬东. 从脾论治非酒精性脂肪性肝病的体会与研究概况. 中医临床研究. 2020(14): 136-138 . 百度学术8. 郭明升，刘娟，郭英杰，籍芳华，郝艳爽. 阿托伐他汀联用脂康颗粒或强肝胶囊用于脑梗死二级预防效果比较. 现代中西医结合杂志. 2020(22): 2400-2404+2446 . 百度学术9. 王秀琴. 强肝胶囊联合甘草酸二铵肠溶胶囊治疗非酒精性脂肪肝的效果. 河南医学研究. 2019(06): 1089-1091 . 百度学术10. 黄静，陈湘清，郭东升，吴奋. 同步抗感染治疗对合并幽门螺杆菌感染非酒精性脂肪肝病患者治疗结局的影响研究. 重庆医学. 2018(01): 97-100 . 百度学术11. 陈少颖，易新宇，周小博，张荣臻. 中医药治疗非酒精性脂肪肝的研究进展. 大众科技. 2018(06): 68-70 . 百度学术12. 周桃梅，杨上文，汪剑波. 强肝胶囊联合二甲双胍治疗非酒精性脂肪肝的疗效及其对Nod样受体-3炎症小体的影响. 中国医院统计. 2018(03): 206-208 . 百度学术13. 柳琳琳，毛德文，吕建林，黄瑞. 强肝胶囊对非酒精性脂肪性肝病患者疗效及安全性的Meta分析. 中成药. 2018(08): 1715-1720 . 百度学术14. 李滨，张佳圆，王凤永，李春晖，齐丽韫，王玉华，赵培利. 胆宁片治疗非酒精性脂肪肝临床效果及对血脂、肝功能和肝纤维化的影响. 解放军医药杂志. 2018(10): 93-96 . 百度学术15. 朱紫馨，祝丽丽，程明亮. 同步抗感染治疗非酒精性脂肪肝合并幽门螺杆菌感染的效果观察. 临床合理用药杂志. 2018(31): 58-59 . 百度学术16. 李军祥，王允亮，郭一，谢添弘. 中医药治疗非酒精性脂肪性肝病专题笔谈. 中国中西医结合消化杂志. 2017(11): 801-804 . 百度学术其他类型引用(13)
-
百度学术
