乳腺癌MDA-MB-231细胞中Rab7a基因的网络分析

doi:10.13481/j.1671-587x.20200306

吉林大学学报(医学版) ›› 2020, Vol. 46 ›› Issue (03): 464-469.doi: 10.13481/j.1671-587x.20200306

• 基础研究 • 上一篇

乳腺癌MDA-MB-231细胞中Rab7a基因的网络分析

徐凤英¹, 刘儒¹, 马丽¹, 曲一帆¹, 肖伟利², 王玉珍³, 谢基明²

1. 内蒙古医科大学研究生学院, 内蒙古呼和浩特 010110;
2. 内蒙古自治区人民医院检验科, 内蒙古呼和浩特 010010;
3. 内蒙古农业大学生命科学学院, 内蒙古呼和浩特 010018

收稿日期:2019-06-04 发布日期:2020-06-11
通讯作者: 谢基明,主任医师,硕士研究生导师(Tel:0471-3283999,E-mail:xclinton@sina.com) E-mail:xclinton@sina.com
作者简介:徐凤英(1994-),女,内蒙古自治区呼和浩特市人,在读医学硕士,主要从事炎症的免疫与分子机制方面的研究。
基金资助:
国家自然科学基金资助课题（81360394，31270922）

Network analysis on Rab7a gene in breast cancer MDA-MB-231 cells

XU Fengying¹, LIU Ru¹, MA Li¹, QU Yifan¹, XIAO Weili², WANG Yuzhen³, XIE Jiming²

1. Graduate School, Inner Mongolia Medical University, Hohhot 010110, China;
2. Department of Clinical Laboratory, People's Hospital, Inner Mongolia Autonomons Region, Hohhot 010010, China;
3. College of Life Science, Inner Mongolia Agricultural University, Hohhot 010018, China

Received:2019-06-04 Published:2020-06-11

摘要/Abstract

摘要： 目的：探讨三阴性乳腺癌（TNBC） MDA-MB-231细胞中Rab7a基因敲除后相关基因表达的变化，阐明Rab7a相关基因在TNBC中的作用。方法：选取对数生长期的乳腺癌ZR-75-30、MCF-7、T-47D、MDA-MB-231和HCC-1937细胞，采用Western blotting法检测乳腺癌ZR-75-30、MCF-7、T-47D、MDA-MB-231和HCC-1937细胞中Rab7a蛋白表达水平。慢病毒敲除TNBCMDA-MB-231细胞中Rab7a基因，分为阴性对照组和Rab7a基因敲除组，根据敲除的4种不同Rab7a序列分为KD1组、KD2组、KD3组和KD4组。采用qPCR法检测各组MDA-MB-231细胞中Rab7a基因敲除效率，全基因表达谱芯片检测Rab7a基因敲除效率最高组（KD2组）和阴性对照组差异基因，一体化通路分析（IPA）软件分析Rab7a基因与其他基因的相互作用。结果：在TNBCMDA-MB-231细胞中可见Rab7a蛋白表达。与阴性对照组比较，KD2组乳腺癌MDA-MB-231细胞中Rab7a基因敲除效率最高（P<0.01）；与阴性对照组比较，KD2组乳腺癌MDA-MB-231细胞中有634个差异基因，其中上调基因和下调基因数均增多（P<0.01）；差异基因分析，Rab7a基因敲除与癌症、细胞存活、细胞周期和细胞生长及转移等有密切联系；Rab7a的IPA网络图中eIF4F、IRS1和RPS6KB1表达下调，PRKAA1、IKBKE、VEGFA和转录因子ATF2表达上调。结论：Rab7a基因在TNBCMDA-MB-231细胞中以基因网络的形式发挥作用；Rab7a基因与多基因相互作用，参与癌细胞的病理过程。

关键词: 乳腺肿瘤, Rab7a, 三阴性乳腺癌, MDA-MB-231细胞, 一体化通路分析

Abstract: Objective: To investigate the changes of expressions of the related genes in the triple-negative breast cancer(TNBC) MDA-MB-231 cells after knockout of Rab7a gene, and to elucidate the roles of Rab7a-related genes in TNBC. Methods: The breast cancer ZR-75-30, MCF-7, T-47D, MDA-MB-231 and HCC-1937 cells in the logarithmic phase were selected.The expression levels of Rab7a protein in the breast cancer ZR-75-30, MCF-7, T-47D, MDA-MB-231, and HCC-1937 cells were detected by Western blotting method. The Rab7a gene in the MDA-MB-231 cells was knockout with lentivirus and the cells were divided into negative control group and Rab7a knockout group. According to the four different knockout Rab7a sequences, the Rab7a knockout group was divided into KD1 group, KD2 group, KD3 group and KD4 group. The knockout efficiencies of Rab7a gene in the MDA-MB-231 cells in various groups were detected by qPCR method, the differential genes in the highest knockout efficiency group (KD2 group) and negative control group were detected by full gene expression microarray,and the interaction between Rab7a gene and other genes was analyzed by integrated pathway analysis (IPA) software. Results: The Rab7a protein was expressed in the TNBC MDA-MB-231 cells. Compared with negative control group, the knockout efficiency of Rab7a gene in the MDA-MB-231 cells in KD2 group was the highest (P<0.01); compared with negative control group, there were 634 differential genes in the MDA-MB-231 cells in KD2 group, the number of up-regulated genes and down-regulated genes were increased(P<0.01); the differential gene analysis results showed that Rab7a knockout had the relationships with cancer, cell survival, cell cycle, cell growth,and migration. The expressions of eIF4F, IRS1 and RPS6KB1 in the Rab7a IPA network diagram were down-regulated, and the expressions of PRKAA1, IKBKE, VEGFA,and transcription factor ATF2 were up-regulated. Conclusion: Rab7a gene plays a role as a gene network in the TNBC MDA-MB-231 cells. The Rab7a gene interacts with the multiple genes and participates in the pathological processes of cancer cells.

Key words: breast neplasom, Rab7a, triple-negative breast cancer, MDA-MB-231 cells, Ingenuity Pathway Analysis

中图分类号:

R737.9

徐凤英, 刘儒, 马丽, 曲一帆, 肖伟利, 王玉珍, 谢基明. 乳腺癌MDA-MB-231细胞中Rab7a基因的网络分析[J]. 吉林大学学报(医学版), 2020, 46(03): 464-469.

XU Fengying, LIU Ru, MA Li, QU Yifan, XIAO Weili, WANG Yuzhen, XIE Jiming. Network analysis on Rab7a gene in breast cancer MDA-MB-231 cells[J]. Journal of Jilin University(Medicine Edition), 2020, 46(03): 464-469.

参考文献

[1] BOCK J B, MATERN H T, PEDEN A A, et al. A genomic perspective on membrane compartment organization[J]. Nature, 2001,409(6822):839-841.
[2] JAGER S, BUCCI C, TANIDA I, et al. Role for Rab7 in maturation of late autophagic vacuoles[J]. J Cell Sci, 2004,117(Pt 20):4837-4848.
[3] BUCCI C, THOMSEN P, NICOZIANI P, et al. Rab7:a key to lysosome biogenesis[J]. Mol Biol Cell, 2000,11(2):467-480.
[4] XIE JM, YAN Y, LIU F, et al. Knockdown of Rab7a suppresses the proliferation, migration, and xenograft tumor growth of breast cancer cells[J]. Biosci Rep, 2019,39(2):BSR20180480.
[5] COGLI L, PROGIDA C, BRAMATO R, et al. Vimentin phosphorylation and assembly are regulated by the small GTPase Rab7a[J]. Biochim Biophys Acta, 2013,1833(6):1283-1293.
[6] MARGIOTTA A, PROGIDA C, BAKKE O, et al. Rab7a regulates cell migration through Rac1 and vimentin[J]. Biochim Biophys Acta Mol Cell Res, 2017,1864(2):367-381.
[7] KOU X, YANG Y, JIANG X, et al. Vorinostat and simvastatin have synergistic effects on triple-negative breast cancer cells via abrogating Rab7 prenylation[J]. Eur J Pharmacol, 2017,813:161-171.
[8] CALVANO S E, XIAO W, RICHARDS D R, et al. A network-based analysis of systemic inflammation in humans[J]. Nature, 2005,437(7061):1032-1037.
[9] 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.
[10] ZHENG Y, ZHANG M. Disparities of breast cancer burden between China and western countries and its implication[J]. Chin J Surg, 2015,53(12):905-909.
[11] XU S, THOMPSON W, ANCOLI-ISRAEL S, et al. Cognition, quality-of-life, and symptom clusters in breast cancer:Using Bayesian networks to elucidate complex relationships[J]. Psychooncology, 2018,27(3):802-809.
[12] HARBECK N, GNANT M. Breast cancer[J]. Lancet, 2017,389(10074):1134-1150.
[13] WEIN L, LOI S. Mechanisms of resistance of chemotherapy in early-stage triple negative breast cancer (TNBC)[J]. Breast, 2017,34(Suppl 1):S27-S30.
[14] JHAN J R, ANDRECHEK E R. Triple-negative breast cancer and the potential for targeted therapy[J]. Pharmacogenomics, 2017,18(17):1595-1609.
[15] ZHAO T, DING X, YAN C, et al. Endothelial Rab7 GTPase mediates tumor growth and metastasis in lysosomal acid lipase-deficient mice[J]. J Biol Chem, 2017,292(47):19198-19208.
[16] WANG Q, LI H, ZHOU K, et al. Rab7 controls innate immunity by regulating phagocytosis and antimicrobial peptide expression in Chinese mitten crab[J]. Fish Shellfish Immunol, 2019,95:259-267.
[17] MOHAPATRA G, GAUR P, MUJAGOND P, et al. A SUMOylation-dependent switch of RAB7 governs intracellular life and pathogenesis of Salmonella typhimurium[J]. J Cell Sci, 2019,132(1):jcs222612.
[18] BARBIE T U, RITTER J L, ZHU Z, et al. Phosphorylation of RAB7 by TBK1/IKKepsilon regulates innate immune signaling in triple negative breast cancer[J]. Cancer Res, 2020,80(1):44-56.
[19] NASR Z, ROBERT F, PORCO J A J R, et al. eIF4F suppression in breast cancer affects maintenance and progression[J]. Oncogene, 2013,32(7):861-871.
[20] ADEM C, SODERBERG C L, HAFNER K, et al. ERBB2, TBX2, RPS₆KB₁, and MYC alterations in breast tissues of BRCA1 and BRCA2 mutation carriers[J]. Genes Chromosomes Cancer, 2004,41(1):1-11.

乳腺癌MDA-MB-231细胞中Rab7a基因的网络分析

Network analysis on Rab7a gene in breast cancer MDA-MB-231 cells

RichHTML

PDF (PC)

赞

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

Metrics

本文评价

推荐阅读 0

[1]	孔雯聪, 贺武斌, 苏荣健, 贾答淇, 谷艳娇, 王月, 杜晓媛. MST1R抑制剂BMS-777607对乳腺癌MCF-7细胞增殖的抑制和凋亡诱导作用[J]. 吉林大学学报(医学版), 2020, 46(03): 451-457.
[2]	申李胜男, 李思敏, 李倩, 麻帅, 韩冰. 拉曼光谱技术在乳腺癌临床应用方面的研究进展[J]. 吉林大学学报(医学版), 2020, 46(02): 413-418.
[3]	陈州华, 龚辉, 冯磊, 肖玉洁, 黄立中. LPS对乳腺癌MDA-MB-231细胞上皮间质转化的诱导作用及其对β-catenin表达的影响[J]. 吉林大学学报(医学版), 2020, 46(02): 309-315.
[4]	周春, 王光华, 张帮柱. miR-367对人乳腺癌MCF-7细胞增殖、侵袭和迁移的影响及其机制[J]. 吉林大学学报(医学版), 2019, 45(06): 1353-1360.
[5]	杨庄青, 陆眉, 杨晓娟, 王常安, 邹洁雅. 抑制TRIM24基因表达对乳腺癌MCF-7细胞增殖、凋亡、侵袭和迁移的抑制作用[J]. 吉林大学学报(医学版), 2019, 45(06): 1379-1383.
[6]	岳圆, 徐瀛濂, 王晶晶, 孙敏英, 张泽南, 赵馨越, 李宗璞, 台桂香. JNK酶活性下调对人前列腺癌、肝细胞癌和乳腺癌细胞生长的抑制作用[J]. 吉林大学学报(医学版), 2019, 45(05): 1046-1051.
[7]	于秀艳, 万广财, 孙洪帅, 朱华, 高海燕, 吴雪峰. 纤维蛋白原与乳腺癌患者预后关系的Meta分析[J]. 吉林大学学报(医学版), 2019, 45(05): 1092-1097.
[8]	谭琦, 任立群, 张祎冰, 王亚帝, 谷泽慧, 黄鹏, 陈素贤. 生物信息学在三阴性乳腺癌miRNA生物标志物筛选中的应用[J]. 吉林大学学报(医学版), 2019, 45(05): 1098-1105.
[9]	王爽, 康丽花, 关萌, 王磊, 李贝贝, 赵月, 宋艳秋, 杨婷婷. 甲磺酸阿帕替尼联合化疗在晚期乳腺癌患者治疗中的临床应用[J]. 吉林大学学报(医学版), 2019, 45(05): 1152-1158.
[10]	信瑞, 李冬, 徐慧英, 王春宇, 曲丹华, 孙海峰. miRNA-21对乳腺癌细胞放射敏感性的影响及其机制[J]. 吉林大学学报(医学版), 2019, 45(04): 772-778.
[11]	李贝贝, 杨婷婷, 关萌, 王磊, 王爽, 康丽花, 宋艳秋. HR+HER2-晚期乳腺癌内分泌治疗临床应用的研究进展[J]. 吉林大学学报(医学版), 2019, 45(03): 736-741.
[12]	周红旭, 杜烨, 于云鹤, 韩冰, 李嗣杰, 郭国强, 范志民. 乳腺癌原发灶和同侧腋窝淋巴结转移灶中激素受体及Her-2表达差异及其对预后的影响[J]. 吉林大学学报(医学版), 2019, 45(02): 376-382.
[13]	商琰红, 潘禹辰, 贾志芳, 王玥琦, 杨娜, 赵丹, 姜晶. miR200家族在乳腺癌患者血浆中表达水平的检测及其临床意义[J]. 吉林大学学报(医学版), 2019, 45(02): 383-388.
[14]	丁荣波, 张海鹏, 申李胜男, 韩冰. 乳腺癌钙化灶与其临床、病理特征和预后关系的研究进展[J]. 吉林大学学报(医学版), 2019, 45(02): 450-456.
[15]	于秀艳, 张晓伟, 丛占杰, 李铤, 吴雪峰. 乳腺癌患者血清中hMAM和CD147水平检测及其临床意义[J]. 吉林大学学报(医学版), 2018, 44(06): 1269-1274.