敲低核糖体发生蛋白BRIX1能够抑制三阴乳腺癌细胞的增殖、迁移和侵袭

doi:10.3969/j.issn.1009-976X.2024.01.006

摘要/Abstract

摘要： 目的探讨核糖体生物发生蛋白BRX1同源物（BRIX1）在三阴乳腺癌（TNBC）中的功能。方法通过 GEPIA2数据库分析BRIX1在正常组织和三阴乳腺癌的表达差异;设计两条 siRNA 并分别转入三阴乳腺癌细胞系 HS578T 和 MDA-MB-231 细胞内沉默 BRIX1 的表达,利用 Western Blot 验证 BRIX1 的敲低效率;使用CCK-8和集落克隆形成实验检测敲低BRIX1后细胞的增殖情况,使用Transwell实验检测敲低BRIX1后细胞的侵袭迁移能力;使用MDA-MB-231细胞,分为三组分别转入siNC、siBRIX1#1、siBRIX1#2后提取细胞的total RNA进行RNA-seq检测,进行基因表达差异分析。结果 GEPIA2数据库结果显示三阴乳腺癌中高表达BRIX1（P<0.05）,在HS578T 和 MDA-MB-231 细胞内敲低,克隆形成数目减少（P<0.05）;cck-8检测OD值减弱,证明敲低BRIX1后细胞增殖活性下降;TranswellTM实验显示敲低BRIX1后,乳腺癌侵袭迁移能力减弱;RNA-Seq数据进行GO功能富集分析生物学过程（BP）,分子功能（MF）,细胞组成（CC）,最终展示了top10相关性较高的信号通路,结果显示敲低BRIX1后,ERBB-ERK信号通路和WNT信号通路被激活,P53信号通路被抑制。结论 BRIX1基因敲低后明显抑制三阴乳腺癌细胞的增殖能力和迁移侵袭,提示BRIX1基因具有成为三阴乳腺癌治疗的潜在分子靶点。

关键词: 三阴乳腺癌, BRIX1, 细胞增殖迁移侵袭

Abstract: Objective To investigate the function of BRIX1, a large subunit component of ribosome, in triple negative breast cancer cell line in vitro. Methods The expression difference of BRIX1 in normal tissue and triple negative breast cancer was analyzed by GEPIA2 database. Two SiRNAs were designed and transferred into triple negative breast cancer cell lines HS578T and MDA-MB-231 respectively to silence the expression of BRIX1, and Western Blot was used to verify the knock inefficiency of BRIX1. The proliferation of cells after BRIX1 knockdown was detected by CCK-8 and colony cloning formation assay, and the invasion and migration ability of cells after BRIX1 knockdown was detected by Transwell assay. MDA-MB-231 cells were divided into three groups, which were transferred into siNC, siBRIX1#1 and siBRIX1#2 respectively, and total RNA of the cells was extracted for RNA-Seq detection, and the difference of gene expression was analyzed. Results The results of GEPIA2 database showed that BRIX1 was highly expressed in triple negative breast cancer （P<0.05）, and the number of clones was decreased in HS578T and MDA-MB-231 cells （P<0.05）. The OD value of cck8 was decreased, which proved that the cell proliferation activity decreased after knocking down BRIX1. TranswellTM experiment showed that the invasion and migration ability of breast cancer was weakened after knocking down BRIX1. Biological process （BP）, molecular function （MF）, and cell composition （CC） of RNA-seq data were enriched by GO, and the top10 signaling pathways with high correlation were finally revealed. The results showed that after knockdown BRIX1, ERBB-ERK and WNT signaling pathways were activated, while P53 signaling pathways were inhibited. Conclusion Down-regulated BRIX1 expression could significantly inhibited the proliferation, migration and invasion of triple negative breast cancer cells, it suggesting that BRIX1 gene could be a potential molecular target for triple negative breast cancer therapy.

Key words: triple negative breast cancer, BRIX1, proliferation, migration and invasion

中图分类号:

R739.7

董一鸣, 石川平, 廖建友. 敲低核糖体发生蛋白BRIX1能够抑制三阴乳腺癌细胞的增殖、迁移和侵袭[J]. 岭南现代临床外科, 2024, 24(01): 42-48.

DONG Yi-ming, SHI Chuan-ping, LIAO Jian-you. Effects of knockdown BRIX1 on proliferation, migration and invasion of triple negative breast cancer cells[J]. Lingnan Modern Clinics In Surgery, 2024, 24(01): 42-48.

参考文献

[1] Sung H, Ferlay J, Siegel RL, et al.Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries[J]. CA Cancer J Clin, 2021, 71(3): 209-249.
[2] Liang Y, Zhang H, Song X, Yang Q.Metastatic heterogeneity of breast cancer: Molecular mechanism and potential therapeutic targets[J]. Semin Cancer Biol, 2020, 60: 14-27.
[3] Eisenhaber F, Wechselberger C, Kreil G.The Brix domain protein family-a key to the ribosomal biogenesis pathway?[J]. Trends Biochem Sci, 2001, 26(6): 345-347.
[4] Ebersberger I, Simm S, Leisegang MS, et al.The evolution of the ribosome biogenesis pathway from a yeast perspective[J]. Nucleic Acids Res, 2014, 42(3): 1509-1523.
[5] Bogengruber E, Briza P, Doppler E, et al.Functional analysis in yeast of the Brix protein superfamily involved in the biogenesis of ribosomes[J]. FEMS Yeast Res, 2003, 3(1): 35-43.
[6] Jiang G, Teramoto Y, Goto T, et al.Identification of BXDC2 as a Key Downstream Effector of the Androgen Receptor in Modulating Cisplatin Sensitivity in Bladder Cancer[J]. Cancers, 2021, 13(5):975.
[7] Xia C, Dong X, Li H, et al.Cancer statistics in China and United States, 2022: profiles, trends, and determinants[J]. Chinese Medical Journal, 2022, 135(5): 584-590.
[8] 张希. 新辅助化疗方案治疗三阴乳腺癌的应用及临床预后分析[J]. 临床医药文献电子杂志, 2019, 6(77): 82+94.
[9] Orsolic I, Jurada D, Pullen N, et al. The relationship between the nucleolus and cancer: Current evidence and emerging paradigms [J]. Semin Cancer Biol, 2016, 37-38: 36-50.
[10] Truitt ML, Ruggero D.New frontiers in translational control of the cancer genome[J]. Nat Rev Cancer, 2016, 16(5): 288-304.
[11] Bruno PM, Liu Y, Park GY, et al.A subset of platinum-containing chemotherapeutic agents kills cells by inducing ribosome biogenesis stress[J]. Nat Med, 2017, 23(4): 461-471.
[12] Bywater MJ, Poortinga G, Sanij E, et al.Inhibition of RNA polymerase I as a therapeutic strategy to promote cancer-specific activation of p53[J]. Cancer Cell, 2012, 22(1): 51-65.
[13] Drygin D, Siddiqui-Jain A, O′Brien S, et al. Anticancer activity of CX-3543: a direct inhibitor of rRNA biogenesis[J]. Cancer Res, 2009, 69(19): 7653-7661.
[14] Cao X, Khitun A, Harold CM, et al.Nascent alt-protein chemoproteomics reveals a pre-60S assembly checkpoint inhibitor[J]. Nat Chem Biol, 2022, 18(6): 643-651.
[15] Ge J, Huang X, Wang P, Lu C.Expression of biogenesis of ribosomes BRX1 is associated with malignant progression and prognosis in colorectal cancer[J]. Transl Cancer Res, 2020, 9(9): 5595-5602.
[16] Wang L, Zhang Z, Li Y, et al.Integrated bioinformatic analysis of RNA binding proteins in hepatocellular carcinoma[J]. Aging (Albany NY), 2020, 13(2): 2480-2505.
[17] Zhang LH, Zhuo HQ, Hou JJ, et al.Proteomic signatures of infiltrative gastric cancer by proteomic and bioinformatic analysis[J]. World J Gastrointest Oncol, 2022, 14(11): 2097-2107.
[18] Jung HM, Choi SJ, Kim JK.Expression profiles of SV40-immortalization-associated genes upregulated in various human cancers[J]. J Cell Biochem, 2009, 106(4): 703-713.