Function of vasohibin-2 and the mechanism of alternative splicing in triple-negative breast cancer

doi:10.3969/j.issn.1674-8115.2024.12.005

Abstract

Abstract:

Objective ·To explore the role of vasohibin-2 (VASH2) in the regulation of proliferation and metastasis of triple-negative breast cancer (TNBC) cells, and explore the mechanism of VASH2 in the occurrence and development of TNBC through regulation of gene expression and alternative splicing. Methods ·TCGA-GTEx was used to analyze the expression of VASH2 in TNBC. VASH2 methylation levels in TNBC were also analyzed. VASH2 was overexpressed in the MDA-MB-231 human TNBC cell line and transcriptome sequencing was performed. Differentially expressed genes and alternatively spliced genes regulated by VASH2 were analyzed to explore the mechanism of action of VASH2 in TNBC. Results ·VASH2 was significantly overexpressed in TNBC compared to the normal tissues. Hypomethylation of the VASH2 gene was implicated in the upregulation of VASH2 expression in TNBC. Overexpression of VASH2 caused significant differential expression of 81 genes, of which 23 genes were up-regulated and 58 genes were down-regulated. Genes with significantly altered alternative splicing levels due to VASH2 overexpressed were enriched in cell cycle and p53 signaling pathways. Conclusion ·VASH2 regulates the alternative splicing of TNBC oncogenes and promotes TNBC occurrence and development.

Key words: vasohibin-2 (VASH2), triple-negative breast cancer (TNBC), alternative splicing, p53 signaling pathway, cell cycle

CLC Number:

R737.9

WANG Wei, WANG Hongli, ALIBIYATI·i Ain, YILIYAER· Rousu, AYI NUER, YANG Liang. Function of vasohibin-2 and the mechanism of alternative splicing in triple-negative breast cancer[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2024, 44(12): 1526-1535.

Figures/Tables 6

Fig 1 Differential expression of VASH2

Fig 2 Methylated analysis of VASH2 in TNBC

Fig 3 Verification of high expression of VASH2 in MDA-MB-231 cells

Fig 4 RNA-seq analysis of VASH2-regulated transcriptome

Fig 5 RASEs in MDA-MB-231 cells and function analysis

Fig 6 Apoptosis-enriched alternative splicing genes

TrendMD

References 34

1	DERAKHSHAN F, REIS-FILHO J S. Pathogenesis of triple-negative breast cancer[J]. Annu Rev Pathol, 2022, 17: 181-204.
2	ZONG Y, PEGRAM M. Research advances and new challenges in overcoming triple-negative breast cancer[J]. Cancer Drug Resist, 2021, 4(3): 517-542.
3	MERIKHIAN P, EISAVAND M R, FARAHMAND L. Triple-negative breast cancer: understanding Wnt signaling in drug resistance[J]. Cancer Cell Int, 2021, 21(1): 419.
4	BURSTEIN H J, CURIGLIANO G, THÜRLIMANN B, et al. Customizing local and systemic therapies for women with early breast cancer: the St. Gallen International Consensus Guidelines for treatment of early breast cancer 2021[J]. Ann Oncol. 2021,32(10):1216-1235.
5	TU J J, FANG Y L, HAN D F, et al. Activation of nuclear factor-κB in the angiogenesis of glioma: insights into the associated molecular mechanisms and targeted therapies[J]. Cell Prolif, 2021, 54(2): e12929.
6	TAN X B, LIAO Z F, ZOU S Y, et al. VASH2 promotes cell proliferation and resistance to doxorubicin in non-small cell lung cancer via AKT signaling[J]. Oncol Res, 2020, 28(1): 3-11.
7	NINOMIYA Y, OZAWA S, OGUMA J, et al. Expression of vasohibin-1 and-2 predicts poor prognosis among patients with squamous cell carcinoma of the esophagus[J]. Oncol Lett, 2018, 16(4): 5265-5274.
8	SATO Y. The vasohibin family: a novel family for angiogenesis regulation[J]. J Biochem, 2013, 153(1): 5-11.
9	YAMAMOTO M, OZAWA S, NINOMIYA Y, et al. Plasma vasohibin-1 and vasohibin-2 are useful biomarkers in patients with esophageal squamous cell carcinoma[J]. Esophagus, 2020, 17(3): 289-297.
10	KITAHARA S, SUZUKI Y, MORISHIMA M, et al. Vasohibin-2 modulates tumor onset in the gastrointestinal tract by normalizing tumor angiogenesis[J]. Mol Cancer, 2014, 13: 99.
11	NORITA R, SUZUKI Y, FURUTANI Y, et al. Vasohibin-2 is required for epithelial-mesenchymal transition of ovarian cancer cells by modulating transforming growth factor-β signaling[J]. Cancer Sci, 2017, 108(3): 419-426.
12	TU M, LI Z J, LIU X, et al. Vasohibin 2 promotes epithelial-mesenchymal transition in human breast cancer via activation of transforming growth factor β 1 and hypoxia dependent repression of GATA-binding factor 3[J]. Cancer Lett, 2017, 388: 187-197.
13	MA H R, CAO L, WANG F, et al. Filamin B extensively regulates transcription and alternative splicing, and is associated with apoptosis in HeLa cells[J]. Oncol Rep, 2020, 43(5): 1536-1546.
14	YISA S B, FEI W M, YAXUN W M, et al. ATP5A1 participates in transcriptional and posttranscriptional regulation of cancer-associated genes by modulating their expression and alternative splicing profiles in HeLa cells[J]. Technol Cancer Res Treat, 2021, 20: 15330338211039126.
15	沈鑫鑫, 金磊, 赵倩, 等. 血管抑制蛋白2在乳腺癌中的表达情况及其对患者预后的影响[J]. 安徽医学, 2021, 42(2): 144-148.
	SHEN X X, JIN L, ZHAO Q, et al. Expression of vasohibin-2 and its effect on prognosis of breast cancer[J]. Anhui Medical Journal, 2021, 42(2): 144-148.
16	GAO Y, ZHANG W Z, LIU C W, et al. MiR-200 affects tamoxifen resistance in breast cancer cells through regulation of MYB[J]. Sci Rep, 2019, 9(1): 18844.
17	ZENG X, QU X J, ZHAO C Y, et al. FEN1 mediates miR-200a methylation and promotes breast cancer cell growth via MET and EGFR signaling[J]. FASEB J, 2019, 33(10): 10717-10730.
18	ANSARI J, SHACKELFORD R E, EL-OSTA H. Epigenetics in non-small cell lung cancer: from basics to therapeutics[J]. Transl Lung Cancer Res, 2016, 5(2): 155-171.
19	LI R, YANG Y E, YIN Y H, et al. Methylation and transcriptome analysis reveal lung adenocarcinoma-specific diagnostic biomarkers[J]. J Transl Med, 2019, 17(1): 324.
20	CHEN Y C, LIU X R, LI Y K, et al. Lung cancer therapy targeting histone methylation: opportunities and challenges[J]. Comput Struct Biotechnol J, 2018, 16: 211-223.
21	WANG B, YANG L, ZHAO Q, et al. Vasohibin 2 as a potential predictor of aggressive behavior of triple-negative breast cancer[J]. Am J Transl Res, 2017, 9(6): 2911-2919.
22	LI F X, HU Y J, QI S T, et al. Structural basis of tubulin detyrosination by vasohibins[J]. Nat Struct Mol Biol, 2019, 26(7): 583-591.
23	VAN DER LAAN S, LÉVÊQUE M F, MARCELLIN G, et al. Evolutionary divergence of enzymatic mechanisms for tubulin detyrosination[J]. Cell Rep, 2019, 29(12): 4159-4171.e6.
24	NIEUWENHUIS J, ADAMOPOULOS A, BLEIJERVELD O B, et al. Vasohibins encode tubulin detyrosinating activity[J]. Science, 2017, 358(6369): 1453-1456.
25	JOERGER A C, FERSHT A R. The p53 pathway: origins, inactivation in cancer, and emerging therapeutic approaches[J]. Annu Rev Biochem, 2016, 85: 375-404.
26	ZHANG H, ZHANG X, LI X, et al. Effect of CCNB1 silencing on cell cycle, senescence, and apoptosis through the p53 signaling pathway in pancreatic cancer[J]. J Cell Physiol, 2018, 234(1): 619-631.
27	MISSAOUI N, LANDOLSI H, MESTIRI S, et al. Immunohistochemical analysis of c-erbB-2, Bcl-2, p53, p21^(WAF1/Cip1), p63 and Ki-67 expression in hydatidiformmoles[J]. Pathol Res Pract,2019,215(3):446-452.
28	LO W, PARKHURST M, ROBBINS P F, et al. Immunologic recognition of a shared p53 mutated neoantigen in a patient with metastatic colorectal cancer[J]. Cancer Immunol Res,2019,7(4):534-543.
29	SCHAEFER I M, HORNICK J L, SHOLL L M, et al. Abnormal p53 and p16 staining patterns distinguish uterine leiomyosarcoma from inflammatory myofibroblastic tumour[J]. Histopathology, 2017, 70(7): 1138-1146.
30	DONNELLAN R, CHETTY R. Cyclin E in human cancers[J]. FASEB J, 1999, 13(8): 773-780.
31	KEYOMARSI K, O'LEARY N, MOLNAR G, et al. Cyclin E, a potential prognostic marker for breast cancer[J]. Cancer Res, 1994, 54(2): 380-385.
32	HUANG X, SHAO D, WU H W, et al. Genomic profiling comparison of germline BRCA and non-BRCA carriers reveals CCNE1 amplification as a risk factor for non-BRCA carriers in patients with triple-negative breast cancer[J]. Front Oncol, 2020, 10: 583314.
33	杨睿, 陈俊霞. 环状RNA hsa_circ_0058514在三阴性乳腺癌中的表达及作用研究[J]. 中国癌症杂志, 2019, 29(1): 9-18.
	YANG R, CHEN J X. Effects of circular RNA hsa_circ_0058514 on the development and progression of triple-negative breast cancer[J]. China Oncology, 2019, 29(1): 9-18.
34	LIU Z, FU Q S, WANG Y, et al. Synergy between vinorelbine and afatinib in the inhibition of non-small cell lung cancer progression by EGFR and p53 signaling pathways[J]. Biomed Pharmacother, 2021, 134: 111144.

[1]	HAN Yishan, XU Ziqi, TAO Mengyu, FAN Guangjian, YU Bo. PRMT6 promotes the proliferation and migration of breast cancer cells [J]. Journal of Shanghai Jiao Tong University (Medical Science), 2024, 44(8): 999-1010.
[2]	XUE Yu, ZHANG Hailong, LEI Ming. Expression of cancer-testis antigen SPANXB and its mechanism in affecting hepatocellular carcinoma progress [J]. Journal of Shanghai Jiao Tong University (Medical Science), 2024, 44(7): 801-813.
[3]	SHI Zelun, WANG Qing, HE Wen, FU Weijia, WANG Yingwen, HAN Xiao, ZHANG Xiaobo. Nanoplastics aggravate severe asthma by inducing DNA damage of alveolar type Ⅱ epithelial cells [J]. Journal of Shanghai Jiao Tong University (Medical Science), 2024, 44(11): 1391-1405.
[4]	WANG Xiaoling, GE Mengkai, SHEN Shaoming. PTEN-regulated alternative splicing of FoxM1 affects tumor cell migration [J]. Journal of Shanghai Jiao Tong University (Medical Science), 2023, 43(11): 1339-1347.
[5]	WANG Lanxi, MA Guanrong, JIANG Yongzhu, CHANG Xiulin, FANG Liaoqiong, BAI Jin. Effects of Escherichia coli outer membrane vesicles on proliferation of breast cancer cells and tumor growth of tumor-bearing mice [J]. Journal of Shanghai Jiao Tong University (Medical Science), 2023, 43(10): 1245-1254.
[6]	ZONG Chunyan, HE Jie, ZHANG Zhe, JIA Renbing, SHEN Jianfeng. Role of APOBEC3B in regulating replication stress of uveal melanoma [J]. Journal of Shanghai Jiao Tong University (Medical Science), 2022, 42(8): 1034-1044.
[7]	XU Feixiang, WANG Sheng, XUE Mingming, TONG Chaoyang, CHEN Yumei. Effect of altered expression of long non-coding RNA-B230352I09 on proliferation and cycle of H9C2 cardiomyocytes [J]. Journal of Shanghai Jiao Tong University (Medical Science), 2022, 42(5): 578-582.
[8]	ZHU Nan, LIU Bingya, YU Beiqin. Advances in the role of muscle blind-like protein 1 in malignant tumors [J]. Journal of Shanghai Jiao Tong University (Medical Science), 2022, 42(10): 1474-1481.
[9]	Wei HE, Yong ZUO. Effect of oridonin up-regulating PLK1 on the cell cycle of Jurkat cells [J]. JOURNAL OF SHANGHAI JIAOTONG UNIVERSITY (MEDICAL SCIENCE), 2021, 41(5): 603-611.
[10]	Qi-sheng GU, Mi-li ZHANG, Can CAO, Ji-kun LI. Association of alternative splicing and tumor immune in gastric cancer based on TCGA data set [J]. JOURNAL OF SHANGHAI JIAOTONG UNIVERSITY (MEDICAL SCIENCE), 2021, 41(4): 448-458.
[11]	CHENG Long, XU Wu-qin, ZHANG Peng, HUANG Jian-jun, LI Xiao-ning. Effect of miR-27a down-regulation on proliferation, migration and invasion in triple-negative breast cancer cells [J]. , 2020, 40(2): 188-.
[12]	GU Xiao-ping1, CHEN Meng-ping1, LIANG A-juan2, LIU Yun-xia1, SUN Hai-peng1, HUANG Ying1. Regulation of mitochondrial carrier SLC25A13 on breast cancer cell cycle in vitro [J]. , 2019, 39(8): 848-.
[13]	WANG Jia-lin, JI Di, CHEN Xiang, YANG Bo, YU Lin. Effect of miR-218-2-3P on proliferation and apoptosis of NK/T-cell lymphomatargeting SIN3A [J]. , 2019, 39(12): 1394-.
[14]	DI Fang-fang, LIU Jian-sheng, LI Shang, DU Yan-zhi . Progress on roles and mechanisms of CCAAT/enhancer binding protein β in endometrium decidualization [J]. , 2017, 37(6): 865-.
[15]	WEI Ying-qing, ZHANG Lu, HU Yu-tong, ZHANG Jian, SHEN Ying . Novel small-molecule CDK2-cyclinA2 inhibitors: design, synthesis, and biological evaluation [J]. , 2017, 37(3): 330-.