RAMP2-AS1 inhibits CXCL11 expression to suppress malignant phenotype of breast cancer by recruiting DNMT1 and DNMT3B

Li Li; Ya-Ping Gan; Hui Peng

doi:10.1016/j.yexcr.2022.113139

RAMP2-AS1 inhibits CXCL11 expression to suppress malignant phenotype of breast cancer by recruiting DNMT1 and DNMT3B

Exp Cell Res. 2022 Jul 15;416(2):113139. doi: 10.1016/j.yexcr.2022.113139. Epub 2022 Apr 4.

Authors

Li Li¹, Ya-Ping Gan², Hui Peng³

Affiliations

¹ Department of Breast Surgery, The Second Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi Province, PR China. Electronic address: 156333039@qq.com.
² Department of Gynaecology and Obstetrics, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou 510120, Guangdong Province, PR China.
³ Nanchang University, Nanchang 330006, Jiangxi Province, PR China.

PMID: 35390315
DOI: 10.1016/j.yexcr.2022.113139

Abstract

Background: Breast cancer is the most common malignancy in women populations.

Methods: RAMP2-AS1 and CXCL11 expression in breast cancer tissues and cells were determined using RT-qPCR or Western blot. RIP analysis confirmed the interaction between DNMT1, DNMT3B and RAMP2-AS1. ChIP assay verified that RAMP2-AS1 recruited DNMT1 and DNMT3B to the promoter region of CXCL11. FISH detected the sub-localization of RAMP2-AS1 in breast cancer cells. Bisulfite sequencing PCR (BSP) tested the methylation level of CXCL11. The cell viability, proliferation, migration and apoptosis were assessed by CCK-8, colony formation, transwell and flow cytometry assays, respectively. IHC was performed to evaluate the expression of Ki67, CXCL11, MMP2 in tumor tissues.

Results: The level of RAMP2-AS1 was decreased in breast cancer tissues and cells, whereas CXCL11 was highly expressed. Patients with decreased RAMP2-AS1 had a poor prognosis. RAMP2-AS1 inhibited breast cancer cell malignant phenotype. Besides, RAMP2-AS1 regulated the methylation of CXCL11 by recruiting DNMT1 and DNMT3B to the promoter region of CXCL11. RAMP2-AS1 overexpression suppressed the malignant phenotype through CXCL11 and inhibited tumor growth in vivo.

Conclusion: RAMP2-AS1 suppresses breast cancer malignant phenotype via DNMT1 and DNMT3B mediated inhibition of CXCL11.

Keywords: Breast cancer; CXCL11; DNMT1; DNMT3B; RAMP2-AS1.

MeSH terms

Breast Neoplasms* / metabolism
Cell Line, Tumor
Cell Proliferation / genetics
Chemokine CXCL11* / genetics
Chemokine CXCL11* / metabolism
DNA (Cytosine-5-)-Methyltransferase 1* / metabolism
DNA (Cytosine-5-)-Methyltransferases* / metabolism
DNA Methyltransferase 3B
Female
Gene Expression Regulation, Neoplastic
Humans
Phenotype
RNA, Long Noncoding* / genetics
Receptor Activity-Modifying Protein 2 / genetics
Receptor Activity-Modifying Protein 2 / metabolism

Substances

CXCL11 protein, human
Chemokine CXCL11
RAMP2 protein, human
RNA, Long Noncoding
Receptor Activity-Modifying Protein 2
DNA (Cytosine-5-)-Methyltransferase 1
DNA (Cytosine-5-)-Methyltransferases
DNMT1 protein, human