Deletion of HNF1A-AS1 Suppresses the Malignant Phenotypes of Breast Cancer Cells In Vitro and In Vivo Through Targeting miRNA-20a-5p/TRIM32 Axis.

Abstract

Background: Hepatocyte nuclear factor 1 homeobox A-antisense RNA 1 (HNF1A-AS1) is a long noncoding RNA and controls human tumor development and progression. However, its expression and role in breast cancer, the most overwhelmingly occurring malignancy in women globally, remain poorly illuminated. Materials and Methods: Expression of HNF1A-AS1, miRNA (miR)-20a-5p, and tripartite motif containing 32 (TRIM32) was detected using quantitative real-time polymerase chain reaction and Western blotting. Cell proliferation, apoptosis, migration, and invasion were measured by cellTiter 96 AQueous one solution cell proliferation assay kit, flow cytometry, and transwell assays, respectively. Epithelial-mesenchymal transition (EMT) was evaluated by Western blotting, analyzing E-cadherin, N-cadherin, and vimentin expression. Mice xenograft model was generated to investigate tumor growth in vivo. The target binding among miR-20a-5p, HNF1A-AS1, and TRIM32 was confirmed by dual-luciferase reporter assay. Results: Expression of HNF1A-AS1 and TRIM32 was upregulated and miR-20a-5p was downregulated in breast cancer tumors and cell lines. Deletion of HNF1A-AS1 induced cell apoptosis rate, but suppressed cell proliferation, EMT, migration, and invasion in MDA-MB-231 and MCF-7 cells. Furthermore, HNF1A-AS1 downregulation impeded tumor growth in vivo. Interestingly, miR-20a-5p overexpression elicited the similar suppressive effects in MDA-MB-231 and MCF-7 cells, which was partially reversed by TRIM32 upregulation; besides, miR-20a-5p silencing could abolish the antitumor role of HNF1A-AS1 deletion. Notably, HNF1A-AS1 positively modulated TRIM32 expression through acting as a molecular "sponge" for miR-20a-5p. Conclusions: Knockdown of HNF1A-AS1 suppressed breast carcinogenesis presumably through targeting miR-20a-5p/TRIM32 axis, suggesting that HNF1A-AS1 might be a promising therapy target for breast cancer.

• Authors: Qingjie Meng, Linlin Wang, Yonggang Lv, Jiang Wu, Wenlong Shi
• Journal: Cancer biotherapy & radiopharmaceuticals (Cancer Biother Radiopharm) Vol. 36 Issue 1 Pg. 23-35 (Feb 2021) ISSN: 1557-8852 [Electronic] United States
• PMID: 32319789 (Publication Type: Journal Article, Observational Study)

Chemical References

• MIRN20a microRNA, human
• MicroRNAs
• RNA, Long Noncoding
• Transcription Factors
• Tripartite Motif Proteins
• long non-coding RNA HNF1A-AS1, human
• TRIM32 protein, human
• Ubiquitin-Protein Ligases

Topics

• Animals
• Breast (pathology, surgery)
• Breast Neoplasms (genetics, pathology, surgery)
• Carcinogenesis (genetics)
• Down-Regulation
• Female
• Gene Expression Regulation, Neoplastic
• Gene Knockdown Techniques
• Humans
• MCF-7 Cells
• Mastectomy
• Mice
• MicroRNAs (metabolism)
• Middle Aged
• RNA, Long Noncoding (genetics, metabolism)
• Transcription Factors (genetics)
• Tripartite Motif Proteins (genetics)
• Ubiquitin-Protein Ligases (genetics)
• Up-Regulation
• Xenograft Model Antitumor Assays