CRISPR/Cas9‑mediated EZH2 knockout suppresses the proliferation and migration of triple‑negative breast cancer cells

Mao,Qiqi; Wu,Peibin; Li,Haochen; Fu,Xiaolan; Gao,Xuechen; Yang,Lei

doi:10.3892/ol.2023.13929

CRISPR/Cas9‑mediated EZH2 knockout suppresses the proliferation and migration of triple‑negative breast cancer cells

Authors:
- Qiqi Mao
- Peibin Wu
- Haochen Li
- Xiaolan Fu
- Xuechen Gao
- Lei Yang
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Affiliations: Department of General Surgery, Ningbo Medical Center Lihuili Hospital, Ningbo, Zhejiang 315000, P.R. China, Faculty of Preclinical Medicine, Guangxi University of Chinese Medicine, Nanning, Guangxi 530000, P.R. China, Department of Healthcare, Tianjin International Travel Healthcare Center (Tianjin Customs Port Outpatient Department), Tianjin 300000, P.R. China, Department of Clinical Laboratory, The First Affiliated Hospital of Guangxi University of Chinese Medicine, Nanning, Guangxi 530000, P.R. China, Department of Clinical Laboratory, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu 215000, P.R. China
Published online on: June 22, 2023 https://doi.org/10.3892/ol.2023.13929
Article Number: 343
Copyright: © Mao et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Triple‑negative breast cancer (TNBC) is an aggressive subtype of BC characterized by extensive intratumoral heterogeneity. Compared with other types of BC, TNBC is more prone to invasion and metastasis. The aim of the present study was to determine whether adenovirus‑mediated clustered regulatory interspaced short palindromic repeats (CRISPR)/Cas9 system is capable of effectively targeting enhancer of zeste homolog 2 (EZH2) in TNBC cells and lay an experimental basis for the investigation of the CRISPR/Cas9 system as a gene therapy for BC. In the present study, EZH2 was knocked out in MDA‑MB‑231 cells using the CRISPR/Cas9 gene editing tool to create EZH2‑knockout (KO) group (EZH2‑KO group). Moreover, the GFP knockout group (control group), and a blank group (Blank group), were employed. The success of vector construction and EZH2‑KO were verified by T7 endonuclease I (T7EI) restriction enzyme digestion, mRNA detection and western blotting. Changes in proliferation and migration ability of MDA‑MB‑231 cells following gene editing were detected by MTT, wound healing, Transwell and in vivo tumor biology assays. As indicated by the results of mRNA and protein detection, the mRNA and protein expression of EZH2 were significantly downregulated in the EZH2‑KO group. The difference in EZH2 mRNA and protein between the EZH2‑KO and the two control groups was statistically significant. MTT, wound healing and transwell assay suggested that the proliferation and migration ability of MDA‑MB‑231 cells in the EZH2‑KO group were significantly decreased after EZH2 knockout. In vivo, the tumor growth rate in the EZH2‑KO group was significantly lower than that in the control groups. In brief, the present study revealed that the biological functions of tumor cells were inhibited after EZH2 knockout in MDA‑MB‑231 cells. The aforementioned findings suggested that EZH2 can have a key role in the development of TNBC.

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