Hong Ji1, Meixiang Sang2, Fei Liu2, Ning Ai3, Cuizhi Geng4. 1. Department of General Surgery, the 2nd Hospital of Hebei Medical University, Shijiazhuang, Hebei, China. 2. Medical Research Center, the 4th Hospital of Hebei Medical University, Shijiazhuang, Hebei, China. 3. Department of Interventional Radiology, the 4th Hospital of Hebei Medical University, Shijiazhuang, Hebei, China. 4. Department of General Surgery, the 4th Hospital of Hebei Medical University, Shijiazhuang, Hebei, China. Electronic address: cuizhigeng@hotmail.com.
Abstract
BACKGROUND: Triple-negative breast cancer (TNBC) is highly invasive and aggressive and lacks specific molecular targets to improve the prognosis. MicroRNAs (miRNAs) serve a role in promoting and suppressing tumors in various types of malignant cancer, including TNBC. However, the regulatory mechanism of miR-124 in TNBC has still remains unclear. METHODS: Reverse transcription quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of miR-124. Cell viability was analyzed with CCK-8 assay. Cell colony formation ability was detected with colony formation assay. Cell invasion was measured with transwell assay. Dual luciferase reporter assay was conducted to verify whether ZEB2 is a target gene of miR-124. The mRNA and protein expression levels of ZEB2 and EMT markers were detected by quantitative real time PCR and western blot, respectively. RESULTS: Our results showed that miR-124 was down-regulated in TNBC tissues and cells. Overexpression of miR-124 inhibited the proliferation, metastasis and epithelial-mesenchymal transition (EMT) of TNBC cells. Furthermore, ZEB2 3'UTR was considered to be a direct target of miR-124 with luciferase reporter assay. Rescue experiments confirmed that EMT was regulated by miR-124 via suppression of ZEB2. CONCLUSION: miR-124 suppresses EMT and metastasis via ZEB2. Therefore, miR-124 may represent a potential therapeutic target for TNBC.
BACKGROUND: Triple-negative breast cancer (TNBC) is highly invasive and aggressive and lacks specific molecular targets to improve the prognosis. MicroRNAs (miRNAs) serve a role in promoting and suppressing tumors in various types of malignant cancer, including TNBC. However, the regulatory mechanism of miR-124 in TNBC has still remains unclear. METHODS: Reverse transcription quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of miR-124. Cell viability was analyzed with CCK-8 assay. Cell colony formation ability was detected with colony formation assay. Cell invasion was measured with transwell assay. Dual luciferase reporter assay was conducted to verify whether ZEB2 is a target gene of miR-124. The mRNA and protein expression levels of ZEB2 and EMT markers were detected by quantitative real time PCR and western blot, respectively. RESULTS: Our results showed that miR-124 was down-regulated in TNBC tissues and cells. Overexpression of miR-124 inhibited the proliferation, metastasis and epithelial-mesenchymal transition (EMT) of TNBC cells. Furthermore, ZEB2 3'UTR was considered to be a direct target of miR-124 with luciferase reporter assay. Rescue experiments confirmed that EMT was regulated by miR-124 via suppression of ZEB2. CONCLUSION: miR-124 suppresses EMT and metastasis via ZEB2. Therefore, miR-124 may represent a potential therapeutic target for TNBC.
Authors: Maha Siouda; Audrey D Dujardin; Laetitia Barbollat-Boutrand; Marco A Mendoza-Parra; Benjamin Gibert; Maria Ouzounova; Jebrane Bouaoud; Laurie Tonon; Marie Robert; Jean-Philippe Foy; Vincent Lavergne; Serge N Manie; Alain Viari; Alain Puisieux; Gabriel Ichim; Hinrich Gronemeyer; Pierre Saintigny; Peter Mulligan Journal: iScience Date: 2020-05-06