Effect of transcriptional coactivator with PDZ-binding motif on proliferation and migration of breast cancer cells and its mechanism
Objective To explore the effect of the transcriptional coactivator with PDZ-binding motif (TAZ) on the proliferation and migration of breast cancer cells and its molecular mechanism. Methods Human breast cancer cells MCF7 and BT549 were cultured in vitro and divided into groups A-J and a-j, respecti...
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| Format: | Article |
| Language: | zho |
| Published: |
Editorial Office of Journal of Precision Medicine
2025-06-01
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| Series: | 精准医学杂志 |
| Subjects: | |
| Online Access: | https://jpmed.qdu.edu.cn/fileup/2096-529X/PDF/1750385697275-1615101773.pdf |
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| Summary: | Objective To explore the effect of the transcriptional coactivator with PDZ-binding motif (TAZ) on the proliferation and migration of breast cancer cells and its molecular mechanism. Methods Human breast cancer cells MCF7 and BT549 were cultured in vitro and divided into groups A-J and a-j, respectively. The cells were transfected with pcDNA3.1, TAZ, PLKO.1, shTAZ, TAZ+PLKO.1, TAZ+shGLUT3, WT pGLUT3+pcDNA3.1, WT pGLUT3+TAZ, pGLUT3 mutation+pcDNA3.1, and pGLUT3 mutation+TAZ. Colony-forming assay (CFA) and Transwell assay were used to determine plating efficiency and migration rate in groups A-F and a-f. Quantitative reverse transcription PCR (RT-qPCR) was used to measure the relative expression of GLUT3 mRNA downstream of TAZ in groups A-D and a-d. The GLUT3 protein expression level in groups A-F and a-f was measured using Western blotting (WB). The binding site of TAZ transcription enhanced association domain (TEAD) and GLUT3 was predicted using the JASPER database, and the relative luciferase activity of cells in groups G-J and g-j was measured using dual luciferase reporter (DLR) assay. Results The results of the CFA showed that the plating efficiency was significantly higher in groups B/b than in groups A/a (t=23.04,25.97,P<0.05), and significantly lower in groups D/d and F/f than in groups C/c and E/e (t=9.76-42.68,P<0.05). The results of the Transwell assay showed that the cell migration rates were significantly higher in groups B/b than in groups A/a (t=30.85,29.44,P<0.05), and significantly lower in groups D/d and F/f than in groups C/c and E/e (t=12.40-33.08,P<0.05). The results of the DLR assay showed that the relative luciferase activity was significantly higher in groups H/h than in groups G/g (F=138.73,222.10,tLSD=11.09,25.81,P<0.05). The RT-qPCR results showed that the relative expression levels of GLUT3 mRNA were significantly higher in groups B/b than in groups A/a (t=15.80,14.53,P<0.05), and significantly lower in groups D/d than in groups C/c (t=4.33,4.11,P<0.05). The results of the WB showed that the relative expression levels of GLUT3 were significantly higher in groups B/b than in groups A/a (t=2.22,6.50,P<0.05), and significantly lower in groups D/d and F/f than in groups C/c and E/e (t=2.67-8.00,P<0.05). Conclusion TAZ may bind to the GLUT3 promoter region through TEAD to upregulate GLUT3 expression, thus promoting the proliferation and migration of breast cancer cells. |
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| ISSN: | 2096-529X |