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Upregulation of p300 in paclitaxel-resistant TNBC: implications for cell proliferation via the PCK1/AMPK axis

The Pharmacogenomics Journal volume 24, Article number: 5 (2024) Cite this article

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Abstract

Objective

To explore the role of p300 in the context of paclitaxel (PTX) resistance in triple-negative breast cancer (TNBC) cells, focusing on its interaction with the phosphoenolpyruvate carboxykinase 1 (PCK1)/adenosine monophosphate-activated protein kinase (AMPK) pathway.

Methods

The expression of p300 and PCK1 at the messenger ribonucleic acid (mRNA) level was detected using a quantitative polymerase chain reaction. The GeneCards and GEPIA databases were used to investigate the relationship between p300 and PCK1. The MDA-MB-231/PTX cell line, known for its PTX resistance, was chosen to understand the specific role of p300 in such cells. The Lipofectamine™ 3000 reagent was used to transfer the p300 small interfering RNA and the overexpression of PCK1 plasmid into MDA-MB-231/PTX. The expression levels of p300, PCK1, 5′AMPK and phosphorylated AMPK (p-AMPK) were determined using the western blot test.

Results

In TNBC cancer tissue, the expression of p300 was increased compared with TNBC paracancerous tissue (P < 0.05). In the MDA-MB-231 cell line of TNBC, the expression of p300 was lower than in the PTX-resistant TNBC cells (MDA-MB-231/PTX) (P < 0.05). The PCK1 expression was decreased in the TNBC cancer tissue compared with TNBC paracancerous tissue, and the PCK1 expression was reduced in MDA-MB-231/PTX than in MDA-MB-231 (P < 0.05) indicating that PCK1 was involved in the resistance function. Additionally, p-AMPK was decreased in MDA-MB-231/PTX compared with MDA-MB-231 (P < 0.05). The adenosine triphosphate (ATP) level was also detected and was significantly lower in MDA-MB-231/PTX than in MDA-MB-231 (P < 0.05). Additionally, cell proliferation increased significantly in MDA-MB-231/PTX at 48 and 72 h (P < 0.05) suggesting that MDA-MB-231/PTX cells obtained the resistance function which was associated with AMPK and ATP level. When p300 was inhibited, p-AMPK and ATP levels elevated in MDA-MB-231/PTX (P < 0.05). When PCK1 was suppressed, the ATP consumption rate decreased, and cell proliferation increased (P < 0.05). However, there were no changes in p300.

Conclusions

In MDA-MB-231/PTX, p300 can inhibit p-AMPK and ATP levels by inhibiting PCK1 expression. Our findings suggest that targeting p300 could modulate the PCK1/AMPK axis, offering a potential therapeutic avenue for overcoming PTX resistance in TNBC.

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Fig. 1: Comparing expression of p300 in the TNBC tissue, TNBC paracancerous tissue, in the TNBC cell line MDA-MB-231, and in paclitaxel resistance TNBC cells MDA-MB-231/PTX.
Fig. 2: Low expression of PCK1 in TNBC and MDA-MB-231/PTX.
Fig. 3: The results for AMPK activity and ATP levels in MDA-MB-231 and MDA-MB-231/PTX.
Fig. 4: The influence on the MDA-MB-231/PTX after p300 was inhibited.
Fig. 5: The influence on the MDA-MB-231/PTX after the PCK1 overexpressed (PCK1 OVE).

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Data availability

The datasets used and analysed during the current study are available from the corresponding author upon reasonable request.

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Funding

National Natural Science Foundation of China (grant no. 82060530).

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  1. These authors contributed equally: Peng-Wei Zhao, Jia-Xian Cui.

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  1. Laboratory of Microbiology and Immunology, School of Basic Medical Science, Inner Mongolia Medical University, No.5 Xinhua Street, Huimin District, Hohhot, 010059, China

    Peng-Wei Zhao & Jia-Xian Cui

  2. Medical Oncology, Affiliated Cancer Hospital of Inner Mongolia Medical University, No. 42 Zhaowuda Road, Saihan District, Hohhot, 010020, Inner Mongolia, China

    Xiu-Mei Wang

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Peng-Wei Zhao: Performed experimental work, data collection or management, data analysis, and manuscript writing/editing. Jia-Xian Cui: Data collection or management, data analysis, manuscript writing/editing. Xiu-Mei Wang: Conceived and designed.

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Correspondence to Xiu-Mei Wang.

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Zhao, PW., Cui, JX. & Wang, XM. Upregulation of p300 in paclitaxel-resistant TNBC: implications for cell proliferation via the PCK1/AMPK axis. Pharmacogenomics J 24, 5 (2024). https://doi.org/10.1038/s41397-024-00324-3

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